The effect of peroral administration of toxic cyanobacteria on laboratory rats (Rattus norvegicus var. alba).

OBJECTIVES: The toxic cyanobacteria are a serious problem for water supply systems, recreation, and agriculture. Cyanobacteria produce numerous bioactive compounds including microcystins - the most studied cyanobacterial hepatotoxins. Only rare studies addressed realistic situation, i.e. impact of MCs accumulated in the fish tissues on the overall physiology. The aim of the present study was to provide a model simulation of the simple food chain for evaluation of impacts of cyanobacteria on the rat physiology under different exposure scenario. METHODS: Experimental rats were fed with food with fish meat, which contained external additions of isolated microcystins as well as toxic cyanobacteria Microcystis, nontoxic cyanobacteria Arthrospira and green alga Chlorella. Subgroups of the animals were also challenged with a model antigen KLH to investigated immune-related parameters. We studied parameters of oxidative stress in the liver as levels of lipid peroxidation and glutathion levels. Series of hematological, biochemical and immunological parameters were also investigated. RESULTS: Although considerable amounts of microcystins were administered to rats, all levels of MCs were under the detection limit (1 ng/g fresh weight) in the rat tissues using tandem LC/MS. Only some conjugates of microcystins with cystein and glutathion were detected in the rat liver exposed to Microcystis biomass (values were around the detection limit). Statistically significant depletion of body and liver weight was observed in groups with microcystin addition in comparison with all other groups. Rats exposed to MCs had stimulated immune system (showed higher antibody answer on administered antigen). Also modulation of some lymphocyte subpopulations was recorded with the most interesting observation of stimulated NK cell numbers in groups exposed to isolated toxins (but not to biomass containing the same toxin amount). CONCLUSIONS: Our study demonstrates that oral exposure to microcystins in the diet may induce some detoxification responses and modulation of some hematological and immunological parameters.

Effect of mitogen lectin on lymphocyte or brain cortex cell activation.

OBJECTIVES: We studied a) mitogen lectin (PHA) evoked changes of Na+/K+-ATPase activity in functionally different lymphocytes or brain cortex cells and b) quantitative relationship between PHA- evoked early enzyme activation and late lymphocyte proliferation were analyzed. MATERIALS AND METHODS: We performed biochemical analyses of Pi released from ATP by Na+/K+-ATPase activity. Lymphocyte proliferation was assayed by 3H-thymidine incorporation. RESULTS: We demonstrated PHA stimulated Na+/K+-ATPase activity of mouse spleen lymphocytes or freshly isolated brain cortex cells. Besides this, we estimated high stimulation of Na+/K+-ATPase activity and subsequent late 3H-thymidine incorporation into pig lymphocytes as both PHA dose and K+ ion concentration dependent. CONCLUSIONS: Thus, early PHA dose-dependent stimulation of Na+/K+-ATPase activity is a more general response in different animal species and functionally different cells. We measured both cell type- and PHA-dose dependent enzyme activity stimulation. We can suggest that intensity of early PHA induced Na+/K+-ATPase activation could be in relationship to subsequent elevated level of T lymphocyte proliferation. The Na+/K+-ATPase can be a part of mitogen lectin evoked signal transduction mechanisms.