Interactive toxic effects of heavy metals and humic acids on Vibrio
fischeri
V. Tsiridis1,2, M. Petala1,2, P. Samaras3, S. Hadjispyrou4, G. P. Sakellaropoulos1,2 and A.
Kungolos5*
1
Department of Chemical Engineering, Aristotle University of Thessaloniki, 54006, Thessaloniki, Greece
2
Chemical Process Engineering Research Institute, 6th km Harilaou Thermi Road, 57001, Thessaloniki,
Greece
3
Department of Pollution Control Technologies, Τechnological Education Institution of West Macedonia,
50100, Kozani, Greece
4
Department of Chemical Engineering, Aristotle University of Thessaloniki, 54006, Thessaloniki, Greece
5
Department of Planning and Regional Development, University of Thessaly, 38334 Volos, Greece
*Corresponding author: E-mail: kungolos@uth.gr, Tel +30 24210 74480, Fax: +302421074380
Abstract
The presence of heavy metals in solid and liquid wastes is an important issue related to the pollution
of the environment. It is generally accepted that the solubility, the bioavailability and the toxicity of
heavy metals are dependent on various physicochemical parameters such as pH, hardness,
interactive effects and the presence of natural organic matter. Additionally, heavy metals are often
present in the environment in mixtures, making the task of the assessment of the environmental
hazard even more difficult, due to the antagonistic or synergistic actions that may occur. The
investigation of the joint toxic effects of the chemicals in a mixture is generally based on the
comparison of the actual toxic effect of the mixture with the theoretically expected toxic effect
deduced by a statistical model, using the toxic effects of the individual chemicals. The
bioavailability of metals may be affected by the presence of natural organic matter, such as humic
acids, which are produced by the degradation of dead organic materials. The effect of humic acids
(HAs) on the toxicity of copper and zinc, and the effects of HAs on metal toxicities were
investigated using the photobacterium Vibrio fischeri as a test organism. The solutions with an
initial concentration 10 mg/L of the tested metals were freshly prepared by dissolving the following
chloride salts in deionized water: zinc chloride (ZnCl2) and copper chloride dehydrated
(CuCl2.2H2O). The effect of humic acids (HA) on the toxicity of the tested heavy metals was
evaluated using 1, 10 and 20 mg/L humic acids that were prepared by a stock humic acids (Fluka,
Germany) solution of 100 mg/L. All chemicals used were of analytical grade. Furthermore, the pH
value of all metal solutions was adjusted to 7  0.2, prior to the toxicity tests, by the addition of
0.1N HCl or 0.1N NaOH solutions.
The complexation of heavy metals with humic acids is an important parameter that may alter the
toxicity and bioavailability of heavy metals in the aquatic system. The examination of the
complexation of heavy metals with humic acids showed that the toxicity of Cu decreased with the
addition of humic acids, while the toxicity of Zn remained almost constant. The interactive effects
of Cu and Zn mixtures on V. fisheri were found to be synergistic. The presence of HA in the binary
mixtures caused relatively high toxicity reduction. It was found that the presence of humic acids
reduced the toxicity of Cu and Zn applied together. It can also be concluded that, for the proper
estimation of the environmental hazard caused by a chemical, the presence of other constituents that
may interact with the chemical of concern should be evaluated.
Keywords: Microtox; heavy metals; interactive effect; bioavailability; toxicity.