Mitogenic effects of phytohemagglutinin (PHA-P) and lipopolysacharide extracted from E.coli on cultured lymphocyte of the Caspian Sea ship sturgeon (Acipenser nudiventris)

The present study was designed to obtain the most effective dose for phtytohemagglutinin (PHA-P) and lipopolysaccharide (LPS) extracted from E. coli to optimize the lymphocyte culture method, the highest mitotic index (MI) and metaphase plates number of ship sturgeon, Acipenser nudiventris. Twenty specimens of two-year old A. nudiventris weighing on average 137g and with an average length of 32cm were used in this study. Different doses (0, 40, 60 and 80kgml ^(-1)) of PHA-P, LPS and a combination of the most effective dose of both mitogenic factors {(40 kgml super(-1)) PHA-P+ (80kgml super(-1)) LPS} were added to the culture media and mitotic indices were calculated for each treatment. Results indicated that a dose of 40kgml^(-1) PHA-P and 80kgml ^(-1) LPS produced the highest MI (6.26 and 3.02 respectively). Using higher concentrations of PHA-P and LPS resulted in decreased MI, whereas at lower doses of these mitogenic factors, mitotic arrest of cultured lymphocytes was observed. Using a combination of the most effective dose of the two mentioned mitogens {(40kgml ^(-1)) PHA-P + (80kgml super(-1)) LPS} yielded a MI of about 3.84. A dose of 40kgml ^(-1) PHA-P produced the highest MI (6.26), the highest number of lymphocytes was cultured and the largest number of metaphase plate was counted