Isolation of Compounds From Bleached Kraft Mill Recovery Condensates Associated With Reduced Levels of Testosterone in Mummichog (Fundulus Heteroclitus)

Abstract

Previous studies have identified chemical recovery condensates as a primary source of hormonally active substances within a Canadian bleached kraft pulp mill. Although reverse osmosis treatment of condensates raises the exposure threshold that alters circulating levels of testosterone in mummichog (Fundulus heteroclitus), the responsible chemicals have not been characterized. In this study, a solid-phase extraction (SPE) method was developed to isolate chemical recovery condensate extractives for evaluation of hormonal activity. Condensates generated during softwood and hardwood pulp production were investigated for their relative potential to affect both circulating and gonadal production of sex steroids in mummichog. Mummichog were exposed to whole condensates, extracts from suspended particulates (> 1 microm), two fractions from SPE, and residual condensates after SPE. The distribution of bioactivity among condensate fractions was similar for both wood furnishes. In both sexes, significant depressions in circulating testosterone and in in vitro gonadal testosterone production were associated with exposure to whole condensates, particulate extracts, one SPE fraction, and residual material after SPE. An optimized SPE method subsequently demonstrated complete recovery of polar, bioavailable chemicals that reduced testosterone levels in both sexes. Characterizations of active fractions by gas chromatography-mass spectrometry (GC-MS) showed the presence of extractives with molecular masses > or = 240 amu possessing functionalities consistent with lignin degradation products. This study provides the first isolation of chemicals derived from pulp production associated with impaired reproductive performance in fish.

Detectable effect size and bioassay power of mummichog (Fundulus heteroclitus) and fathead minnow (Pimephales promelas) adult reproductive tests

“…Testosterone was unaffected in male and female mummichog Table 2. Testosterone depressions were observed in male and female mummichog exposed for 30 or 60 d to 50% CME following installation of a reverse-osmosis effluent treatment system [27,33] and in females at 1 and 50% concentrations [26]. b Females (F) exposed to 3% effluent weighed significantly more than did controls ( p 5 0.02).…”

Pulp and paper mill effluent treatments have differential endocrine‐disrupting effects on rainbow trout

“…Toxicol. Canadian kraft bleached mill condensates (condensed vapor produced when black liquor is concentrated in evaporators as part of the chemical recovery process) were solid-phase extracted (similar to the methods used in our experiment) and used to evaluate the association with testosterone reduction in a waterborne exposure experiment using adult male and prespawning female mummichog (Fundulus heteroclitus) [36]. 28, 2009 187 that provide mechanistic data, the high variability and discrepancies in relative potency of the in vitro tests seem to be associated with the lower level of biological organization used in each assay, thus having an intrinsic limitation in trying to elucidate environmentally relevant effects [8,33].…”

Development of a short‐term reproductive endocrine bioassay using steroid hormone and vitellogenin end points in the estuarine mummichog (Fundulus heteroclitus)

“…As well, basal steroid levels were often at the detection limits of the RIA. Medium 199 has been shown to be an effective medium for gonadal incubations in this species [10; this study] and the incubation requirements have been further optimized here from that earlier work. Basal testes testosterone production generally yields testosterone levels at or below 1 pg/mg tissue/h [9,10; this study, Fig.…”