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Copper oxide nanoparticles and copper sulfate are established contaminants of water and soil. Metaphire posthuma is a common variety of earthworm distributed in moist soil of Indian subcontinent. Comparative toxicity of copper nanoparticles and copper sulfate were investigated with reference to selected immune associated parameters of earthworm. Total count, phagocytic response, generation of cytotoxic molecules (superoxide anion, nitric oxide), activities of enzymes like phenoloxidase, superoxide dismutase, catalase, acid phosphatase, alkaline phosphatase and total protein of coelomocytes were estimated under the exposures of 100, 500, 1000mg of copper oxide nanoparticles and copper sulfate per kg of soil for 7 and 14 d. A significant decrease in the total coelomocyte count were recorded with maximum depletion as 15.45 ± 2.2 and 12.5 ± 2 × 10 4 cells/ml under the treatment of 1000mg/kg of copper nanoparticles and copper sulfate for 14 d respectively. A significant decrease in generation of nitric oxide and activity of phenoloxidase were recorded upon exposure of both toxins for 7 and 14 d indicating possible decline in cytotoxic status of the organism. A maximum inhibition of superoxide dismutase activity was recorded as 0.083 ± 0.0039 and 0.055 ± 0.0057 unit/mg protein/minute against 1000mg/kg of copper nanoparticles and copper sulfate treatment for 14 d respectively. Activities of catalase and alkaline phosphatase were inhibited by all experimental concentrations of both toxins in the coelomocytes of earthworm. These toxins were recorded to be modifiers of the major immune associated parameters of M. posthuma. Unrestricted contamination of soil by sulfate and oxide nanoparticles of copper may lead to an undesirable shift in the innate immunological status of earthworm leading to a condition of immune compromisation and shrinkage in population density of this species in its natural habitat. This article is the first time report of immunological toxicity of nanoparticles and sulfate salt of copper in M.posthuma inhabiting the soil of India, an agriculture based country., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Waterbodies of India support a wide range of molluscs including Lamellidens marginalis , a pearl forming edible mussel of ecological significance. Report of copper oxide nanoparticle toxicity in Indian molluscs is limited in scientific literature. L. marginalis is a gill respiring filter feeder, which is toxicologically vulnerable to exposure of copper oxide nanoparticles liberated from electrical, textile and polymer industries. Experimental exposure of copper oxide nanoparticles for 7 days yielded a decrease in gill filtration rate, respiration rate, total count and phagocytic response of hemocytes, the chief immunoeffector cells of L. marginalis. Nanoparticle exposure resulted in decrease of phagocytic response of mussel hemocytes. Decrease in nitric oxide generation and phenoloxidase activity were recorded in L. marginalis exposed to 0.5, 1 and 5 mg copper oxide nanoparticles per litre of water for 7 and 14 days. Superoxide anion generation in hemocytes was increased under the exposure of copper oxide nanoparticles. Increase in superoxide anion and decrease in the activities of superoxide dismutase and catalase were indicative to oxidative stress in mussels. Copper oxide nanoparticle induced shift in filtration and respiration rate along with the hemocyte associated immune parameters were suggestive to an acute immunophysiological stress in L. marginalis. We estimated the functional performance of gill and physiological status of aquatic respiration in L. marginalis exposed to copper oxide nanoparticles. A parallel set of estimation of each parameter was carried out in L. marginalis exposed to identical copper sulphate concentrations to record and compare the ionic toxicity of copper in the same specimen. Unlabelled Image • Exposure of 0.5, 1, and 5 mg/L of CuONP/7 d yielded decrease in mussel hemocyte count. • CuONP declined filtration rate, respiration rate, phagocytosis, NO and PO activity. • Mussel exposed to CuONP for 7 and 14 d exhibited increased level of superoxide anion. • CuONP induced decrease in superoxide dismutase and catalase activity in hemocytes. • CuONP and copper sulphate exposure caused increase in activities of phosphatases. [ABSTRACT FROM AUTHOR]