In the present study, we investigated the in vitro immunostimulatory effects of inactivated form of two potential probiotics, Bacillus amyloliquefaciens FPTB16 and B. subtilis FPTB13 in catla (Catla catla). Catla head kidney leukocytes (HK) were incubated with the bacteria alive or inactivated with heat shock (2 h at 60 °C), UV light (for 2.5 h) and formalin (1.0%, v/v) treatment (for 24 h at 4 °C) at different concentrations (10(7), 10(8) and 10(9) cells ml(-1)). After incubation, different cellular immune parameters such as respiratory burst activity, nitric oxide production, leukocyte peroxidase content and proliferative response were analyzed. The inactivated probiotic preparations stimulated all the cellular immune parameters of catla HK leukocytes in vitro. Among the different inactivated preparations, heat-treated form exhibited the best result. The lowest dose (10(7) cells ml(-1)) of both the strains showed the maximum stimulation. The results collectively suggest the efficacy of inactivated preparations to be used as immunostimulant in aquaculture.

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Immunostimulants increase the immunocompetency and disease resistance of fish by enhancing immune responses. This study determined the in vitro and dietary effects of chitin on immune responses of catla, Catla catla. Catla head-kidney leukocytes were cultured in complete RPMI-1640 medium containing 0.01, 0.1, and 1.0 mg/mL of chitin. Leucocyte viability and different cellular immune responses, namely, respiratory burst, nitric oxide production, and leucocyte peroxidase content were analyzed in vitro. In the dietary experiment, chitin-supplemented diets (0.5, 1.0, and 2.0% of feed) were fed to catla for 2 wk, and respiratory burst, serum lysozyme, and total protein contents were measured after 1 and 2 wk of feeding. Leucocytes incubated in vitro with chitin for 72 h maintained their viability. Chitin significantly (P < 0.05) enhanced all the immune responses in vitro. The lowest dose showed the best response. Dietary chitin also significantly stimulated all the immune parameters in a dose-dependent manner with the highest dose showing the maximum responses. All the responses were higher after 1 wk of feeding and declined thereafter. The results suggest that chitin stimulates catla cellular and humoral immune responses in vitro and in vivo.