Application of primary haemocyte culture of Penaeus monodon in the assessment of cytotoxicity and genotoxicity of heavy metals and pesticides

Abstract

a b s t r a c tLack of shrimp cell lines has hindered the study of pollutants which adversely affects shrimp health and its export value. In this context a primary haemocyte culture developed from Penaeus monodon was employed for assessing the cytotoxicity and genotoxicity of two heavy metal compounds, cadmium chloride and mercuric chloride and two organophosphate insecticides, malathion and monocrotophos.Using MTT assay 12 h IC 50 values calculated were 31.09 AE 16.27 mM and 5.52 AE 1.16 mM for cadmium chloride and mercuric chloride and 59.94 AE 52.30 mg l À1 and 186.76 AE 77.00 mg l À1 for malathion and monocrotophos respectively. Employing Comet assay, DNA damage inflicted by these pollutants on haemocytes were evaluated and the pollutants induced DNA damage in >60% of the cells. The study suggested that haemocyte culture could be used as a tool for quantifying cytotoxicity and genotoxicity of aquaculture drugs, management chemicals and pollutants.

Establishment of Shrimp Cell Lines: Perception and Orientation

“…The lack of success in spontaneous and induced cell line development subsequently paved the way for the attempts on developing fusion cell line [15,16], but, with little success. More recently, researchers have succeeded in viral gene expression [42], determinations of cytotoxicity and genotoxicity [41,43], and viral multiplication [26] employing primary cell culture systems developed from different species of penaeids.…”

“…To adjust osmolality, NaCl at a concentration ranging from 6 to 12 g/l [9,19,40,49] has also been added besides the balanced salt solutions [40,78]. Addition of vitamins [41,42], proline [49,56,57,82] and glutamine [28,82] has been proven to be the choice of supplements in the growth media. In addition, lactalbumin hydrolizate at a concentration of 0.1-1 g/l [2-4, 39, 48, 55-57], tryptose phosphate broth at 2.95 mg/ml [41,42] and TC Yeastolate at 1 g/l [56,57] have also been used as the source of peptides, amino acids and carbohydrates.…”

“…Addition of vitamins [41,42], proline [49,56,57,82] and glutamine [28,82] has been proven to be the choice of supplements in the growth media. In addition, lactalbumin hydrolizate at a concentration of 0.1-1 g/l [2-4, 39, 48, 55-57], tryptose phosphate broth at 2.95 mg/ml [41,42] and TC Yeastolate at 1 g/l [56,57] have also been used as the source of peptides, amino acids and carbohydrates. As the additional energy source 0.3-2 g/l glucose [40][41][42][55][56][57] and 0.55 g/l sodium pyruvate [19] have also been supplemented.…”

“…In addition, lactalbumin hydrolizate at a concentration of 0.1-1 g/l [2-4, 39, 48, 55-57], tryptose phosphate broth at 2.95 mg/ml [41,42] and TC Yeastolate at 1 g/l [56,57] have also been used as the source of peptides, amino acids and carbohydrates. As the additional energy source 0.3-2 g/l glucose [40][41][42][55][56][57] and 0.55 g/l sodium pyruvate [19] have also been supplemented. Buffering agents such as HEPS [28,48,82] and NaHCO 3 have been incorporated by many researchers [19,28,48,49].…”

Multifactorial interaction of growth factors on Penaeus monodon lymphoid cells and the impact of IGFs in DNA synthesis and metabolic activity in vitro

“…The suspension was mixed thoroughly with the medium and seeded on to 96 well plates/25 mm 2 culture flask/ dishes (Greiner Bio-One India Pvt. Ltd., Ahmedabad, India) depending on the experiments to be followed, incubated at 25°C as an open system without CO 2 in the atmosphere (Jose et al 2011(Jose et al , 2012. With the primary cell culture thus developed, growth factors were screened based on their contribution to metabolic activity using MTT assay (Jose et al 2012).…”