DAO 95:73-79 (2011)  -  doi:10.3354/dao02333

Development and maintenance of a specific pathogen-free (SPF) zebrafish research facility for Pseudoloma neurophilia

Michael L. Kent1,*, Cari Buchner2, Virginia G. Watral1, Justin L Sanders1, Jane LaDu2, Tracy S. Peterson1, Robert L. Tanguay2

1Department of Microbiology, and 2Department of Environmental and Molecular Toxicology, Oregon State University, Corvallis, Oregon, USA

ABSTRACT: Pseudoloma neurophilia (Microsporidia) is very common in zebrafish Danio rerio research facilities. A new zebrafish facility has been established at the Sinnhuber Aquatic Resource Laboratory (SARL), Oregon State University, Corvallis, OR, USA, and this was an opportunity to establish a specific pathogen-free (SPF) colony of zebrafish for this microsporidium. Progeny from 9 zebrafish lines (n = 2203) were initially transferred to the SARL facility in 2007 following PCR screening of broodstock and a subpopulation of progeny (258 of 1000 fish from each family). Screening of fish for P. neurophilia within the facility was conducted as follows: (1) Moribund or dead fish were examined by histology. (2) Each line was regenerated on a 4 mo rotation, and a subsample of each of these major propagations (60 fry, in pools of 10) was PCR-screened at 10 d post hatch. (3) Adult fish (~1 yr old) from each line were euthanized; 20 fish were examined by histology and the brains of another 60 fish (in pools of 5) were screened by PCR. (4) This screening was replicated on sentinel fish held in 4 tanks receiving effluent water from all tanks in the facility (20 fish per tank). (5) Four-month old fish (n = 760) from a toxicology study conducted within the laboratory were examined by histology. To date, we have evaluated 2800 fish by PCR and 1222 fish by histology without detecting P. neurophilia. Thus, we have established 9 lines of zebrafish SPF for P. neurophilia. However, 26 fish exhibited mycobacteriosis, with acid-fast bacteria present in tissue sections, and 49 other fish had incidental lesions.

KEY WORDS: Zebrafish · Disease control · Microsporidia

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Cite this article as: Kent ML, Buchner C, Watral VG, Sanders JL, LaDu J, Peterson TS, Tanguay RL (2011) Development and maintenance of a specific pathogen-free (SPF) zebrafish research facility for Pseudoloma neurophilia. Dis Aquat Org 95:73-79

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