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

Abstract

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.

Keywords

ammonia nitrate nitrification in vitro toxicity pharmacological agent fish therapeutics

Get full access to this article

View all access options for this article.

References

Verstraete

, & Alexander

(1973). Heterotrophic nitrification in samples of natural ecosystems. Environmental Science and Technology 1, 39–42.

Perfittini

, & Bianchi

(1990). The comparison of two simple protocols designed to initiate and stimulate ammonia oxidation in closed aqua-culture systems. Aquaculture 88, 179–188.

Sakai

, Ikehata

, Ikenaga

, Wakayama

, & Moriguchi

(1996). Nitrite oxidation by heterotrophic bacteria under various nutritional and aerobic conditions. Journal of Fermentation and Bioengineering 82, 613–617.

Noga

E.J.

(1996). Fish Disease: Diagnosis and Treatment, 377 pp. St Louis, MO, USA: Mosby Yearbook.

Carmignani

G.M.

, & Bennett

J.P.

(1977). Rapid start-up of biological filter in a closed aquaculture system. Aquaculture 11, 85–88.

Collins

M.T.

, Gratzek

J.B.

, Dawe

D.L.

, & Nemetz

T.G.

(1975). Effects of parasiticides on nitrification. Journal of the Fish Research Board of Canada 32, 2033–2037.

Collins

M.T.

, Gratzek

J.B.

, Dawe

D.L.

, & Nemetz

T.G.

(1976). Effects of antibacterial agents on nitrification in aquatic recirculating systems. Journal of the Fish Research Board of Canada 33, 215–218.

Bower

C.E.

, & Turner

D.T.

(1982). Effects of seven chemotherapeutic agents on nitrification in closed seawater culture systems. Aquaculture 29, 331–345.

Klaver

A.L.

, & Matthews

R.A.

(1994). Effects of oxytetracycline on nitrification in a model aquatic system. Aquaculture 123, 237–247.

10.

Gomez

, Mendez

, & Lema

J.M.

(1996). The effect of antibiotics on nitrification processes: batch assays. Applied Biochemistry and Biotechnology 57–58, 869–876.

11.

Benmoussa

, Martin

, Richard

, & Leprince

(1986). Inhibition of nitrification by heavy metal cations. Water Research 20, 1333–1339.

12.

Horrigan

S.G.

, Carlucci

A.F.

, & Williams

P.M.

(1981). Light inhibitions of nitrification in sea-surface films. Journal of Marine Research 39, 557–565.

13.

Olson

R.J.

(1981). Differential photoinhibition of marine nitrifying bacteria. Journal of Marine Research 39, 227–238.

14.

Moe

M.A.

Jr. (1992). The Marine Aquaculture Handbook: Beginner to Breeder, 512 pp. Plantation, FL, USA: Green Turtle Publications.

15.

US EPA (1985). Ambient Water Quality Criteria for Ammonia: 1984. EPA/440/5-95/001. Washington, DC, USA: Environmental Protection Agency.

16.

Ceriotti

(1974). Other non-protein nitrogenous compounds. In Clinical Biochemistry: Principles and Methods (ed. Curtius

H.C.

, & Roth

), pp. 1122–1126. Berlin, Germany: de Gruyter.

17.

Calleja

M.C.

, & Persoone

(1992). Cyst-based toxicity tests. IV. The potential of ecotoxicological tests for the prediction of acute toxicity in man as evaluated on the first ten chemicals of the MEIC programme. ATLA 20, 396–405.

18.

Isomaa

, Lilius

, & Rabergh

(1994). Aquatic toxicology in vitro. ATLA 22, 243–253.

19.

Kahru

, & Borchardt

(1994). Toxicity of 39 MEIC chemicals in bioluminescent photobacteria (the Biotox^™ test): correlation with other test systems. ATLA 22, 147–160.

20.

Scott

(1993). Therapy in aquaculture. In Aquaculture for Veterinarians: Fish Husbandry and Medicine (ed. Brown

), pp. 131–152. Oxford, UK: Pergamon Press.

21.

Sopinska

, Lutnicka

, & Guz

(1994). Studies on the influence of levamisole in carp after chronic intoxication with nitrogen compounds. Medycyna Weterynaryjna 12, 612–614.

22.

Griffin

B.R.

(1992). A simple procedure for identification of Cytophaga columnaris. Journal of Aquatic Animal Health 4, 63–66.

23.

Carson

, Schmidtke

L.M.

, & Munday

B.L.

(1993). Cytophaga johnsonae: a putative skin pathogen of juvenile farmed barramundi, Lates calcarifer Bloch. Journal of Fish Diseases 16, 209–218.

24.

Wang

P.C.

, & Liu

(1994). Characterization, survival conditions and drug susceptibility of Cytophaga columnaris isolated from diseased fish. Taiwanese Journal of Veterinary Medicine and Animal Husbandry 63, 85–95.

25.

Burka

J.F.

, Hammell

K.L.

, Horsberg

T.E.

, Johnson

G.R.

, Rainnie

D.J.

, & Speares

D.J.

(1997). Drugs in salmonid aquaculture: a review. Journal of Veterinary Pharmacology and Therapeutics 20, 333–349.

26.

Austin

(1985). Evaluation of antimicrobial compounds for the control of bacterial kidney disease in rainbow trout, Salmo gairdneri Richardson. Journal of Fish Diseases 8, 209–220.

27.

Hsu

H.M.

, Bowser

P.R.

, Schachte

J.H.

Jr. , Scarlett

J.M.

, & Babish

J.G.

(1995). Winter field trials of enrofloxacin for the control of Aeromonas salmonicoda infection in salmonids. Journal of the World Aquaculture Society 26, 307–314.

28.

Bowser

P.R.

, Wooster

H.A.

, & Hsu

H.M.

(1994). Laboratory efficacy of enrofloxacin for the control of Aeromonas salmonida infection in rainbow trout. Journal of Aquatic Animal Health 6, 288–291.

29.

Powell

M.D.

, Wright

G.M.

, & Speare

D.J.

(1995). Morphological changes in rainbow trout (Oncorhynchus mykiss) gill epithelia following repeated intermittent exposure to chloramine T. Canadian Journal of Zoology 1, 154–165.

30.

Flores Crespo

, Flores Crespo

, Ibarra Velarda

, Vera Montenegro

, & Vasquez Pelaez

(1995). Evaluation of chemotherapeutic drugs against cichlidogyriasis in tilapia fish (Oreochromis hornorum) in Mexico. Revista Latinoamericana de Microbiologia 37, 179–187.