Histopathological effects of copper and lithium in the ramshorn snail, Marisa cornuarietis (Gastropoda, Prosobranchia)

The aim of this study was to determine and quantify effects of copper and lithium in tissues of early juveniles of the ramshorn snail, Marisa cornuarietis. For this purpose, hatchlings of M. cornuarietis were exposed for 7 days to a range of five different sublethal concentrations of copper (5, 10, 25, 50, and 75 μg Cu²⁺ L⁻¹) and lithium (50, 100, 200, 1000, and 5000 μg Li⁺ L⁻¹). Both metals changed the tissue structure of epidermis, hepatopancreas, and gills, varying between slight and strong reactions, depending on the copper and lithium concentration. The histopathological changes included alterations in epithelial and mucous cells of the epidermis, swelling of hepatopancreatic digestive cells, alterations in the number of basophilic cells, abnormal apices of digestive cells, irregularly shaped cilia and changes in the amount of mucus in the gills. The most sensible organ in M. cornuarietis indicating Cu or Li pollution is the hepatopancreas (LOECs were 10 μg Cu²⁺ L⁻¹, or 200 μg Li⁺ L⁻¹). In epidermis, mantle and gills relevant effects occurred with higher LOECs (50 μg Cu²⁺ L⁻¹, or 1000 μg Li⁺ L⁻¹). Base on LOECs, our results indicated that histopathological endpoints are high sensitivity to copper and lithium compared to endpoints for embryonic developmental toxicity.     

Histopathological effects of chronic aqueous exposure to bis(tri-n-butyltin)oxide (TBTO) to environmentally relevant concentrations reveal thymus atrophy in European flounder (Platichthys flesus)

Although the use of tributyltin in antifouling paints has been banned, this compound is still a serious pollutant of the marine environment. This paper describes a unique study in which European flounder (Platichthys flesus) were chronically (8 months) exposed to bis(tri-n-butyltin)oxide (TBTO) in the water under controlled laboratory conditions. Residue levels in selected tissues (liver, muscle) and general health status indices were measured and the effects on several organs (gills, liver, mesonephros, ovary/testis, spleen, and gastrointestinal tract) were examined histopathologically. Additionally, morphometric analysis of the thymus was performed. The major finding is that exposure of flounder to 5 μg TBTO/l over a period of 8 months, resulting in body burdens comparable to high field levels, induced significant reduction of thymus volume, possibly affecting immunocompetence of the animals. Chronic exposure of European flounder to tributyltin is therefore likely to affect the general health status of this species in heavily polluted aquatic environments.     

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.     

Toxicity assessment and histopathological analysis of nano-ZnO against marine fish (Mugilogobius chulae) embryos

The toxicity of nano-materials has received increasing attention in recent years.Nevertheless,relatively few studies have focused on their oceanic distributions and toxicities.In this study,we assessed nano-ZnO toxicity in marine organisms using the yellowstriped goby(Mugilogobius chulae).The relative differences in nano-ZnO dissolution and dispersal in seawater and fresh water were also investigated.The effects of nano-ZnO on embryonic development,deformity,hatching,mortality,and histopathology were analyzed.In addition,the effects of the Zn~(2+) concentration on M.chulae hatching and mortality were compared.The results showed that nano-ZnO had higher solubility in seawater than in fresh water.Nano-ZnO significantly inhibited hatching.By the fifth day of exposure,the LC_(50) of nano-ZnO was 45.40 mg/L,and the mortality rate spiked.Hatching inhibition and lethality were dose-dependent over a range of1–25 mg/L nano-ZnO.Zn~(2+) inhibited hatching and increased lethality,but its effects were weaker than those of nano-ZnO at the same concentrations.Nano-ZnO also induced spinal bending,oedema,hypoplasia,and other deformities in M.chulae embryos and larvae.Histopathology revealed vacuolar degeneration,hepatocyte and enterocyte enlargement,and morphological abnormalities of the vertebrae.Therefore,nano-ZnO caused malformations in M.chulae by affecting embryonic growth and development.We conclude that nano-ZnO toxicity in seawater was significantly positively correlated with the associated Zn~(2+) concentration and sedimentary behaviour.The toxicity of nano-ZnO was cumulative and showed a critical point,beyond which embryonic and developmental toxicity in marine fish was observed.     

Monitoring pollution in River Mureş, Romania, part II: metal accumulation and histopathology in fish

As a part of an exposure and effect monitoring conducted along the river Mureş, Western Romania in 2004, the health status of two indigenous fish species, sneep (Chondrostoma nasus) and European chub (Leuciscus cephalus) was investigated upstream and downstream the city of Arad. In fish, histopathology was assessed in liver and gills, and heavy metals (cadmium, copper, lead and zinc) were analyzed in liver samples. In both fish species, histopathological reactions in the gills (epithelial lifting, focal proliferation of epithelial cells of primary and secondary lamellae and resulting fusion of secondary lamellae, hyperplasia and hypertrophy of mucous cells, focal inflammation and necrosis of epithelial cells) were most severe at the two sampling sites upstream Arad city, which were shown to be polluted by copper, cadmium, faecal coliforms and streptococci in a parallel study. At these two sites, also histopathology in the liver of L. cephalus was more prominent than at the two downstream sites. In C. nasus, symptoms in the liver (focal inflammation with lymphocytic infiltrations, macrophage aggregates and single cell necrosis) were also highly pronounced at the sampling site located directly downstream the municipal sewage treatment plant of Arad. With the exception of copper accumulation in L. cephalus caught at the most upstream sampling site, in both fish species cadmium and copper accumulation were exceptionally high and did not differ significantly between the four sampling sites.     

Determination of hypoxia and dietary copper mediated sub-lethal toxicity in carp, Cyprinus carpio, at different levels of biological organisation

Hypoxic events frequently occur in the aquatic environment in association with micro pollutants, including heavy metals. Only a few studies are however available on the uptake and biological responses of heavy metals under hypoxic conditions. To elucidate the phenomenon, mirror carp Cyprinus carpio L. (16.13-16.22 g) were exposed chronically to dietary copper (Cu; 250 and 500 mg kg dry wt.⁻¹) for 30 d under normoxic (8.25 mg O₂ L⁻¹) and hypoxic (∼3 mg O₂ L⁻¹) conditions and adopting an integrated approach, sub-lethal biomarker responses were determined at different levels of biological organisation. Level of oxidative DNA damage (as determined by modified Comet assay) showed strong significant difference following exposure to dietary Cu level under normoxic (1.6-fold) as well as under hypoxic condition at both Cu levels (2.1 and 2.5-folds respectively). Significant difference was also observed for haematological parameters (i.e. increased red and white blood cells, haematocrit value and haemoglobin concentration). Quantitative histology revealed alterations in tissues (i.e. liver and gills) for hypoxic and all dietary Cu treatment groups under both normoxic and hypoxic conditions suggesting a compensatory response to these organs (p < 0.05). The order of Cu accumulation in tissues (as determined by ICP-OES) was liver > intestine > kidney > gill. Interestingly, SGR under both normoxic and hypoxic conditions reduced with elevating Cu levels (p = 0.019). Overall, the results provide evidence for enhanced toxicological responses in fish following exposure to Cu either alone or in combination with hypoxic condition and lends support to the evolving viewpoint that many water quality guidelines should be revisited in terms of new ecotoxicological criteria.     

Growth and Histopathological Effects of Chronic Exposition of Marine Pejerrey Odontesthes argentinensis Larvae to Petroleum Water-Soluble Fraction (WSF)

The water-soluble fraction (WSF) of petroleum contains a mixture of polycyclic aromatic hydrocarbons, volatile hydrocarbons, phenols, and heterocyclic compounds, considered deleterious to aquatic biota. Marine “pejerrey” Odontesthes argentinensis (Teleostei: Atherinopsidae) has a great commercial importance in local fisheries and a high potential for aquaculture. The aim of this study was to evaluate the histopathological effects in “pejerrey” larvae exposed to different concentrations of petroleum WSF. The chronic toxicity test was conducted with newly hatched larvae exposed for 21 days to sublethal concentrations of WSF (2.5, 5, 10, and 20 % of WSF), plus one control. Survival and growth were significantly lower in the highest concentration. Several histopathological changes were found in the gills (e.g., hyperplasia, aneurisms, edema, and necrosis), kidney (e.g., nuclear alterations, decrease in the hematopoietic cells), and liver (e.g., hypertrophy, karyorrhexis, and karyopyknosis). An index of branchial lesion was proposed to standardize gill lesions to different pollutants.     

Histopathological changes in the earthworm Eisenia andrei associated with the exposure to metals and radionuclides

Earthworms were exposed for 56 d to a contaminated soil, from an abandoned uranium mine, and to the natural reference soil LUFA 2.2. Histological changes in earthworm’s body wall (epidermis, circular and longitudinal muscles) and gastrointestinal tract (chloragogenous tissue and intestinal epithelium) were assessed, after 0, 14 and 56 d of exposure. Results have shown alterations in all the studied tissues after 14 d of exposure (except for the intestinal epithelium), yet more severe effects were registered after 56 d of exposure. Herein we report histopathological alterations as a good biomarker for the evaluation of soil quality. We also demonstrate that morphological changes in the body wall and gastrointestinal tract, are important endpoints that could be added to earthworm’s standardized tests, for the evaluation of soil toxicity, as part of the risk assessment of contaminated areas.     

Histopathological and biochemical alternations of the heart induced by acute cadmium exposure in the freshwater crab Sinopotamon yangtsekiense

Cadmium (Cd) is a highly toxic element in water. Its toxicity has been attributed to oxidative stress mediated by free radicals. Here we investigated the effects of Cd on the histopathology, antioxidant enzymes and lipid peroxidation of crustacean heart. The freshwater crabs Sinopotamon yangtsekiense were exposed to different concentrations of Cd for 1, 3, 5 and 7 d. After exposure, histological abnormalities were discovered, including myocardial edema, vacuolar and vitreous degeneration, and infiltration of inflammatory cells. Additionally, alterations in nuclei, mitochondria, rough endoplasmic reticulum as well as myofibrils were observed. Meanwhile, superoxide dismutase (SOD) activity was significantly increased after Cd exposure. Catalase (CAT) activity was only increased in the group exposed to 14.50 mg L⁻¹ Cd on day 5 and decreased with increasing Cd concentration and exposure time. Glutathione peroxidase (GPx) activity was increased in groups treated with 29.00, 58.00 and 116.00 mg L⁻¹ on days 1 and 3, and decreased thereafter. Besides, malondialdehyde (MDA) levels were significantly increased after 3 d of Cd exposure at all the indicated concentrations. These results showed that acute Cd exposure led to harmful effects on the histology of crab heart, which are most likely linked to Cd-induced oxidative stress.     

Haematology, genotoxicity, enzymatic activity and histopathology as biomarkers of metal pollution in the shrew Crocidura russula

Haematological (WBC, RBC, Hgb and Hct) and genotoxicity (MNT) parameters, hepatic enzymatic activities (GST, GPx and GR), and a histopathological evaluation of liver, kidneys and gonads were assessed as general biomarkers of metal pollution in the shrew Crocidura russula inhabiting a pyrite mining area. Specimens exposed to metals presented a few significant alterations when compared with reference animals: GST activity decreased; micronuclei increased; and evident liver alterations related to metal exposure were observed. On the basis of all the parameters studied, age was an important factor that partly explained the observed variation, whereas sex was the least important factor. Significant correlations were also found between heavy metal concentrations and biomarkers evaluated, demonstrating the great influence of these metals in the metabolic alterations. To the best of our knowledge, these data constitute the first measurements of a battery of biomarkers in shrews from a mine site and are among the few available for insectivorous mammals.