Exposure to Paper Mill Effluent at a Site in North Central Florida Elicits Molecular-Level Changes in Gene Expression Indicative of Progesterone and Androgen Exposure

Abstract

Endocrine disrupting compounds (EDCs) are chemicals that negatively impact endocrine system function, with effluent from paper mills one example of this class of chemicals. In Florida, female Eastern mosquitofish (Gambusia holbrooki) have been observed with male secondary sexual characteristics at three paper mill-impacted sites, indicative of EDC exposure, and are still found at one site on the Fenholloway River. The potential impacts that paper mill effluent exposure has on the G. holbrooki endocrine system and the stream ecosystem are unknown. The objective of this study was to use gene expression analysis to determine if exposure to an androgen receptor agonist was occurring and to couple this analysis with in vitro assays to evaluate the presence of androgen and progesterone receptor active chemicals in the Fenholloway River. Focused gene expression analyses of masculinized G. holbrooki from downstream of the Fenholloway River paper mill were indicative of androgen exposure, while genes related to reproduction indicated potential progesterone exposure. Hepatic microarray analysis revealed an increase in the expression of metabolic genes in Fenholloway River fish, with similarities in genes and biological processes compared to G. holbrooki exposed to androgens. Water samples collected downstream of the paper mill and at a reference site indicated that progesterone and androgen receptor active chemicals were present at both sites, which corroborates previous chemical analyses. Results indicate that G. holbrooki downstream of the Fenholloway River paper mill are impacted by a mixture of both androgens and progesterones. This research provides data on the mechanisms of how paper mill effluents in Florida are acting as endocrine disruptors.

Degradation of Bile Acids by Soil and Water Bacteria

“…Although only little information is available regarding the endocrine effects of hydroxylated ADDs and ADs, they might also have androgenic potential, since exogenous steroids with similar structures to testosterone affect the androgen signaling pathways in vertebrates [ 164 , 165 ]. Similarly, endocrine effects of bacterial sterol degradation on fish were suggested: masculinization of fish populations in rivers receiving paper-mill effluents [ 166 , 167 , 168 ] was traced back to the bacterial transformation of plant-derived sterols into androgenic steroids [ 169 , 170 , 171 ].…”

Progesterone: A Steroid with Wide Range of Effects in Physiology as Well as Human Medicine

“…They have been detected in wastewater and surface waters, livestock manure, soil, and the runoff from farms, and can accumulate in water sediments. They have also been found in paper mill effluents, as steroids can be released during the processing of pine pulp or through the microbial degradation of phytosteroids present in the waste from processing pine trees [ 257 , 258 , 259 ]. Due to their hydrophobic nature, progestins are able to accumulate in aquatic organisms [ 260 ].…”

Estudio de la perturbación del desarrollo sexual de Cnesterodon decemmaculatus (Pisces, Poeciliidae) por compuestos con actividad estrogénica

“…Aunque el término "perturbación endócrina" ha sido introducido hace casi tres décadas, existen numerosos antecedentes de perturbación endócrina en organismos acuáticos, incluyendo invertebrados y vertebrados (Sumpter, 2002). Los efectos documentados en organismos acuáticos expuestos a EDCs, incluyen desde imposex inducido por tributilo de estaño (TBT) en moluscos marinos hasta la masculinización o feminización de algunas especies de peces que habitan aguas abajo de la zona de descarga de efluentes de la industria papelera (Bryan et al, 1986;Cody y Bortone, 1997;Brockmeier et al, 2014) y de efluentes cloacales (Purdom et al, 1994;Jobling et al, 2006), respectivamente.…”

“…Por ejemplo, en el caso de poecílidos machos de los géneros Gambusia y Xiphophorus, se ha mostrado que el desarrollo del gonopodio puede ser alterado por concentraciones ambientalmente relevantes de EDCs en el agua, como el 4-nonilfenol (Drèze et al, 2000), E2 (Doyle y Lim, 2002) y EE2 (Angus et al, 2005). También, se ha demostrado la capacidad de los EDCs de inducir masculinización de la aleta anal de las hembras expuestas a efluentes de la industria papelera (Drysdale y Bortone, 1989;Brockmeier et al, 2014), 11-KT (Angus et al, 2001), androstenediona (Stanko y Angus, 2007), levonorgestrel (Frankel et al, 2016b), 17β-trembolona (Brockmeier et al, 2013), etiniltestosterona y 17αmetiltestosterona (Turner, 1942b;Amiri-Moghaddam et al, 2010). De esta forma, la estructura del gonopodio ha demostrado ser un indicador útil de efecto adverso inducido por los EDCs y su análisis no requiere de infraestructura sofisticada ni de reactivos costosos.…”

“…han reportado efectos antiandrogénicos en ejemplares silvestres de Gambusia holbrooki que habitaban en un lago de Estados Unidos. Así como también, se han documentado efectos androgénicos en poblaciones silvestres de Gambusia holbrooki que habitaban en cuerpos de agua dulce impactados por efluentes de la industria papelera en Estados Unidos (Howell et al, 1980;Bortone et al, 1989;Cody y Bortone, 1997;Bortone y Cody, 1999;Orlando et al, 2002;Brockmeier et al, 2014).…”