Abstract The eastern slope of the Grand Banks, Newfoundland and Labrador, Canada, represents an area where active seismic exploration overlaps extensively with an important snow crab (Chionoecetes opilio) fishery. Field studies were conducted in 2016 and 2017 to investigate the potential impact of seismic oil and gas surveying exposure on the transcriptome of the snow crab hepatopancreas, which is involved in metabolism and response to environmental stress. In 2016, snow crabs were subjected to 2D seismic noise for 2 h and sampled before (i.e. pre-seismic), and 18 h and 3 weeks after exposure in Carson Canyon (CC). In 2017, the 2D seismic exposure was repeated and samples were collected before and 1 day, 2 days, and 6 weeks after exposure in CC. Additionally, in 2017, snow crabs were subjected to 3D seismic noise for 2 months and sampled during and 6 weeks after exposure at a site north of CC. In both years, snow crabs were also collected from a separate non-seismic exposed site [Lilly Canyon (LC)]. RNA-seq was used to identify candidate seismic-responsive molecular biomarkers in 2016. The quantified transcripts of individuals (i.e. n = 10 from each of the four 2016 groups) were compared using two differential expression analysis methods (DESeq2 and edgeR), identifying a total of 1088 and 389 differentially expressed transcripts, respectively. Real-time quantitative polymerase chain reaction (qPCR) assays were designed and run for 83 of these transcripts to confirm RNA-seq results, identifying 9 transcripts with significantly higher expression after seismic exposure at the CC site, and 14 transcripts with significant differential expression between the CC site and the non-seismic LC site. Functional annotations associated with candidate seismic-responsive genes included oxidoreductase activity, apoptotic process, and inflammatory response. To further investigate the performance and utility of candidate biomarkers identified using the 2016 samples, expression levels of 20 selected transcripts were then examined in 2017 samples using both qPCR and multivariate statistical analyses. Principal component analysis of qPCR data separated exposure groups in 2016 but did not result in clear separation of groups in 2017, although some transcripts showed similar trends in both years. This work has built a strong foundation for future research involving the snow crab hepatopancreas transcriptome and its potential responses to environmental stressors including seismic surveying noise.

中文翻译：

---

雪蟹（Chionoecetes opilio）肝胰腺转录组：地震调查影响候选分子生物标志物的鉴定和测试

摘要 加拿大纽芬兰和拉布拉多 Grand Banks 的东坡是一个活跃的地震勘探区域，与重要的雪蟹 (Chionoecetes opilio) 渔业广泛重叠。2016 年和 2017 年进行了实地研究，以调查地震油气勘探暴露对雪蟹肝胰腺转录组的潜在影响，该转录组涉及新陈代谢和对环境压力的反应。2016 年，雪蟹在卡森峡谷 (CC) 暴露前（即震前）、暴露后 18 小时和 3 周进行 2 小时二维地震噪声采样。2017 年，重复进行二维地震暴露，并在 CC 暴露前和暴露后 1 天、2 天和 6 周收集样品。此外，在 2017 年，雪蟹经受 2 个月的 3D 地震噪声，并在暴露于 CC 以北的地点期间和暴露后 6 周进行采样。在这两年中，还从一个单独的非地震暴露地点 [Lilly Canyon (LC)] 收集了雪蟹。2016 年，RNA-seq 被用于鉴定候选的地震响应分子生物标志物。 使用两种差异表达分析方法（DESeq2 和 edgeR）比较个体（即 2016 年四个组中每组的 n = 10）的量化转录本，确定了一个总共分别有 1088 和 389 个差异表达的转录本。实时定量聚合酶链反应 (qPCR) 分析被设计并运行这些转录本中的 83 个以确认 RNA-seq 结果，在 CC 站点地震暴露后鉴定出具有显着更高表达的 9 个转录本，和 14 个在 CC 位点和非地震 LC 位点之间具有显着差异表达的转录本。与候选地震响应基因相关的功能注释包括氧化还原酶活性、细胞凋亡过程和炎症反应。为了进一步研究使用 2016 年样本确定的候选生物标志物的性能和效用，然后使用 qPCR 和多变量统计分析检查了 2017 年样本中 20 个选定转录本的表达水平。qPCR 数据的主成分分析在 2016 年分离了暴露组，但在 2017 年并未导致明确的组分离，尽管一些成绩单在这两年都显示出相似的趋势。