Gene transcription in sea otters (Enhydra lutris); development of a diagnostic tool for sea otter and ecosystem health

LIZABETH BOWEN, A. KEITH MILES, MICHAEL MURRAY, MARTIN HAULENA, JUDY TUTTLE, WILLIAM VAN BONN, LANCE ADAMS, JAMES L. BODKIN, BRENDA BALLACHEY, JAMES ESTES, M. TIM TINKER, ROBIN KEISTER, JEFFREY L. STOTT

Gene transcription analysis for diagnosing or monitoring wildlife health requires the ability to distinguish pathophysiological change from natural variation. Herein, we describe methodology for the development of quantitative real-time polymerase chain reaction (qPCR) assays to measure differential transcript levels of multiple immune function genes in the sea otter (Enhydra lutris); sea otter-specific qPCR primer sequences for the genes of interest are defined. We establish a 'reference' range of transcripts for each gene in a group of clinically healthy captive and free-ranging sea otters. The 10 genes of interest represent multiple physiological systems that play a role in immuno-modulation, inflammation, cell protection, tumour suppression, cellular stress response, xenobiotic metabolizing enzymes, antioxidant enzymes and cell-cell adhesion. The cycle threshold (C(T)) measures for most genes were normally distributed; the complement cytolysis inhibitor was the exception. The relative enumeration of multiple gene transcripts in simple peripheral blood samples expands the diagnostic capability currently available to assess the health of sea otters in situ and provides a better understanding of the state of their environment.