Date of Award

12-2018

Culminating Project Type

Thesis

Degree Name

Biological Sciences - Cell and Molecular: M.S.

Department

Biology

College

College of Science and Engineering

First Advisor

Marina Cetkovic-Cvrlje

Second Advisor

Satomi Kohno

Third Advisor

Heiko L. Schoenfuss

Fourth Advisor

Nathan Bruender

Creative Commons License

Creative Commons License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License.

Keywords and Subject Headings

CEC, immune, toxicology, innate, fathead neutrophil

Abstract

Contaminants of emerging concern (CECs), including personal care products, pharmaceuticals, industrial agents and agricultural runoff, have shown adverse effects on reproduction and behavior in aquatic species, such as fathead minnow. Since the reproductive system and the immune system are tightly linked, it is critical to investigate effects of CECs on the immune system. An innate immunity, which is characterized by a specific cell type, neutrophils, represents an important branch of fish immune system. A decrease in innate immune functions could lead to an increase of infections, with a consequent impact on fish survival. We developed a neutrophil functional assay (myeloperoxidase degranulation assay, MPO), and a quantitative measurement of neutrophil-specific mRNA abundance (myeloid-specific peroxidase, elastase 2 and NADPH oxidase) by the reverse-transcription quantitative polymerase chain reaction (qPCR). Anterior kidneys, as a main source of fish neutrophils, were analyzed by the MPO assay and the qPCR after a 96-hour flow-through exposure of male adult fathead minnows to 21 individual compounds, which are commonly detected urban CECs, and their mixture. Chemical concentrations were based on those found in over 500 water samples collected by the United States Geological Survey as a part of the Great Lakes Restoration Initiative. A significant increase in degranulation was found in the preliminary experiments using estrogenic compounds, estrone and BPA, in particular concentrations. Urban CECs, such as ibuprofen, sulfamethoxazole and the urban mixture, increased the degranulation of neutrophils. Fexofenadine showed a significant increase in both degranulation and mpo mRNA abundance. 5-methyl-1H-benzotriazole was the only CEC to show a decrease in degranulation. Observed impact of studied CECs on the innate immune system might provide a novel insight in the ecotoxicology and expand our knowledge of CECs’ influence on the innate immune system of aquatic species.

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