Abstract
The immunotoxic effects of cadmium (Cd) exposure in bivalves are poorly understood and whether or not stimulation of the immune system exacerbates Cd toxicity is unclear. The mussel Mytilus edulis was exposed to 20 or 50 μg/l total Cd for up to 11 days compared to no added Cd controls to assess immune and other physiological responses. Selected experiments were then repeated in the presence of a lipopolysaccharide (LPS) challenge with and without subsequent Cd exposure. Immune functions of hemocytes, hematology, hemolymph glucose and ion content, as well as superoxide dismutase (SOD) activity and organ pathology were measured. Cd accumulated mainly in digestive gland and gills and to a lesser extent in the adductor muscle. Exposure to 20 μg/l Cd alone caused a transient modulated of phagocytosis and increased neutral red retention (Kruskal–Wallis, p = 0.002). The higher Cd concentration also increased cytotoxicity, and decreased hemocyte count. Changes in hemolymph Na+, K+, and glucose were small or negligible. Histopathological examination showed tissue injuries consistent with inflammation and necrosis in the gills, digestive gland, and adductor muscle during Cd exposure alone. LPS injection alone and LPS + Cd caused an increase in the number of circulating hemocytes by the end of the experiment (Kruskal–Wallis, p = 0.01) and a transient rise in phagocytosis at day 4 (analysis of variance (ANOVA), p = 0.001). The LPS + Cd treatment also caused transient changes in neutral red retention and in the cytotoxicity of hemocytes compared to controls. Intracellular SOD activity did not change in hemocytes under any treatment. Tissue inflammation and pathology was greatly increased by the effect of Cd exposure with an LPS injection compared to either treatment alone. We conclude that immunostimulation with LPS can greatly increase Cd-related organ pathologies but does not necessarily alter the responses of hemocytes.
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References
Anderson RS, Oliver LM, Jacobs D (1992) Immunotoxicity of cadmium for the eastern oyster (Crassostrea virginica Gmelin, 1791): effects on hemocyte chemiluminescence. J Shellfish Res 11:31–35
Babich H, Borenfreund E (1992) Neutral red assay for toxicology in vitro. In: Watson RR (ed) In vitro methods of toxicology. CRC Press, Boca Raton, pp 237–251
Barham D, Trinder P (1972) An improved colour reagent for the determination of blood glucose by the oxidase system. Analyst 97:142–145
Best JH, Pflugmacher S, Wiegand C, Eddy FB, Metcalf JS, Codd GA (2002) Effects of enteric bacterial and cyanobacterial lipopolysaccharides, and of microcystin-LR, on glutathione S-transferase activities in zebra fish (Danio rerio). Aquat Toxicol 60:223–231
Brousseau P, Pellerin J, Morin Y, Cyi D, Blakley B, Boermens H, Fournier M (1999) Flow cytometry as a tool to monitor the disturbance of phagocytosis in the clam Mya arenaria hemocytes following in vitro exposure to heavy metals. Toxicology 3:145–156
Butler RA, Roesijadi G (2001) Disruption of metallothionein expression with antisense oligonucleotides abolishes protection against cadmium cytotoxicity in molluscan haemocytes. Toxicol Sci 59:101–107
Canesi L, Gallo G, Gavioli M, Pruzzo C (2002) Bacteria–hemocyte interactions and phagocytosis in marine bivalves. Microsc Res Tech 57:469–476
Cherkasov AS, Grewal S, Sokolova IM (2007) Combined effects of temperature and cadmium exposure on haemocyte apoptosis and cadmium accumulation in the eastern oyster Crassostrea virginica (Gmelin). J Thermal Biol 32:162–170
Coa A, Ramos Martinez JI, Barcia R (2004) Implication of PKA and PKC in the activation of the hemocytes of M. galloprovincialis Lmk by LPS and IL-2. Mol Immunol 41:45–52
Coles JA, Farley SR, Pipe RK (1995) Alteration of the immune response of the common marine mussel M. edulis resulting from exposure to cadmium. Dis Aquat Org 22:59–65
Da Ros L, Nasci C, Campesan G, Sartirello P, Stocco G, Menetto A (1995) Effects of linear alkylbenzene sulphonate (LAS) and cadmium in the digestive gland of mussels, Mytilus sp. Mar Environ Res 39:321–324
Fasulo S, Mauceri A, Giannetto A, Maisano M, Bianchi N, Parrino V (2008) Expression of metallothionein mRNAs by in situ hybridization in the gills of Mytilus galloprovincialis, from natural polluted environments. Aquat Toxicol 88:62–68
Faubel D, Lopes-Lima M, Freitas S, Pereira L, Andrade J, Checa A, Frank H, Matsuda T, Machado J (2008) Effects of Cd2+ on the calcium metabolism and shell mineralization of bivalve Anodonta cygea. Mar Freshw Behav Physiol 41:93–108
Federici G, Shaw BJ, Handy RD (2007) Toxicity of titanium dioxide nanoparticles to rainbow trout (Oncorhynchus mykiss): gill injury, oxidative stress, and other physiological effects. Aquat Toxicol 84:415–430
Galloway TS, Depledge MH (2001) Immunotoxicity in invertebrates: measurement and ecotoxicological relevance. Ecotoxicology 10:5–23
Galloway T, Handy R (2003) Immunotoxicity of organophosphorous pesticides. Ecotoxicology 12:345–363
Gomez-Mendikute A, Cajaraville MP (2003) Comparative effects of cadmium, copper, paraquat and benzo[a]pyrene on the actin cytoskeleton and production of reactive oxygen species (ROS) in mussel haemocytes. Toxicol In Vitro 17:539–546
Hemelraad J, Holwerda DA, Wijnne HJA, Zandee DI (1990) Effects of cadmium in freshwater clams. I. Interaction with essential elements in Anodonta cygnea. Arch Environ Contam Toxicol 19:686–690
Hernorth B (2003) The influence of temperature and dose on antibacterial peptide response against lipopolysccharide in the blue mussel, M. edulis. Fish Shellfish Immun 14:25–37
Ismail AS, Hooper LV (2005) Epithelial cells and their neighbors. IV. Bacterial contributions to intestinal epithelial barrier integrity. Am J Physiol 289:G779–G784
Jensen A, Bro-Rasmussen F (1992) Environmental cadmium in Europe. Rev Environ Contam Toxicol 125:101–181
Lowe DM, Fossato VU, Depledge MH (1995) Contaminant-induced lysosomal membrane damage in blood cells of mussels Mytilus galloprovincialis from the Venice Lagoon: an in vitro study. Mar Ecol Prog Ser 129:189–196
Mallatt J (1985) Fish gill structural changes induced by toxicants and other irritants: a statistical review. Can J Fish Aquat Sci 42:630–648
Matozzo V, Ballarin L, Pampanin DM, Marin MG (2001) Effects of copper and cadmium exposure on functional responses of hemocytes in the clam, Tapes philippinarum. Arch Environ Contam Toxicol 41:163–170
McCord JM, Fridovich I (1969) Superoxide dismutase. An enzymic function for erythrocuprein (hemocuprein). J Biol Chem 244:6049–6055
McIntosh LM, Robinson WE (1999) Cadmium turnover in the hemocytes of Mercenaria mercenaria (L.) in relation to hemocyte turnover. Comp Biochem Physiol C 123:61–66
Phillips DJH (1980) Toxicity and accumulation of cadmium in marine and estuarine biota. In: Nriagu JO (ed) Cadmium in the environment. John Wiley and Sons, New York, pp 425–569
Pipe RK, Coles JA (1995) Environmental contaminants influencing immune function in marine bivalve molluscs. Fish Shellfish Immun 5:581–595
Pipe RK, Coles JA, Farley SR (1995) Assays for measuring immune response in the mussel Mytilus edulis. In: Stolen JS, Fletcher TC, Smith SA, Zelikoff JT, Kaattari SL, Anderson RS, Söderhäll K, Weeks-Perkins BA (eds) Techniques in fish immunology. OS Publications, Fair Haven, pp 93–100
Pipe RK, Coles JA, Carissan FMM, Ramanathan K (1999) Copper induced immunomodulation in the marine mussel, Mytilus edulis. Aquat Toxicol 46:43–54
Potts WTW (1954) The inorganic composition of the blood of Mytlius edulis and Anodonta cygenea. J Exp Biol 31:376–385
Poulsen E, Riisgård HU, Møhlenberg F (1982) Accumulation of cadmium and bioenergetics in the mussel Mytilus edulis. Mar Biol 68:25–29
Raftos DA, Hutchinson A (1995) Cytotoxicity reactions in the solitary tunicate Styela plicata. Dev Comp Immunol 19:463–471
Roesijadi G, Young JS, Drum AS, Gurtisen JM (1984) Behaviour of trace metals in Mytilus edulis during a reciprocal transplant field experiment. Mar Ecol Prog Ser 18:155–170
Sant’Ana MG, Moraes R, Bernardi MM (2005) Toxicity of cadmium in Japanese quail: evaluation of body weight, hepatic and renal function, and cellular immune response. Environ Res 99:273–277
Sheir SK, Handy RD, Galloway TS (2010) Tissue injury and cellular immune responses to mercuric chloride exposure in the common mussel Mytilus edulis: modulation by lipopolysaccharide. Ecotoxicol Environ Saf (in press)
Smith KL, Galloway TS, Depledge MH (2000) Neuroendocrine biomarkers of pollution-induced stress in marine invertebrates. Sci Total Environ 262:185–190
Sokolova IM, Evans S, Hughes FM (2004) Cadmium-induced apoptosis in oyster hemocytes involves disturbance of cellular energy balance but no mitochondrial permeability transition. J Exp Biol 207:3369–3380
Stohs SJ, Bagchi D (1995) Oxidative mechanisms in the toxicity of metal ions. Free Radic Biol Med 18:321–336
Strömgren T (1982) Effect of heavy metals (Zn, Hg, Cu, Cd, Pb, Ni) on the length growth of Mytilus edulis. Mar Biol 72:69–72
Ulevitch RJ, Tobias PS (1995) Receptor-dependant mechanisms of cell stimulation by bacterial endotoxin. Annu Rev Immunol 13:437–457
Van Kolck M, Huijbregts MAJ, Veltman K, Hendriks AJ (2008) Estimating bioconcentration factors, lethal concentrations and critical body residues of metals in the mollusks Perna viridis and Mytilus edulis using ion characteristics. Environ Toxicol Chem 27:272–276
Webb M (1979) The chemistry, biochemistry and biology of cadmium. Elsevier/North-Holland Biomedical Press, Amsterdam/Oxford
Weibel ER, Kistler GS, Scherle WF (1966) Practical stereological methods for morphometric cytology. J Cell Biol 30:23–38
Wootton EC, Dyrynda EA, Ratcliffe NA (2003) Bivalve immunity: comparisons between the marine mussel (Mytilus edulis), the edible cockle (Cerastoderma edule) and the razo-shell (Ensis siliqua). Fish Shellfish Immun 15:195–210
Zelikoff JT, Bowser D, Squibb KS, Frenkel K (1995) Immunotoxicity of low level cadmium exposure in fish: an alternative animal model for immunotoxicological studies. J Toxicol Environ Health 45:235–248
Zorita I, Bilbao E, Schad A, Canico I, Soto M, Cajaraville MP (2007) Tissue- and cell-specific expression of metallothionein genes in cadmium- and copper-exposed mussels analyzed by in situ hybridization and RT-PCR. Toxicol Appl Pharm 220:186–196
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This study has been funded totally by a grant from the Egyptian Government through a PhD scholarship to S. Sheir.
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Sheir, S.K., Handy, R.D. Tissue Injury and Cellular Immune Responses to Cadmium Chloride Exposure in the Common Mussel Mytilus edulis: Modulation by Lipopolysaccharide. Arch Environ Contam Toxicol 59, 602–613 (2010). https://doi.org/10.1007/s00244-010-9502-9
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DOI: https://doi.org/10.1007/s00244-010-9502-9