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Effects of standard permissible levels of Lead (Pb) for potable waters on fish innate immune response and health compared with Pb levels found in natural waterbodies

Grayson Orvile Halley


Among the list of pollutants, the heavy metals group is attracting the focus of Aquaculturist, Environmentalist and Fisherfolks alike, since they not only affect aquatic organisms but also have the potential to ultimately affect human beings. The deleterious effects of heavy metals such as lead (Pb) on aquatic ecosystems necessitate continuous monitoring of its accumulation in key species since it affords an indication of its impact on organism’s health. In natural waters, the total Pb concentrations range between 0.05 and 10.0 mg/l (Galvin 1996). While the standard recommended a limit for water for consumption range between 0.01 and 0.05 mg/L Pb. In the current study, we exposed juvenile tilapias (Oreochromis niloticus) to waterborne Pb in five concentrations 0 (control), 0.01, 0.05, 0.25 and 1.25 mg/L Pb, over six weeks period. The present study focused on the bioaccumulation of low to moderate concentrations of Pb while simultaneously assessing the effects of waterborne Pb on O. niloticus feed intake, growth performances, blood plasma  Lysozyme, Immunoglobulin M (IgM), Complement 3 (C3) and Cortisol levels. Results from the present study showed that among the Pb exposed groups a trend emerged, which indicated a high bioaccumulation rate in fish exposed to the lower (0.01, 0.05 and 0.25 mg/L) Pb concentrations. Fish exposed to 0.01 mg/l Pb showed highest accumulation rates, after 2 weeks of exposure, they were able to accumulate muscle Pb level that was equal to the concentration of Pb to which they were exposed. Meanwhile, data showed that fish exposed to 0.05 and 0.25 mg/L waterborne Pb recorded mean muscle Pb levels equal to or above the concentration of Pb to which they were exposed in the 4th week of exposure time. Furthermore, the mean muscle Pb recorded in fish after the 6th week of exposure to 0.01 mg/L waterborne Pb was 6 times above the level of the water concentration, while fish exposed to 0.05 mg/L, 0.25 mg/L and 1.25 mg/L waterborne Pb recorded mean muscle Pb after the 6th week that was 5, 3 and 2 times above the concentration of Pb in their respective water environment. Compared with the control group no significant difference in C3 activities were observed in fish exposed to 0.01 mg/L, 0.05 mg/L, and 0.25 mg/L Pb; however, as the dose of Pb increased to 1.25 mg/L significant decrease in C3 activity was observed after 4 weeks of exposure when compared with the control group. Additionally, no significant difference in IgM activities was observed in the 0.01 mg/L, 0.05 mg/L and 0.25 mg/L Pb treated groups of fish when compared with the control respectively. However, as the dose of Pb increased to 1.25 mg/L significant decrease in IgM activity was observed after 2 weeks of exposure compared with the control group. Similarly, as the dose of Pb was increased to 1.25 mg/L significant decrease in lysozyme activity was observed after 2 weeks of exposure compared to the control group. While Pb dose 0.25 mg/L after 4 weeks of exposure showed a significant decrease in lysozyme activity compared with the control. On the other hand, results on cortisol showed no significant sustained variations in activity when the respective Pb treated groups were compared with the control. The present study also showed that 1.25 mg/L waterborne Pb significantly depressed O. niloticus feed intake, weight gain, and absolute growth rate. While the mortality records showed that 75% of deaths occurred during the first two weeks of Pb exposure and the highest mortality was recorded in the group that was exposed to 1.25 mg/L Pb. The present study concluded that at low waterborne Pb concentrations fish bioaccumulate Pb faster. Also, tilapias may have become tolerant to the low Pb exposure levels over time by producing metalloproteinase and or their organs may have coped by reaching a state of homeostasis; however, further research will be needed to verify these hypotheses.


Heavy Metals; Immunity; Bioaccumulation; Hepatosomatic Index; Fulton’s Condition Factor; Oreochromis niloticus

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Abdel-Baki AS, Dkhil, MA, Al-Quraishy S. Bioaccumulation of some heavy metals in tilapia fish relevant to their concentration in water and sediment of Wadi Hanifah, Saudi Arab [J]. African Journal of Biotechnology, 2011, 10: 2541-2547.

Abdullah A, Mehana EE, Meki A. Evaluation of Pb and cadmium levels in freshwater fish farms at Qassim region, KSA [J]. Journal of Agricultural and Veterinary Sciences, 2008, 1: 59-69.

Ahmed MS, Bibi S. Uptake and bioaccumulation of waterborne Pb (Pb) in the fingerlings of a freshwater cyprinid, Catla catla [J]. The Journal of Animal & Plant Sciences, 2010, 20: 201-207.

AVMA. Guidelines on Euthanasia: Formerly Report of the AVMA Panel on Euthanasia, 2007. http://www.

Berglind R, Dave G, Sjöbeck ML. The effects of Pb on delta-aminolevulinic acid dehydratase activity, growth, hemoglobin content, and reproduction in Daphnia magna [J]. Ecotoxicol Environ Saf, 1985, 9: 29-216.

Brown M, White R, Chaille J, Russell M, Oseto C. Evaluation of three anesthetic agents for crayfish (Orconectes virilis) [J]. Shellfish Res, 1996, 15: 433-435.

Burger J, Gochfeld M. Heavy metals in commercial fish in New Jersey. Environ Res, 2005, 99: 403-412.

Cavit K, Selim S. The Immune System Drugs in Fish: Immune Function, Immunoassay, Drugs, Recent Advances in Fish Farms, 2011, 978: 307-759.

Congleton JL, La Voie WJ. Comparison of blood chemistry values for samples collected from juvenile Chinook salmon by three methods [J]. Aquat Anim Health, 2001, 13: 168–172.

Csuros M, Csuros C. Environmental sampling and analyses for metals. Lewis Publishers, ACRC Press Company, 2002.

Dalmo RA, Bogwald J. β-glucans as conductors of immune symphonies [J]. Fish and Shellfish Immunology, 2008, 25: 384–396.

Drugs Act. United Kingdom Pb in Food Regulations. Food and HMSO, 1979. London.

Dunier M, Siwicki AK. Study of the effect of pollutants on fish defense mechanisms: In vitro influence of heavy metals on the spleen and head kidney lymphocytes and macrophages activity in carp (Cyprinus carpio). GCP Project, 1993.

Ellis AE. Innate host defense mechanisms of fish against viruses and bacteria. Dev Comp Immunol, 2001, 25: 827-839.

Ewa ŁT, Ewa DK, Paweł S, Magdalena S, Włodzimierz P. Effect of long-term dietary Pb exposure on some maturation and reproductive parameters of a female Prussian carp (Carassius gibelio) [J]. Environ Sci Poll, 2014.

Fábio PA, Lourenço AS, et al., Bioaccumulation of mercury, cadmium, zinc, chromium, and Pb in muscle, liver, and spleen tissues of a large commercially valuable catfish species from Brazil [J]. Anais da Academia Brasileira de Ciências, 2016.

Fırat Ö, Cogun HY, Yüzereroğlu TA, et al., Fish Physiol Biochem, 2011, 37: 657.

Galvin RM. Occurrence of metals in water. Water SA, 1996, 22: 7–18.

Goyer RA, Clarsksom WT. Toxic effects of metals (The Basic Science of Poisons) [J]. McGraw-Hill NY, 2001, 811–867.

Grocell M, Gerdes R, Brix KV. Influence of Ca, humic acid and pH on Pb accumulation and toxicity in the fathead minnow during prolonged waterborne Pb exposure [J]. Toxicology & Pharmacology, 2006.

Hajek G, Choczewski M, Dziaman R, Kłyszejko B. Evaluation of immobilizing methods for the Chinese mitten crab (Eriocheir sinensis). Milne-Edwards, 2009, 12-18.

Hamouda EE. Pathological studies on fish experimentally intoxicated by certain heavy metals. Faculty of Veterinary Medicine, Alexandria University, Egypt, 1996.

Hasan K, Mehmet A. Effects of Waterborne Pb Exposure in Mozambique Tilapia: Oxidative Stress, Osmoregulatory Responses, and Tissue Accumulation [J]. Journal of Aquatic Animal Aealth, 2015.

Hontela A, Gagnon A, Jumarie C. Effects of Cu on plasma cortisol and cortisol secretion by adrenocortical cells of rainbow trout (Oncorhynchus mykiss). Aquat Toxicol, 2006, 78: 59–65.

Hontela A, Rasmussen JB, Audet C, et al., Arch. Environ. Contam Toxicol, 1992, 22: 278.

NIH. Guidelines for use of zebrafish.

Iwama GK, Ackerman PA. Biochemistry and molecular biology of fishes, analytical techniques for anaesthetics [J]. Elsevier Science, Amsterdam, 1994.

Jalali R, Ghafourian H, Asef Y, Davarpanah SJ, Sepehr S. Removal and recovery of Pb using nonliving biomass of marine algae [J]. Journal of Hazardous Materials, 2002, 92: 253-262.

Jerry MN. Bioaccumulation Mechanisms. Battelle Memorial Institute of Marine Environmental Sciences, 2002.

Koller LD, Kovacic S. Decreased antibody formation in mice exposed to Pb [J]. Nature, 1974, 250: 148-150.

Koller LD. Immunosuppression produced by Pb, cadmium and mercury. Am J Vet Res, 1973, 34: 1457-1458.

Lange S, Bambir SH, Dodds AW, Bowden T, Bricknell I, Espelid S, Magnadótti R. Complement component C3 transcription in Atlantic halibut (Hippoglossus hippoglossus) larvae [J]. Fish & Shellfish Immunology, 2006, 20: 285-294.

Larsson A, Haux C, Sjöbeck ML. Fish physiology and metal pollution, results and experiences from laboratory and field studies [J]. Ecotoxicol Environ Saf, 1985, 9: 81-250.

Łuszczek-TE, Drąg-Kozak E, Popek W. Pb accumulation and elimination in tissues of Prussian carp (Carassius gibelio) after long-term dietary exposure, and depuration periods [J]. Environmental Science and Pollution Research, 2013, 20: 3.

Magnadóttir B, Guðmundsdóttir BK, Lange S, Bambir SH, Steinarsson A, Oddgeirsson M, Bowden T, Bricknell I, Dalmo R, Guðmundsdóttir S. Immunostimulation of cod (Gadus morhua) larvae and juveniles [J]. Journal of Fish Diseases, 2006, 26: 147-155.

Magnadottir B, Jonsdottir H, Helgason S, Bjornsson B, Jorgensen TO, Pilstrom L. Humoral immune [J]. Veterinarni Medicina, 1999, 56: 486–503.

Magnadottir B, Lange S, Gudmundsdottir S, Bogwald J, Dalmo RA. Ontogeny of humoral immune parameters in fish [J]. Fish and Shellfish Immunology, 2005, 19: 429–439.

Magnadottir B. Innate immunity of fish [J]. Fish and Shellfish Immunology, 2006, 20: 137–151.

Marcel MP, Luis RMC, Rogelio RE. Cortisol and Glucose, reliable indicators of fish stress? [J]. Pan-American Journal of Aquatic, 2009, 4: 158-178.

Marking LL, Meyer FP. Are better anesthetics needed in fisheries? [J]. Fisheries, 1985, 10: 2–5.

Mathan R, Manoharan S, Chokkalingam K. Hormonal responses of the fish (Cyprinus carpio) to environmental Pb exposure [J]. African Journal of Biotechnology, 2009, 4154-4158.

McCoy CP, Hara TM, Bennett LW, Boyle CR, Lynn BC. Liver and kidney concentrations of zinc, copper and cadmium in channel catfish (Ictalurus punctatus) [J]. Vet Hum Toxicol, 1995, 37: 11–15.

Moore PG, Rainbow PS. Copper and zinc in an ecological series of talitrodean Amphipoda. UK. Ecologia, 1987, 73: 120-126.

Muriel D. Water pollution and immunosuppression of freshwater fish [J]. Italian Journal of Zoology, 1996, 63: 303-309.

Nouri J, Mahvi AH, Jahed GR, Babaei AA. Regional distribution pattern of groundwater heavy metals resulting from agricultural activities [J]. Environmental Geology, 2008, 55: 1337–1343.

Omaima AS, Aboud A. Impact of pollution with Pb, mercury and cadmium on the immune response of Oreochromis niloticus. New York Science Journal, 2010, 3: 9.

Omaima AS, Aboud A. Impact of pollution with Pb, mercury and cadmium on the immune response of Oreochromis niloticus [J]. New York Science Journal, 2010, 3: 9.

O'Neill JG. Effects of intraperitoneal Pb and cadmium on the humoral immune response of Salmo trutta [J]. Environ Contamin Toxicol, 1981, 27: 42-48.

Ortuño J, Esteban MA, Meseguer J. Effect of four anaesthetics on the innate immune response of gilthead seabream (Sparus aurata) [J]. Fish Shellfish Immunology, 2002, 12: 49-59.

Passow H, Rothstein A, Clarkson TW. The general pharmacology of the heavy metals [J]. Pharmacological Reviews, 1961, 13: 185-224hrs.

Rashed MN. Monitoring of environmental heavy metals in fish from Nasser Lake. Environ Int, 2001, 27: 27-33.

Ronis MJ, Badger TM, Shema SJ, Roberson PK, Shaikh F. Reproductive toxicity and growth effects in rats exposed to Pb at different periods during development. University of Arkansas for Medical Sciences, Little Ro, 1996.

Ross LG, Geddes JA. Sedation of warm water fish species in aquaculture research [J]. Aquaculture, 1979, 16: 183 186.

Ross LG, Ross B. Anaesthetic and sedative techniques for fish. Institute of Aquaculture Handbook, University of Stirling, 1984, 45-52.

Ross LG. Restraint, anaesthesia and euthanasia, veterinary practice and procedures for ornamental fish. British Small Animal Veterinary Association, London, UK, 2001.

Schreck CB. Accumulation and long-term effects of stress. The Biology of Animal Stress, 2000, 7: 147-157.

Shalaka S, Pragna P. Gonadosomatic and hepatosomatic indices of freshwater fish Oreochromis mossambicus in response to a plant nutrient [J]. World Journal of Zoology, 2013, 8: 110-118.

Sinah AK, Dasgupta P, Chakrabarty S, Bhat-Tacharya G, Bhatacharjee S. Bioaccumulation of heavy metals in different organs of some of the common edible fishes of Kharkai River, Jamshed pur Indian [J]. Journal of Environ Health, 2002, 46-102.

Spahn S, Sherry T. Arch Environ Contam Toxicol, 1999, 37: 377.

Strykowski JL, Schech JM. Effectiveness of recommended euthanasia methods in larval zebrafish (Danio rerio), 2015, 54: 81-84.

Tort L, Balasch JC, Tort L, Balasch JC, Mackenzi S. Fish immune system, a crossroads between innate and adaptive responses [J]. Inmunología, 2003, 277-286.

UNEP. Reference Methods for Marine Pollution Studies, Determination of total Hg in marine sediments and suspended solids by cold vapour AAS, 1985.

USEPA. Effects of exposure to heavy metals on selected freshwater fish, toxicity of copper, cadmium, chromium, and Pb to eggs and fry of seven fish species. Environmental Research Laboratory, Duluth, MN, 1976.

USEPA. Methods for measuring the acute toxicity of effluents and receiving water to freshwater and marine organisms, 2002.

Van-der-Oost RV, Beyer J, Vermeulen NPE. Fish bioaccumulation and biomarkers in environmental risk assessment [J]. Environmental Toxicology and Pharmacology, 2003, 13: 57-149.

Velcheva E, Tomova D, Arnaudova D, Arnaudov A. Morphological investigation on gills and liver of freshwater fish from Dam Lake [J]. Bulgarian Journal of Agriculture Sciences, 2010, 16: 364-368.

Venkatramreddy V, Vutukuru SS, Tchounwou PB. Ecotoxicology of Hexavalent Chromium in Freshwater Fish. Reviews on Environmental Health, 2009, 24hrs: 129–145.

Vutukuru SS. Heavy metals [J]. International Journal of Environmental Research and Public Health, 2005, 2: 456-462.

Watts M, Munday B, Burke C. Immune responses of teleost fish [J]. Australian Veterinary Journal, 2001, 79: 570–574.

Weis JS, Weis P. Effects of environmental pollutants on early fish development. Rev Aquat Sci, 1989, 1: 45–73.

WHO. Directives for water quality, 1986. Geneva.

WHO. Guidelines for Drinking Water Quality, 1985. Geneva.

WHO. Guidelines for drinking water quality, 1993. Geneva.

WHO. Guidelines for drinking water, 2005. Geneva.

WHO. Trace elements in human nutrition and health, 1996. Geneva.

Wilson JM, Bunte RM, Carty AJ. Evaluation of rapid cooling and tricaine methanesulfonate (MS222) as methods of euthanasia in zebrafish (Danio rerio). JAALAS, 2009, 48: 785-789.

Windholz M. Encyclopedia of chemicals, drugs and biologicals. Merck and Company, Inc, 1983, 816-818.

Winemiller KO. Feeding and reproductive biology of the curito (Hoplosternum littorale) in the Venezuelan llanos with comments on the possible functions of the enlarged male pectoral spines [J]. Environmental Biology of Fishes, 1987, 20: 219-227.

Wong PT, Cha YK, Kramar O, Bengert GA. Accumulation and depuration of tetramethyl Pb by rainbow trout. Water Res, 1981, 15: 621.

Wren CD, McCrimmon HR, Loescher BR. “Examination of bioaccumulation and biomagnification of metals in a Precambrian shield lake [J]. Water, Air, and Soil Pollution, 1983, 19: 277-291.

Yang R, Yao T, Xu B, Jiang G, Xin X. Accumulation features of organochlorine pesticides and heavy metals in fish from high mountain lakes and Lhasa River in the Tibetan Plateau [J]. Environmental International, 2007, 33: 151-156.

Yilmaz E, Akyurt I, Mutlu E. Effects of energetic diets on growth, blood chemistry, and liver pathology of African catfish (Clarias gariepinus) [J]. Israeli Journal of Aquaculture, 2007, 58: 191-197.

Zare S, Afaghi A, Heidari R, Asadpoor Y, Shiri S. Effects of Pb nitrate (PbNO3) on the glucose and cortisol hormone levels in common carp (Cyprinus carpio) [J]. Pak J Biol Sci, 2007, 10: 90-2587.

Zelikoff JT, Raymond A, Carlson E, Li Y, Beaman JR, Anderson M. Biomarkers of immunotoxicity in fish, from the lab to the ocean. Toxicol Lett, 2000, 112: 325-331.

Zelikoff JT. Biomarkers of immunotoxicity in fish and other non-mammalian sentinel species, predictive value for mammals [J]. Toxicology, 1998, 129: 63-71.

Zhang J, Huang WW, Liu MG, Cui JZ. Eco-social impact and chemical regimes of large Chinese rivers [J]. Water Resources, 1994, 28: 609-617.



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