Abstract: Identifying the effects of stressors on marine mammal health is critical for species conservation and collection. Adrenocorticotropic Hormone (ACTH) is a key indicator of peracute stress; however, handling methods may affect results. Thus, the aim of this study was to examine the effects of collection methods and storage temperature on the stability of bottlenose dolphin (Tursiops truncatus) ACTH measurements. Blood was collected from five managed-care bottlenose dolphins in four vacutainers™ in the following sequence: EDTA, sodium heparin, serum separator, and 2nd EDTA. The specimens were centrifuged, processed immediately, aliquoted into plasma and serum, and divided into four equal volumes. These aliquots were immediately stored at either 4º or -80º C for 24 h prior to the ACTH assay. ACTH concentrations were compared between the collection tube types and the storage temperatures. There was a significant difference in ACTH concentration across collection tube types. ACTH concentrations were highest in ethylenediamine-tetraacetic acid (EDTA) tubes and lowest in serum separator tubes at both temperatures. Twenty-four hour storage temperature did not have a statistically significant impact on mean ACTH concentration. There was no significant difference in ACTH concentration between the first and last drawn EDTA tubes. Bottlenose dolphin whole blood should be collected in EDTA for ACTH measurement. Samples may be refrigerated and shipped on cold packs if assayed within 24 h of collection or immediately frozen and shipped. When multiple samples are collected in various collection tubes at the same time, the order in which tubes are filled does not affect the ACTH measurement.
Key Words: Adrenocorticotropic Hormone, ACTH, collection method, anticoagulant, EDTA, heparin, temperature, bottlenose dolphins, Tursiops truncates
Document Type: Research article
DOI: 10.1578/AM.39.4.2013.324
Page Numbers: 324-329