Sample storage conditions can affect accuracy and reproducibility of biological measurements. Storing samples rapidly at the lowest available temperatures is considered ideal but is not always feasible when sampling in remote and logistically challenging field conditions, as is often the case with sea turtles. The objective of this study was to examine the stability of plasma proteins and quality of whole blood RNA from loggerhead sea turtle samples collected as part of an eighteen-year-long curated specimen collection. These biological variables are often used to assess sea turtle health; therefore, it is necessary to maintain the integrity of these components during storage. Protein electrophoresis was conducted on heparinized plasma from individual turtles collected in 2018 (n = 3), 2008 (n = 3), and 2001 (n = 3). Plasma was also pooled from four turtles sampled in 2018 and subjected to various storage temperatures. Whole blood was collected in blood collection tubes containing sodium heparin or PAXgene tubes with an RNA preservative. These were subjected to different storage treatments that can possibly occur during logistically difficult field sampling. Following various treatments, plasma proteins showed minor differences across collection years and no differences among storage treatments were observed, even when exposed to 38°C for three hours. RNA quality was assessed from whole blood using an RNA integrity number (RIN). RINs were poor from sodium heparin tubes that were frozen and from PAXgene tubes after an extended thaw. High-quality RNA was obtained from sodium heparin tubes that were never frozen and from PAXgene tubes with freezing delayed by up to 11 days. Overall, these results indicate that plasma proteins remain stable over time and when exposed to undesirable storage conditions, and RNA degrades rapidly in sea turtle blood after freezing and when not properly preserved. These aspects are important to consider when planning sampling protocols and logistics for optimal long-term sample preservation.

中文翻译：

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测试红海龟存档血液中血浆蛋白和全血 RNA 的稳定性，Caretta caretta。

样品储存条件会影响生物测量的准确性和重现性。在最低可用温度下快速储存样品被认为是理想的，但在偏远且物流困难的野外条件下取样时并不总是可行的，海龟通常就是这种情况。本研究的目的是检查来自红海龟样本的血浆蛋白的稳定性和全血 RNA 的质量，这些样本是为期 18 年的精选标本收集的一部分。这些生物变量通常用于评估海龟的健康状况；因此，有必要在存储过程中保持这些组件的完整性。对2018年（n  = 3）、2008年（n = 3) 和 2001 ( n = 3)。血浆还来自 2018 年采样的四只海龟，并经受了不同的储存温度。全血收集在含有肝素钠或 PAXgene 管和 RNA 防腐剂的采血管中。这些经历了不同的储存处理，这些处理可能在后勤困难的现场采样期间发生。在经过各种处理后，血浆蛋白在不同收集年份之间显示出微小差异，并且即使在 38°C 下暴露 3 小时，也没有观察到储存处理之间的差异。使用 RNA 完整性数 (RIN) 从全血中评估 RNA 质量。冷冻的肝素钠管和延长解冻后的 PAXgene 管的 RIN 较差。从从未冷冻的肝素钠管和冷冻延迟长达 11 天的 PAXgene 管中获得高质量的 RNA。总体而言，这些结果表明血浆蛋白随着时间的推移和暴露于不良储存条件下保持稳定，而海龟血液中的 RNA 在冷冻后和未正确保存时会迅速降解。在规划采样方案和物流以实现最佳的长期样品保存时，这些方面是重要的考虑因素。