Health risk assessment of heavy metals in Cyprinus carpio (Cyprinidae) from the upper Mekong River

The purposes of this research are to quantify the concentration of heavy metals (Zn, Cu, As, Pb, Cd, and Hg) in the water and fish tissues of common carp (Cyprinus carpio) in the upper Mekong River and to thereby elucidate the potential dietary health risks from fish consumption of local residents. Surface water and fish tissues (gill, muscle, liver, and intestine) from four representative sample areas (influence by a cascade of four dams) along the river were analyzed for heavy metal concentrations. Results revealed that the levels of heavy metals in fish were tissue-dependent. The highest Cu and As levels were found in the liver; the highest Zn and Pb levels occurred in the intestine, and the highest Hg level was found in the muscle. The total target hazard quotient (THQ) value for residents is > 1 for long-term fish consumption, and local residents are, therefore, exposed to a significant health risk. Results from the current study provide an overall understanding of the spatial and tissue distribution of heavy metals in water and fish body along the upper Mekong River under the influence of cascade dams and highlight the potential health risk of As for the local residents of long-term fish consumption.

     

Cadmium accumulation and elimination in tissues of juvenile olive flounder, Paralichthys olivaceus after sub-chronic cadmium exposure

Experiments were carried out to investigate the accumulation and elimination of cadmium (Cd) in tissues (gill, intestine, kidney, liver and muscle) of juvenile olive flounder, Paralichthys olivaceus, exposed to sub-chronic concentrations (0, 10, 50, 100 microg l(-1)) of Cd. Cd exposure resulted in an increased Cd accumulation in tissues of flounder with exposure periods and concentration, and Cd accumulation in gill and liver increased linearly with the exposure time. At 20 days of Cd exposure, the order of Cd accumulation in organs was gill > intestine > liver > kidney > muscle and after 30 days of exposure, those were intestine > gill > liver > kidney > muscle. An inverse relationship was observed between the accumulation factor (AF) and the exposure level, but AF showed an increase with exposure time. During the depuration periods, Cd concentration in the gill, intestine and liver decreased immediately following the end of the exposure periods. No significant difference was found Cd in concentration in the kidney and muscle during depuration periods. The order of Cd elimination rate in organs were decreased intestine > liver > gill during depuration periods.     

Bioaccumulation studies of organochlorinated pesticides in tissues of Cyprinus carpio

Freshwater fish Cyprinus carpio was selected for the study of bioaccumulation of organochlorinated pesticides in tissues like gills, muscle, intestine, kidney, and liver in a continuous fed system. The pesticides used were Aldrin, Dieldrin, BHC, and DDT. The bioaccumulation of Dieldrin was maximum of 85.0 microg g(-1) wet weight in liver tissue while minimum of 7.30 microg g(-1) wet weight for DDT at 30 days exposure time. Bioconcentration factor (BCF) has followed the same trend in liver tissue for Dieldrin and DDT. The rate of bioaccumulation was found to be maximum of 4.3879 microg g(-1) wet weight in liver tissue and minimum of 0.0021 microg g(-1) wet weight in gill tissue for 30 days exposure. As evidenced by the increasing values of BCF, pesticide uptake also showed increased trend with the increase in exposure time. A high correlation coefficient ranging between 0.7247 and 0.9616 between the pesticide concentration and exposure time was observed. Based on actual BCF values, log Kow were calculated and the values are well within the reported values of 6.5 indicating efficient relationship between BCF and log Kow because beyond the 6.5 the bioconcentration levels off.     

Changes in the levels of aspartate and alanine aminotransferase activities of Anabastestudineus (Bloch) exposed to disyston

In $\text{Anabas}$$\text{testudineus}$, the per cent changes noted in the aspartate aminotransferase (AAT) activities of gill, brain, intestine, liver, muscle and kidney until 6 hr of exposure to lethal concentration (10.5 mg/L disyston) were relatively much less when compared to those of sublethal concentration (4.0 mg/L disyston). But in the case of alanine aminotransferase (AIAT) activity, the tissues which showed such effect (paradoxical effect) were gill, brain and liver.     

Assessing the genotoxic potentials of arsenic in tilapia (Oreochromis mossambicus) using alkaline comet assay and micronucleus test

This experiment was conducted to study the genotoxic potentials of sodium arsenite (NaAsO₂) in freshwater fish Oreochromis mossambicus by using alkaline comet assay and micronucleus (MN) test. Fish were exposed to three different concentrations (3 ppm, 28 ppm and 56 ppm) of arsenic and gill, liver and blood tissue samples were collected after 48 h, 96 h and 192 h of exposure. Arsenic exposure induced DNA damage in all tissues examined in a concentration dependent manner. A significant (p < 0.05) increase in the comet tail DNA (%) of the exposed fish liver, gill, and blood was observed after 48 h and 96 h of exposure, but a decline in DNA damage was recorded in all the tissues at all the three concentrations studied after 192 h of exposure. Liver tissue exhibited significantly (p < 0.05) higher DNA damage at all the concentrations examined, followed by gill and blood. Higher liver tail DNA (51.38 ± 0.21%) refers that it is more prone to injury to arsenic toxicity than the gill and blood. In blood samples arsenic induced micronucleus formation in a concentration dependent manner and highest (5.8 ± 0.46%) value was recorded in 56 ppm after 96 h of exposure, whereas, it was decreased after 192 h of exposure at all the three concentrations of NaAsO₂ examined which refers to the DNA repairing ability of fish to arsenic toxicity. The results of this study depict the genotoxic potentials of arsenic to fish which in turns provide insight on advanced study in aquatic toxicology.     

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.     

New cytochrome P450 1B1, 1C1, 2Aa, 2Y3, and 2K genes from Chinese rare minnow (Gobiocypris rarus): Molecular characterization,basal expression and response of rare minnow CYP1s and CYP2s mRNA exposed to the AHR agonist benzo[a]pyrene

Cytochrome P450 (CYP450) genes play an important role in catalyzing oxidative metabolism of toxicants. Recently, CYP1 subfamily were discovered and reported in fish, however, little is known regarding the CYP2 isoforms in fish. In the present study, the cDNA fragments of CYP 1B1 and 1C1 and CYP2Aa, 2Y3, and 2K of rare minnow were cloned and exhibited a high amino acid sequence identity compared with their zebrafish orthologs. Basal expression showed CYP1C1 and CYP 2Aa expression were observed in all eight tissues analyzed (liver, gill, intestine, kidney, spleen, brain, skin, and muscle). CYP 1A, and 1B1 expression was found in all tissues except for muscle and skin. However, CYP 2Y3 was expressed in liver, spleen, intestine and muscle whereas CYP 2 K in liver, kidney and intestine. 4 and 100 μg L⁻¹ Benzo[a]pyrene (BaP) induced patterns showed that CYP 1A, 1B1 and 1C1 expression in liver, gill, and intestine was strongly up-regulated (p < 0.05). Furthermore, CYP 2Y3 was strongly induced in liver from BaP treatments (p < 0.05). The high induction on mRNA level of CYP1s and CYP 2Y3 by BaP could be associated with catalyzing detoxification and indicated that CYP2s may also be potential biomarker to screen AHR agonist. The high responsiveness of CYP1 and 2 genes suggested Chinese rare minnow is feasible to screen and assess pollution with AHR agonist.     

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.     

Metal accumulation in the smooth toadfish, Tetractenos glaber, in estuaries around Sydney, Australia

This study determined the metal levels in sediments and tissues of a common estuarine fish, Tetractenos glaber (smooth toadfish), from two metal contaminated and two reference estuaries near Sydney, Australia. Metal levels were highest in sediments and fish from contaminated estuaries. Gonads contained the highest metal levels followed by muscle, gill and liver. Metal accumulation was gender-dependant (e.g. male gonads were >20 times higher in As than females; female gills were >30 times higher than males for Pb). Cadmium, Pb and Ni levels in fish tissues reflected sediment levels, indicating sediment and/or dietary metal uptake. Levels of As, Co, Cd & Pb in gills showed similar patterns to other tissues, suggesting that metals may have been taken up by gills through contaminated water. Similar metal patterns in tissues and sediments suggest more than one uptake pathway. This study indicates that multiple factors influence metal accumulation in fish.     

A comparison of microplastic contamination in freshwater fish from natural and farmed sources

Contamination of aquatic systems mainly by urbanization and poor sanitation, deficient or lack of wastewater treatments, dumping of solid residues, and run off has led to the presence of particles, including manmade polymers, in tissues of many marine and freshwater species. In this study, the prevalence of microplastics (MPs) in freshwater fish from farmed and natural sources was investigated. Oreochromis niloticus from aquaculture farms in the Huila region in Colombia, and two local species (Prochilodus magdalenae and Pimelodus grosskopfii), naturally present in surface waters were sampled. Of the particles identified, fragments were the predominant type in the three tissue types (stomach, gill, and flesh) derived from farmed and natural fishes. MicroFT-IR spectroscopy was conducted on 208 randomly selected samples, with 22% of particles identified as MPs based on spectra with a match rate ≥ 70%. A total of 53% of identified particles corresponded to cellophane/cellulose, the most abundant particle found in all fish. Not all fish contained MPs: 44% of Oreochromis farmed fish contained MPs, while 75% of natural source fish contained MPs in any of its tissues. Overall, polyethylene terephthalate (PET), polyester (PES), and polyethylene (PE) were the prevalent MPs found in the freshwater fish. A broader variety of polymer types was observed in farmed fish. The edible flesh part of fish presented the lower prevalence of MPs compared to gill and stomach (gut), with gut displaying a higher frequency and diversity of MPs. This preliminary study suggests that the incidence and type of MPs varies in farmed verses natural fish sources as well as across different tissue types, with significantly less detected within the edible flesh tissues compared with stomach and gill tissues.

     

Exposure to sublethal concentrations of Zn(II) and Cu(II) changes biochemical parameters in Leporinus obtusidens

The aim of the present study was to assess the effect of the exposure of Leporinus obtusidens (Piava) to zinc and copper on catalase activity in the liver, delta-aminolevulinate dehidratase (delta-ALA-D) activity in liver, muscle, brain and kidney, and thiobarbituric reactive species (TBARS) in brain, muscle and liver. In addition, hematological parameters were measured in blood. The fish were exposed to 10% and 20% of the derived LC(50) values, 2.3 and 4.6 mg Zn l(-1) and 0.02 and 0.04 mg Cu l(-1), and sampled on days 30 and 45. Exposure to Zn(II) and Cu(II) decreased hematological parameters and also delta-ALA-D activity mainly in liver and kidney at all concentrations tested. Liver catalase activity increased after zinc or copper exposure at all concentrations and exposure times tested. Thiobarbituric reactive substances (TBARS) increased in the brain and liver of the fish exposed to zinc(II) for 45 days at both metal concentrations. In muscle, zinc(II) increased TBARS production at both exposure times and concentrations tested. Copper(II) exposure reduced the TBARS levels in liver at both concentrations and times tested. In brain, there was a decrease in TBARS levels only after 45 days of exposure. In muscle, this decrease was observed after 30 days of exposure at both concentrations. Although zinc and copper are required as microelements in the cells, our results showed that the sublethal concentrations of these metals can change biochemical parameters which may alter normal cellular function. These results pointed out the differential sensitivity of fish tissues to essential, but also toxic and environmentally relevant metals. The alterations of distinct biochemical parameters in fish tissues certainly contribute to the toxicity of Zn and Cu, and are of importance for an area that has been growing and has still been poorly explored in the literature.     

The intestinal parasite Pomphorhynchus laevis (Acanthocephala) from barbel as a bioindicator for metal pollution in the Danube River near Budapest, Hungary

Concentrations of As, Al, Ag, Ba, Bi, Cd, Co, Cr, Cu, Fe, Ga, Mg, Mn, Ni, Pb, Sb, Sn, Sr, Tl, V and Zn were analyzed by inductively coupled mass spectrometry (ICP-MS) in the intestinal helminth Pomphorhynchus laevis and its host Barbus barbus. The fish were caught in the Danube river downstream of the city of Budapest (Hungary). Ten out of twenty one elements analyzed were found at higher concentrations in the acanthocephalan than in different tissues (muscle, intestine, liver and kidney) of barbel. Considering the fish tissues, most of the elements were present at highest concentrations in liver, followed by kidney, intestine and muscle. Spearman correlation analyses indicate that there is competition for metals between the parasites and the host. The negative relationships between parasite number and metal levels in organs of the barbel support this hypothesis. The bioconcentration factors for Ag, As, Ba, Bi, Cu, Ga, Mn, Pb, Sr, Tl, and Zn showed that the parasites concentrated metals to a higher degree than the fish tissues. They accumulated the metals As, Cd, Cu, Fe, Ni, Pb, Sr and Zn even better than established bioindicators such as the mussel Dreissena polymorpha as revealed by data from the literature. The results presented here emphasize that acanthocephalans of fish are very useful as sentinels for metal pollution in aquatic ecosystems. Ratio of metal concentrations in the parasites and the host tissues provide additional information. Not including acanthocephalans in accumulation bioindication studies with fishes (as still customarily done) may lead to false results.     

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.     

Distribution and bioaccumulation of steroidal and phenolic endocrine disrupting chemicals in wild fish species from Dianchi Lake, China

The distribution and bioaccumulation of steroidal and phenolic endocrine disrupting chemicals (EDCs) were studied in various tissues of wild fish species from Dianchi Lake, China. In muscle tissue, 4-tert-octylphenol, 4-cumylphenol, 4-nonlyphenol and bisphenol A were detected in fish from each sampling site, with maximal concentrations of 4.6, 4.4, 18.9 and 83.5 ng/g dry weight (dw), respectively. Steroids (estrone, 17β-estradiol 17α-ethynylestradiol and estriol) were found at lower levels (<11.3 ng/g dw) and less frequently in muscle samples. The highest concentrations of steroids and phenols were found in liver, followed by those in gill and the lowest concentration was found in muscle. The field bioconcentration factors (BCFs) of phenols were calculated in fish species ranged from 18 to 97. Moreover, the measured tissue concentrations were utilized in order to estimate water concentration of steroids (4.4-18.0 ng/L). These results showed that steroidal and phenolic EDCs were likely ubiquitous contaminants in wild fish.     

The effects of fenvalerate on different tissues of freshwater fish Cirrhinus mrigala

The histopathological changes of fenvalerate on the gill, kidney, liver and intestine tissues of the Cirrhinus mrigala were determined by light microscopy. The fish were exposed to two sub-lethal concentrations of fenvalerate (1.5-3.0 ppb). The most common gill changes at all concentrations of fenvalerate were epithelial hyperplasia, epithelial necrosis, desquamation and lamellar fusion. Besides, epithelial lifting, oedema, swelling at the tips of secondary lamellae and curling of secondary lamellae were other histopathological changes. Necrosis of tubular epithelium, pycnotic nuclei in the hematopoietic tissue, hypertrophied epithelial cells of renal tubules, narrowing of the tubular lumen, expansion of space inside the Bowman's capsule and contraction of the glomerulus were observed in kidney tissues of fish. Hepatic lesions in the liver tissues of fish exposed to fenvalerate were characterized by congestion, cloudy swelling of hepatocytes and focal necrosis. Atrophy of epithelial cells, necrosis of epithelial cells, desquamation of mucosal epithelium and infiltration of lymphocytes into the lamina propria were detected in intestine tissues of fish after exposure to fenvalerate.     

Long‐term mercury accumulation in the presence of cadmium and lead in oseochromis aureus (Steindachner)

Abstract

The effects of cadmium and lead on chronic mercury accumulation were investigated in O. aureus. After 140 days’ exposure the accumulation of mercury in the liver, brain, gill filaments, intestine, caudal muscle, spleen, trunk kidney and eye was analysed. The exposure concentrations were 0.05, 0.10 and 0.20 mg/L for mercury alone. O. aureus was also exposed to mixtures of 0.05 mg/L mercury with lead (0.05 mg/L and 0.50 mg/L or cadmium (0.05 mg/L) and 0.10 mg/L mercury with 0.10 mg/L cadmium. In food fish, a knowledge of toxic metal accumulation patterns is of great importance because of their contribution to the human diet and, as fishmeal, to the diet of agricultural animals. The trunk kidney consistently accumulated higher concentrations of mercury than any of the other tissues investigated.     

Cloning of a river pufferfish (Takifugu obscurus) metallothionein cDNA and study of its induction profile in cadmium-exposed fish

We report here the full-length cDNA sequence of metallothionein (MT) gene from an anadromous river pufferfish, Takifugu obscurus (order: Tetradotiformes; family: Tetradontidae). Phylogenetic relationship analysis revealed that the identified MT has high sequence similarity with many Perciformes fish species. The tissue distribution and concentration- and time-dependent expression of MT mRNA were studied in fish exposed to cadmium. Liver showed the highest level of MT gene expression followed by other tissues (brain, gill and kidney) in response to cadmium exposure. Muscle showed a weak expression response of MT gene. Time-course study revealed highest early phase (at 6h) expression in the brain and late persistence of induction in the intestine. MT mRNA expression showed a concentration-dependent expression in all the tissues. However, induction in brain and liver occurred at much lower concentrations as compared to other tissues. Our results demonstrate that MT in T. obscurus is induced by cadmium exposure which indicates that it plays a functionally conserved function of metal detoxification.     

Biochemical effects of clomazone herbicide on piava (Leporinus obtusidens)

This study aims to verify the effects of the clomazone concentration used in rice fields on acetylcholinesterase (AChE), thiobarbituric acid reactive substances (TBARS), protein carbonyl and catalase activity in tissues of piava (Leporinus obtusidens). LC(50)-96h was 5.0 mg L(-1) and the fish were exposed to 1/10 of LC(50)-96 h: 0.5 mg L(-1) of clomazone for 96 and 192h. The same parameters were also assayed after a recovery period of 192 h in clean water. AChE activity was reduced only in the brain and heart of fish exposed for 96 h. AChE activity was decreased in the brain, muscle and heart tissues after 192 h of exposure. After 192 h of recovery period, AChE activity remained diminished in brain and muscle and showed a decrease in eye. However, after 192 h of recovery, AChE activity in heart was recovered. Fish showed increased TBARS levels in brain at all experimental periods. TBARS levels decreased in liver and muscle tissues after 192 h of exposure. The increase in muscle TBARS persisted in fish transferred to clean water. Protein carbonyl in the liver was increased in all periods studied including the recovery period. Catalase activity was reduced during all periods. The present study demonstrates the occurrence of disorders in AChE, TBARS, protein carbonyl and catalase activity in piava. The results also show changes in fish after exposure to an environmentally relevant concentration of clomazone. Most effects observed persisted after the recovery period. Thus, these parameters may be used to monitor clomazone toxicity in fish.     

Hematological, biochemical and ionoregulatory responses of Indian major carp Catla catla during chronic sublethal exposure to inorganic arsenic

In the present study the acute toxicity of arsenic trioxide in fingerlings of Catla catla an Indian major carp was evaluated with renewal bioassay method. The median lethal concentration of arsenic trioxide to the fish C. catla for 96 h was found to be 20.41 ppm (with 95% confidence limits). From this a non-lethal dose of (2.041 ppm; 1/10th of LC 50 96 value) was selected and fingerlings were exposed to 35 d and hematological, biochemical and ionoregulatory responses were studied at days 7, 14, 21, 28 and 35. Arsenic trioxide produced a significant increase in hemoglobin, hematocrit, WBC count, plasma GPT levels and reduction in RBC count, plasma sodium, chloride, potassium, glucose, protein, GOT, LDH levels as compared to the control group. Gill Na⁺/K⁺-ATPase activity was influenced by arsenic trioxide exposure. A biphasic response was noted in the value of MCH and MCV. However the MCHC level was not altered in arsenic trioxide treated fish throughout the study period. Results of the present investigation suggest that arsenic trioxide affects the hematological, biochemical and ionoregulatory parameters of fish and alterations of these parameters can be useful in environmental biomonitoring of arsenic contamination.