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Siwicki A. K., J. Fuller Jr., S. Nissen, M. Morand, F. Pozet, F. Vincent, B. Kazuƒ: Effect of HMB (β-hydroxy-β− methylbutyrate) on in vitro Proliferative Responses of Sheatfish (Silurus glanis) and Catfish (Ictalurus melas) Lymphocytes Stimulated by Mitogens. Acta Vet. Brno 2004, 73: 119-122. The aim of this work was to study the in vitro effect of β-hydroxy-β-methylbutyrate (HMB) on the proliferative response of sheatfish ( Silurus glanis) and catfish (Ictalurus melas) lymphocytes stimulated by mitogens at temperatures of 20 °C and 30 °C. The pronephros lymphocytes were isolated from 20 sheatfish and 20 catfish, respectively. Mitogens, ConA at concentration of 64 µg/ml or LPS at concentration of 160 µg/ml were used for stimulation of lymphocytes. The plates were incubated for 72 h at 20 °C and 30 °C without CO 2. The MTT method was used for the study of proliferative responses of lymphocytes stimulated by mitogens. The plates were read on a microreader, using the test wavelength of 620 nm. Net optical density (OD) values were obtained by subtracting the mean OD of negative control cells (not stimulated by mitogens) from that of stimulated cells. The results showed that the HMB at concentrations 10, 50 and 100 µg HMB/ml of medium significantly (p < 0.05) increased T-lymphocyte proliferation stimulated by ConA and B-lymphocyte proliferation stimulated by LPS, respectively, at 20 °C and 30 °C, compared to control. The highest proliferative response of T-cells at 30 °C was observed at concentrations 50 and 100 µg HMB/ml in both species. The results showed that HMB stimulated the lymphocyte activity and suggested a possible application of HMB for modulation of cell-mediated immunity in sheatfish and catfish.

acta veterinaria brno

Effect of HMB (β-Hydroxy-β-Methylbutyrate) on in vitro Proliferative Responses of Sheatfish (Silurus glanis) and Catfish (Ictalurus melas) Lymphocytes Stimulated by Mitogens