Many populations of amphibians are declining on all six continents on which they occur. The reason for the declines is a direct response to the habitat destruction and pollution including heavy metals. Heavy metals represent a major environmental problem of increasing concern. They are generally found at very low concentrations. They are difficult to remove from the environment and cannot be chemically or biologically degraded. Some heavy metals like lead seem to lack biological functions and extremely toxic even at low concentrations. This study was aimed to investigate the haemotoxic and genotoxic potential of lead using blood parameters, the frequencies of micronuclei, and nuclear lesions in erythrocytes of Egyptian Toad Amietophrynus regularis as biomarkers. The results of this work revealed that Pb was potentially accumulated in liver and muscles based on dose received. Toad exposed to the selected doses of lead produced dose – dependent significant increases in the concentration of lead in the liver and muscle, confirming the ability of Amietophrynus regularis to take up and accumulate heavy metals from their ambient habitat. The results of the present investigation showed that the lead treatment inflicted a drastic reduction in the means of RBCs, haemoglobin, and haematocrit values in addition to remarkable increase in WBCs, impairing the major blood parameters in this investigation. Correlation analysis has demonstrated a negative effect of Pb accumulation on RBCs count, haemoglobin, and haematocrit. Oppositely, Pb in muscles and liver exhibited a positive effect in WBCs count. In this study, higher incidences of micronuclei (MN) and nuclear lesions (NL) were found in the blood of toad exposed to lead doses. Such frequencies were significantly elevated with the increasing lead doses. A positive correlation was demonstrated between the investigated heavy metals in tissues and the induction of micronucleated RBCs and nuclear abnormalities in Amietophrynus regularis. The results of this study confirm the usefulness of the erythrocyte MN and NL as powerful monitoring tools for detecting genotoxic agents in aquatic and terrestrial environment.
Rashad E. M. Said,
Samy A. Saber,
Alaa G. M. Osman,
Haemotoxic and Genotoxic Potential of Lead on the Egyptian Toad Amietophrynus regularis, International Journal of Ecotoxicology and Ecobiology.
Vol. 1, No. 3,
2016, pp. 94-102.
D. Roy, “Amphibians as environmental sentinels,” J. Biosci., 2002, 27(3): 187-188.
P. Daszak, and A. A. Cunningham, “Infectious disease and amphibian population declines,” J. Divers. Distrib 9: 141-150, 2003, 89: S78-S83.
U.S. EPA, “Methods for evaluating wetland condition: Using Amphibians in bioassessments of wetlands,” Office of Water, U.S. Environmental Protection Agency, Washington, 2002, DC. EPA-822-R-02-022.
M. Hirschfeld, D. C. Blackburn, T. M. Doherty-Bone, L. N. Gonwouo, S. Ghose, and M. Rödel, “Dramatic Declines of Montane Frogs in a Central African Biodiversity Hotspot,” “PLOS ONE | DOI:10.1371/journal.pone.0155129 May 5, 2016.
E. J. Houlahan, S. C. Findlay, R. B. Schmidt, H. A. Meyer, and L. S. Kuzmin, “Quantitative evidence for global amphibian population declines,” Nat., 2000, 404, 752–755.
D. B. Wake, “Declining amphibian populations,” Sci., 1991, 253 (5022): 860-860.
C. Carey, and J. C. Bryant, “Possible interrelations among environmental toxicants, amphibian development, and decline of amphibian populations,” Environ. Health. Perspect., 1995, 103(14): 13–17.
A. G. M. Osman, R. M. Al-Awadhi, Harabawy, A. S. A. and U. M. Mahmoud, “Evaluation of the use of proteinelectrophoresis of the African CatfishClarias gariepinus (Burchell, 1822) forbiomonitoring aquatic pollution,”Environ.Res. J., 2010, 4(3): 235-243.
I. J. Alinnor, and I. A. Obiji, “Assessment of trace metal composition in fish samples from Nworie River,” Pak. J. Nutr., 2010, 9, 81-85.
W. Haiyan, andA. o. Stuanes, “Heavy metal pollutionin air-water-soil-plant system of Suzhou city,” Hunan Province, china Water, Air, and Soil Poll, 147:79-107.
P. B. Tchounwou, C. G. Yedjou, A. K. Patlolla, and D. J. Sutton, “Heavy metal toxicity and the environment,” EXS. 2012;101: 133-64. doi: 10.1007/978-3-7643-8340-4_6.
J. Bleiler, D. Pillard, D. Barclift, A. Hawkins, and J. Speicher, “Development of a standardized approach for assessing potential risk to amphibians exposed to sediment and hydric soils," ENSR International Westford, MA, 2004.
HSDB U.S. Department of health and human services. National toxicology information program. National library ofmedicine. Bethesda, Maryland (CD ROM version), Micromedex, Denver, Colorado, Hazardous Substance Data Bank, 1995.
I. H. Elsokkary, M. A. Amer, and E. A. Shalaby, “Assessment of inorganic lead species and total organoalkyllead in some Egyptian agricultural soils,” Environ. Poll., 1995. 87(2): 225-233.
I. H. Elsokkary and Muller, G., “Assessment and speciation of chromium, nickel, lead and cadmium in the sediments of the river Nile, Egypt,” Sci Total Environ., 1990, 97/98: 455-463.
N. T. Basta, R. J. Ryan, L. R. Chaney, “Heavy metal and trace element chemistry in residual-treated soil, “.a review of impacts on metal bioavailability and sustainable land application,” Environ. Qual., 2005, 34 (1):49-63.
K. Lock, R. C. Janssen, “Influence of aging on copper bioavailability in soils,” Environ. Toxicol. Chem., 2001, 22(5): 1162-1166.
J. P. Bryer, “Bioaccumulation and effects of metal contaminated soil on great plains toads, Bufocognatus,” A dissertation in environmental toxicology, Ph.D. thesis Texas Tech University. 104. 2008.
W. Schmid, “The micronucleus test,”. Mutat. Res., 1975, 31(1): 9–15.
A. M. Campana, M. A. Panzeri, J. V. Moreno, and F. N Dulout, “Micronuclei induction in Rana catesbeiana tadpoles by the pyrethroid insecticide lambda–cyhalothrin,” Genet. Mo. l Biol., 2003, 26 (1): 99–103.
A. G. Heath, “Water pollution and fish physiology,” CRC Press, Inc, Boca Raton, Florida. 1995, 203-205.
M. C. A. B. Normann, J. C. F. Moreira, and V. V. Cardoso, “Micronuclei in red blood cells of armored catfish Hypostomus plecotomus exposed to potassium dichromate,” Afr. J. Biotechnol., 2008, 7(7), 893–896.
A. G. M. Osman, M. Koutb, and A. H. Sayed, “Use of hematological parameters to assess the efficiency of quince (Cydonia oblonga Miller) leaf extract in alleviation of the effect of ultraviolet–A radiation on African catfish Clarias gariepinus (Burchell,1822),” J. Photochem. Photobiol., 2010, B (99): 1-8.
B. O. Mgbenka, N. S. Oluah,and I. Umeike, “Effect of gammalin 20 (lindane) on differential white blood cell counts of the African catfish, Clarias albopunctatus,” Environ. Contam. Toxicol., 2003, 71(2): 248-254.
M. A. Arrieta, S. I. Peri, C. Apartin, C. E. Rosenberg, N. E. Fink, and A. Salibian, “Blood lead concentration and delta-aminolevulinic acid dehydratase activity in adult Bufo arenarum,” Arch. Physiol. Biochem., 2000. 108(3), 275-280.
M. A. Arrieta, C. E. Rosenberg, N. E. Fink, and A. Salibián, Toxicidad aguda del plomo para Bufo arenarum a dos temperaturas. In: X Congreso Argentino de Toxicología, La Plata (Argentina), September 27–29, 1999. Buenos Aires, Asociación Toxicológica Argentina, Abstr 56.
M. A. Arrieta, S. I. Peri,C. Apartin,C. E. Rosenberg, N. E. Fink,and A. Salibian, “Blood lead concentration and delta-aminolevulinic acid dehydratase activity in adult Bufo arenarum,” Arch. Physiol. Biochem., 2000, 108(3):275-280.
W. McDaniel, “Sample preparation procedure for petrochemical determination of total recoverable elements in biological tissues in "revision 1.0 Environmental monitoring systems laboratory. U.S Environmental Protection Agency,” 1991, 23–29.
A. T. AbdAllah, “Ecological and biological studies on the freshwater snail Biomphalaria glabrata the intermediate host of Schistosoma mansoni,” PhD Thesis, Al–Azhar University, Cairo, Egypt, 1996.
S. K. Arserim, and A. Mermer,“Hematology of the Uludağ Frog, Rana macrocnemis Boulenger, 1885,” in Uludağ National Park (Bursa, Turkey). Fisheries & Aquatic Sciences, 2008, 25(1): 39–46.
B. Bosch, F. Mañas, N. Gorla, and D. Aiassa, “Micronucleus test in post metamorphic Odontophrynus cordobae and Rhinella arenarum (Amphibia: Anura) for environmental monitoring,” J. Toxicol. Environ. Health Sci., 2011, 3(6): 155-163.
S. L. Shah, and A. Altindag, “Haematological parameters of tench (Tincatinca L) after acute and chronic exposure to lethal and sublethal mercury treatments,” Bull.Enviro.Cont.Toxicol., 2004, 73(5):911-918.
S. L. Shah, and A. Altindag, “Alterations in the immunological parameters of tench (Tinca tinca, L.) after acute and chronic exposure to lethal and sublethal treatments with mercury, cadmium and lead,” Turk. J. Vet. Anim. Sci., 2005, 29: 1163-1168.
T. Satake, A. Nuti-Sobrinho, O. V. Paula-Lopes, R. A. Lopes, and H. S. Leme Dos Santos, “Haematological study of Brazilian fish. III. Blood parameters in Armored Catfish Hypostomus paulinus,” I. ArsVeterinaria, Faculdade de Ciências Agrárias e Veterinárias “Campus” de Jaboticabal Unesp, 1986, 2: 179-183.
P. Reddy, and M. Bashamohideen, “Fenvalarate and cypermethrin induced changes in the haematological parameters of Cyprinus carpio,” Acta Hydroch.Hydrob., 1989, 17:101-107.
C. A. Ilizaliturri-Hernández, D. J. González-Mille, J. Mejía-Saavedra, G. Espinosa-Reyes, A. Torres-Dosal, and I. Pérez-Maldonado, “Blood lead levels, δ-ALAD inhibition, and haemoglobin content in blood of giant toad (Rhinella marina) to asses lead exposure in three areas surrounding an industrial complex in Coatzacoalcos, Veracruz, Mexico,” Database: CrossRef. Environ. Monit. Assess., 2012.
J. S. Abreu, C. M. Marzocchi-Machado, A. C. Urbaczek, L. M. Fonseca, and E. C. Urbinati,“Leukocytes respiratory burst and lysozyme level in pacu (Piaractus mesopotamicus Holmberg, 1887),” Braz. J. Biol., 2009, 69(4), 1133-1139.
S. Feng, Z. Kong, X. Wang,, L. Zhao and P. Peng, “Acute toxicity and genotoxicity of two novel pesticides on amphibian, Rana N. Hallowell,” Chemosphere, 2004,56(5):457–463.
U. Güner, and F. D. G. Muranli, “Micronucleus test, nuclear abnormalities and accumulation of Cu and Cd on Gambusia affinisBaird & Girard, 1853),” Turk. J. Fish. Aquat. Sci., 2011, 11(4): 615-622.
A. G. M. Osman, A. M. Abd El Reheem, A. M Moustafa, W. Kloas, U.M. Mahmoud, K. Y. Abuel-Fadl, “ In situ evaluation of the genotoxic potential of the river Nile: I. Micronucleus and nuclear lesion tests of erythrocytes of Oreochromis niloticus niloticus (Linnaeus, 1758) and Clarias gariepinus (Burchell, 1822), Toxicol. & Environ.Chem.,93, 5, 2011.
K. R. Carrasco, K. L. Tilbury, and M. S. Mayers, “Assessment of the piscine micronuclei test asan in situ biological indicator of chemical contaminants effects,”: doi: 10.1139/f90-237. Can. J. Fish. Aquat. Sci., 1990, 47, 2123–2136.
J. O. Nriagu, “A global assessment of natural sources of atmospheric trace metals,” Nat.,1989, 338, 47–49.
J. Pacyna,and E. Pacyna, "An assessment of global and regional emissions of trace metals to the atmosphere from anthropogenic sources worldwide," Environmental Reviews, 2001, 9:269–298.
H. Arslan, and A. M. Gizir, Heavy-metal content of roadside soil in Mersin, Turkey. Fresen. Environ. Bull., 2006, 15(1):15-20.
J. E. Fergusson, R. W. Hayes, T. S. Young, and S. H. Thief, “Heavy metal pollution by traffic in Christchurch, New Zealand, lead andcadmium content of dust, soil and plant samples,” New Zealand J. Sci., 1980,23, 293-310.
C. J. McGeer, V. K. Brix, M. James, M. J. Skeaff, K. D. Defforest, I. S. Brigham, J. W. Adams, and A. Green, “Inverse relationship between bioconcentration factor and exposure concentration for metals: implications for hazard assessment of metals in the aquatic environment,”Environ. Toxicol. Chem., 2003, 22(5): 1017–1037.
L. I. Ezemonye,and A. A. Enuneku, “Biochemical changes in the toad Bufo maculatus treated with sublethal concentrations of cadmium,” World J. Biol.Res., 2011a,1(4): 15-20.
L. I. Ezemonye, and A. A. Enuneku, “Haematological changes in Bufo maculatus treated with sublethal concentrations of cadmium,” Xenobiotics, 2011b, 1(7): 35-37.
L. I. Ezemonye,and A. A. Enuneku, “Histopathological alterations in the liver and lungs of Hoplobatrachus occipitalis exposed to sub lethal concentrations of cadmium,” Aust. j. Basic Appl. Sci., 2011c, 5(11): 1062-1068.
L. I. Ezemonye, and A. A. Enuneku, “Bioaccumulation and histopathological alterations in the flat backed toad, Bufo maculatus exposed tosub lethal concentrations of lead,” New York Sci. J., 2012a, 5(2):147-156.
L. I. Ezemonye,and A. A. Enuneku, “Hepatic bioaccumulation of cadmium in the crowned bullfrog, Hoplobatrachus occipitalis and flat backed toad, Bufo maculates,” Int. J. Aqua. Sci., 2012b, 3(1).
R. James, K. Sampath, and S. Alagrathinam, “Effects of lead on respiratory enzyme activity, glycogen and blood sugar levels of the teleost Oreochromis mossambicus (Peters) during accumulation and depuration Asian Fish. Sci., 1996, 9(2): 87-100.
D. W. Sparling, G. Linder, and C. A. Bishop, “Ecotoxicology of amphibians and reptiles (2nded.). Pensacola: Society of environmental toxicology and chemistry (SETAC)., 2010.
M. A. Arrieta, L. Bruzzone, C. Rosenberg, C. E. Apartín, C. E. Rosenberg, and A. Salibián, “Biosensors of inorganic lead exposure and effect in an adult amphibian,” Arch. Environ.Cont. Toxicol., 2004, 46 (2):224-230.
S. I. Perí, M. A. Arrieta, N. E. Fink, and A. Salibián, Delta-aminolevulinic acid dehydratase (ALAD) activity in blood of Bufo arenarum (Anura). Biol.Re., 1998, 31(4), 339-342.
C. E. Rosenberg, I. Perí, S. M. A. Arrieta, N. E.. Fink, and A. Salibián, “Red blood cell osmotic fragility in Bufo arenarum exposed to lead” Arch.Physiol.Bioch., 1998, 106(1): 19–24.
C. E. Rosenberg,N. E. Fink, M. A. Arrieta, and A. Salibia´na, “Effect of lead acetate on the in vitro engulfment and killing capability of toad (Bufo arenarum) neutrophils,” Comp. Biochem. PhysioH.. Part C, (2003): 136, 225–233.
Y. Zhang, D. Huang, D. Zhao, J. Long, G. Song, and L. An’na, “Long-term toxicity effects of cadmium and lead on Bufo raddei tadpoles,” Bull.Environ.Cont.Toxico., 2007,79 (2): 178–183.
R. Maheswaran, A. Devapaul, S. Muralidharan, B. Velmurugan, and S. Ignacimuthu, Haematological studies of fresh water fish, Clarias batrachus (L.) exposed to mercuric chloride,” Int. J.Int. Biol., 2008, 2 (1): 49-54.
J. D. Saroch, H. Nisar,, R. Shrivastav T. A. Qureshi, and S. Manohar, “Haematological studies of mercuric chloride affected freshwater catfish Clarias gariepinus fed with Spirulina. Chemical,” Biol. Physiol.Sci., 2012, 2(4) 1862-1869.
P. Allen, “Changes in the hematological profile of the cichilid, Oreochromis aureus (Steindachner (during acute inorganic mercury intoxication,” Comp. Biochem. Physiol., 1994, 108(1):17-121.
T. S. Gill, and J. C. Pant, “Erythrocytic and leukocytic responses to cadmium poisoning in fresh water fish,” Puntius conchonius Ham. Environ.Res., 1985, 36(2): 327–337.
S. Dhanekar, and S. Srivastava, “Studies on toxic effects of least effective concentration of mercury in fish: a haematological study,” Matsya, 1985, 11:75-78.
C. A. Oliveira, F. Ribeiro, and F. Neto, “Hematological findings in neotropical fish Hopilas malabaricus exposed to subchronic and dietary doses of methyl mercury, inorganic lead, and tributyltin chloride,” Envir.Res., 2006, 101(1): 74-80.
D. Singh, K. Nath, S. P. Trivedi, and Y. K. Sharma, “Impact of copper on haematological profile of freshwater fish, Channa punctatus,” Environ.Biol., 2008, 29(2): 253-257.
V. Kašuba, R. Rozgaj, A. Fuči,V. M. Varnai, and M. Piasek, “Lead acetate genotoxicity in suckling rats,” Biologia, Bratislava, 2004,59(6), 779-785.
A. G. M. Osman, A. S. and Harabawy, “Hematotoxic and Genotoxic potential of ultraviolet–A radiation on the African catfish Clarias gariepinus (Burchell, 1822),” 2010, J.Fish. Internat.5 (3): 44-53.
C. Bolognesi, E. Perrone, P. Roggieri,. D. M. Pampanin, and A. Sciutto, “Assessment of micronuclei induction in peripheral erythrocytes of fish exposed to xenobiotics under controlled conditions,” Aqu. Toxicol., 2006.,Vol. 78S. P. S93–S98.
S. Ergene, T. Cavaş, A. Celik, N. Koleli, F. Kaya, andA. Karahan, “Monitoring of nuclear abnormalities in peripheral erythrocytes of three fi sh species from the Goksu Delta (Turkey): genotoxic damage in relation to water pollution, Ecotoxicol” 2007, Vol. 16. P. 385–391.
N. Ercal, H. Gurer-Orhan, and N. Aykin-Burns, “Toxic metals and oxidative stress part I: mechanisms involved inmetal-induced oxidative damage,” Current Topics in Medicinal Chemistry, 2001, 1(6), 529-539.