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ToxSci Advance Access published online on August 13, 2004
Toxicological Sciences, doi:10.1093/toxsci/kfh250
Toxicological Sciences © Society of Toxicology 2004; all rights reserved
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Received June 25, 2004
Accepted July 30, 2004

Environmental Toxicology

Influence of Chronic Exposure to Treated Sewage Effluent on the Distribution of White Blood Cell Populations in Rainbow Trout (Oncorhynchus mykiss) Spleen

Birgit Hoeger 1, Bernd Koellner 2, Guenter Kotterba 2, Michael R. van den Heuvel 3, Bettina Hitzfeld 4, Daniel R. Dietrich 1*

1 Environmental Toxicology, University of Konstanz, P.O.Box X918, D-78457 Konstanz, Germany
2 Federal Research Centre for Virus Diseases in Animals, Greifswald-Insel Riems, Germany
3 Forest Research, Private Bag 3020, Sala St., Rotorua, New Zealand
4 Swiss Agency for the Environment, Forests and Landscape SAEFL, Substances, Soil, Biotechnology Division, Section Substances, 3003 Bern, Switzerland

* To whom correspondence should be addressed. E-mail: daniel.dietrich{at}uni-konstanz.de.


  Abstract

Impairment of immune function in aquatic animals has been proposed as a possible consequence of low-level contamination of surface waters with anthropogenic substances such as through the discharge of wastewater into rivers, lakes and oceans. The study at hand investigated the effects of chronic (32 weeks) exposure to sewage treatment plant (STP) effluent on the prevalence and distribution of different leucocyte populations in spleen samples of rainbow trout (Oncorhynchus mykiss). To simulate an infection, fish were injected intraperitoneally (ip) with inactivated Aeromonas salmonicida salmonicida, six weeks prior to the termination of the experiment. Immunohistological analysis revealed a marked decrease in thrombocyte numbers, an increase of monocytes, altered distribution of B-cells and higher surface immunoglobulin expression, as well as activation of MHC class II expression in the spleen after exposure to 15% (v/v) effluent. The most prominent finding of the present study, however, was the occurrence of intraplasmatic deposits or inclusions with strong autofluorescence in spleen sections from effluent-exposed trout. In addition to effluent effects, injection of trout with A. salmonicida stimulated infiltration of monocytes, increased staining intensity on thrombocytes and enhanced MHC class I expression in larger leucocytes surrounding melanomacrophage centres. In general, the current study demonstrates a marked, potentially adverse effect of STP effluent on spleen leucocytes and on the integrity of spleen tissue. The observed response suggests a constant unspecific stimulation of different leucocyte populations and is reminiscent of chronic inflammation.

Keywords: Oncorhynchus mykiss; fish immune system; fish immunology; Aeromonas salmonicida; trout leucocytes; trout specific monoclonal antibodies.
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