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Alternatives to Laboratory Animals - ATLA

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Short-term Toxicity of Various Pharmacological Agents on the In Vitro Nitrification Process in a Simple Closed Aquatic System

Herman Nimenya, Annie Delaunois, Duc La Duong, Serge Bloden, Jean Defour, Baudouin Nicks and Michel Ansay

During the treatment of fish diseases, drugs which inhibit the nitrification process can cause acute ammonia toxicity. The same phenomenon can occur when fish are put into a tank without active cultures of nitrifying bacteria. The purpose of this study was to quantify the inhibitory effects of 15 pharmacological agents, which are often used as therapeutic agents in ichthyopathology, on ammonia removal and nitrate production in a simple closed aquatic system. The experiments were conducted in polyethylene bags containing activated biofilters and synthetic water solutions, held in a water bath. Ammonia was added to initiate the nitrification process, and graded concentrations of various pharmacological agents were added. The effects of the pharmacological agents on in vitro nitrification were assessed by monitoring ammonia and nitrate concentrations compared to controls with no added agents, for 24 hours. Graded concentrations of ampicillin (Albipen®), chloramine T, enrofloxacin (Baytril®), erythromycin, levamisole, methylene blue and polymyxin B induced dose-dependent inhibitions of ammonia removal and nitrate production. The corresponding linear regression curves showed high correlation coefficients and were highly significant (p < 0.05). The addition of chloramphenicol, copper (II) sulphate, kanamycin disulphate, malachite green, neomycin sulphate, potassium penicillin G, tetracycline and a mixture of trimethoprim and sulphadoxin (Duoprim™) had no significant effects on the nitrification process. A significant dose-related inhibition of nitrate production, but not of ammonia oxidation, was observed with enrofloxacin. The significant correlation (r = 0.940; p < 0.001) between the degrees of inhibition of ammonia oxidation and nitrate production for the various inhibitory pharmacological agents has also been calculated, with a view to validating this method. The data presented suggest that separate tank facilities for hospitalisation or quarantine are necessary when treating diseased fish with ampicillin, enrofloxacin, chloramine T, erythromycin, levamisole, methylene blue or polymyxin B, in order to avoid ammonia poisoning.