The effects of ocean acidification on hemocyte of crab species in Alaska from laboratory experiment studies from 2011-07-01 to 2013-07-06 (NCEI Accession 0123400)

Flow cytometry provides a rapid and reproducible method for analyzing crustacean hemocytes and their functions under experimentally-varied environmental conditions. We used flow cytometry to determine if there was a difference in hematology and selected immune functions, and intracellular pH (pHi), under two different, future ocean acidification scenarios (pH = 7.51, 7.80) compared to current conditions (pH = 8.06) for Chionoecetes bairdi, the Tanner crab. Hemocytes were analyzed after adult Tanner crabs were held for two years under continuous exposure to acidified ocean water. Total counts of hemocytes did not vary among experimental control and treatments; however, there was a significantly greater number of dead, circulating hemocytes in crabs held at the lowest pH treatment. Phagocytosis of fluorescent microbeads by hemocytes was greatest at the lowest pH treatment. These results suggest that hemocytes were dying, likely by apoptosis, at a rate faster than upregulated phagocytosis was able to remove moribund cells from circulation at the lowest pH. There was no significant difference in pHi within hyalinocytes among pH levels, with apparent regulation to a mean pHi of 7.24, significantly lower than the external environment. In contrast, there was a significant difference between treatments in pHi of the semi-granular+granular cells. These findings suggest that future, predicted levels of ocean acidification may affect the defense cells of Tanner crabs, possibly making them more susceptible to other stressors in the environment.

流式细胞术（Flow cytometry）为分析甲壳类血细胞及其在实验性改变环境条件下的功能提供了一种快速且可重复的方法。我们采用流式细胞术，针对 Tanner蟹（学名Chionoecetes bairdi），对比当前环境条件（pH=8.06）与两种不同的未来海洋酸化情景（pH=7.51、7.80），探究其血液学特征、选定免疫功能及细胞内pH值（pHi）是否存在差异。成年 Tanner蟹在持续暴露于酸化海水环境两年后，我们对其血细胞进行了分析。血细胞总数在实验对照组与各处理组间无显著差异；然而，在最低pH处理组中，蟹体内死亡的循环血细胞数量显著增加。血细胞对荧光微球的吞噬作用在最低pH处理组中最强。这些结果表明，在最低pH条件下，血细胞死亡速率（可能通过细胞凋亡（apoptosis））快于上调的吞噬作用清除循环中濒死细胞的速率。不同pH水平下，透明细胞内的pHi无显著差异，其平均pHi被明显调节至7.24，显著低于外部环境pH值。相比之下，半颗粒+颗粒细胞的pHi在各处理组间存在显著差异。这些发现表明，未来预测的海洋酸化水平可能影响 Tanner蟹的防御细胞，使其可能更易受环境中其他应激源的影响。