title: Pathology working group review of histopathologic specimens from three laboratory studies of diclofenac in trout.
abstract: While the pathology peer review/pathology working group (PWG) model has long been used in mammalian toxicologic pathology to ensure the accuracy, consistency, and objectivity of histopathology data, application of this paradigm to ecotoxicological studies has thus far been limited. In the current project, the PWG approach was used to evaluate histopathologic sections of gills, liver, kidney, and/or intestines from three previously published studies of diclofenac in trout, among which there was substantial variation in the reported histopathologic findings. The main objectives of this review process were to investigate and potentially reconcile these interstudy differences, and based on the results, to establish an appropriate no observed effect concentration (NOEC). Following a complete examination of all histologic sections and original diagnoses by a single experienced fish pathologist (pathology peer review), a two-day PWG session was conducted to allow members of a four-person expert panel to determine the extent of treatment-related findings in each of the three trout studies. The PWG was performed according to the United States Environmental Protection Agency (US EPA) Pesticide Regulation (PR) 94-5 (EPA Pesticide Regulation, 1994). In accordance with standard procedures, the PWG review was conducted by the non-voting chairperson in a manner intended to minimize bias, and thus during the evaluation, the four voting panelists were unaware of the treatment group status of individual fish and the original diagnoses associated with the histologic sections. Based on the results of this review, findings related to diclofenac exposure included minimal to slightly increased thickening of the gill filament tips in fish exposed to the highest concentration tested (1,000 μg/L), plus a previously undiagnosed finding, decreased hepatic glycogen, which also occurred at the 1,000 μg/L dose level. The panel found little evidence to support other reported effects of diclofenac in trout, and thus the overall NOEC was determined to be >320 μg/L. By consensus, the PWG panel was able to identify diagnostic inconsistencies among and within the three prior studies; therefore this exercise demonstrated the value of the pathology peer review/PWG approach for assessing the reliability of histopathology results that may be used by regulatory agencies for risk assessment.
authors.affiliation: Experimental Pathology Laboratories, Inc., Sterling, VA, USA. Electronic address: jwolf@epl-inc.com.
authors.forename: Jeffrey C
authors.initials: JC
authors.lastname: Wolf
authors.affiliation: Bayer HealthCare AG, Wuppertal, Germany.
authors.forename: Christine
authors.initials: C
authors.lastname: Ruehl-Fehlert
authors.affiliation: Centre for Fish and Wildlife Health, University of Bern, Bern, Switzerland.
authors.forename: Helmut E
authors.initials: HE
authors.lastname: Segner
authors.affiliation: AnaPath GmbH, Oberbuchsiten, Switzerland.
authors.forename: Klaus
authors.initials: K
authors.lastname: Weber
authors.affiliation: Experimental Pathology Laboratories, Inc., Research Triangle Park, NC, USA.
authors.forename: Jerry F
authors.initials: JF
authors.lastname: Hardisty
mesh_terms: D000818:Animals; D004008:Diclofenac / Q000633:toxicity*; D005880:Gills / Q000187:drug effects*; D007668:Kidney / Q000187:drug effects*; D008099:Liver / Q000187:drug effects*; D017686:Oncorhynchus mykiss / Q000502:physiology*; D010380:Peer Review; D015203:Reproducibility of Results; D014874:Water Pollutants, Chemical / Q000633:toxicity*
chemical_list: D014874:Water Pollutants, Chemical; D004008:Diclofenac
keywords: DCF; Diclofenac; Fish; Histopathology; LOEC; NOEC; PWG; Pathology Working Group; Peer review; RP; SP; Trout; diclofenac; lowest observed effect concentration; no observed effect concentration; pathology working group; reviewing pathologist; study pathologist