Comparison of Selected Nonlethal Samples from Adult Steelhead for Detection of Infectious Hematopoietic Necrosis Virus Using Cell Culture

Abstract

Nonlethal sampling techniques have previously been evaluated for detection of a variety of viral salmonid pathogens. However, many of these studies have used molecular assays in lieu of widely accepted cell culture techniques to evaluate the sampled tissues. Samples were collected from female steelhead Oncorhynchus mykiss broodstock using three potential nonlethal sampling methods (mucus/skin scrape, pectoral fin clip, and gill tissue biopsy) and evaluated for the presence of infectious hematopoietic necrosis virus (IHNV) via cell culture techniques. The results were compared with those from samples collected using a standard lethal sampling method (pooled anterior kidney and spleen tissues) applied to the same fish. Of the three nonlethal sampling techniques that were evaluated, fin clipping was the easiest and least invasive method. Furthermore, fin tissue was as sensitive as or more sensitive than kidney/spleen tissue for detecting IHNV in this population of fish. However, with the exception of gill tissue, the nonlethal samples did not appear to be appropriate surrogates for lethally collected tissues with regard to identifying an active infection in a particular fish. Nevertheless, nonlethal sampling coupled with cell culture appears to be suitable for helping to define the IHNV status of a steelhead population. Received July 27, 2016; accepted December 11, 2016.

A Comparison of Nonlethal and Destructive Methods for Broad‐Based Infectious Agent Screening of Chinook Salmon Using High‐Throughput qPCR

“…The importance of logistical constraints regarding nonlethal tissue sampling (e.g., the invasiveness of procedures) has been characterized for single agent assessments, with the benefits of some less invasive techniques (e.g., fin clipping) providing insufficient information on host health and infection development (Burbank et al. ). In this study, the processing of blood was more time‐consuming than of gills, taking into account the dilution process to avoid downstream issues with protein contamination in purified RNA as well as troubleshooting the clotting issues following the thawing of samples.…”

Infectious hematopoietic necrosis virus: advances in diagnosis and vaccine development

“…Recently, Burbank, Fehringer & Chiaramonte (2017) reported a non-lethal sampling technique through fin clipping in adult steelhead trout, followed by the detection of IHNV with cell culture techniques. This method has been demonstrated to be more efficient than the standard lethal sampling methods, such as spleen and anterior kidney sampling (Burbank, Fehringer & Chiaramonte, 2017).…”

“…Recently, Burbank, Fehringer & Chiaramonte (2017) reported a non-lethal sampling technique through fin clipping in adult steelhead trout, followed by the detection of IHNV with cell culture techniques. This method has been demonstrated to be more efficient than the standard lethal sampling methods, such as spleen and anterior kidney sampling (Burbank, Fehringer & Chiaramonte, 2017). The confirmation test or “gold standard” for IHNV diagnosis is by detecting the virus in cell cultures, followed by diagnosis using immunological and molecular techniques (Barlič-Maganja et al, 2002; Burbank, Fehringer & Chiaramonte, 2017; Crane & Williams, 2008; Winton, 1991; Woo, Leatherland & Bruno, 2011).…”

Peer Review #2 of "Infectious hematopoietic necrosis virus: advances in diagnosis and vaccine development (v0.1)"

“…Recently, Burbank et al (2017) reported a non-lethal sampling technique through fin clipping in adult steelhead trout, followed by the detection of IHNV with cell culture techniques.…”

“…This method has been demonstrated to be more efficient that the standard lethal sampling methods, such as spleen and anterior kidney sampling (Burbank et al 2017). The confirmation test or 'gold standard' for IHNV diagnosis is by detecting the virus in cell cultures, followed by diagnosis using immunological and molecular techniques (Barlič-Maganja et al 2002;Burbank et al 2017;Crane 2008;Winton 1991;Woo et al 2011). The presence of IHNV is routinely assessed by observing the development of viral cytopathic effect (CPE) in cell lines such as epithelioma papulosum cyprinid (EPC) and fathead minnow (FHM) under a phase-contrast microscope (Dixon et al 2016).…”