Laboratorio de Ecotoxicología Acuática, INIBIOMA (UNCo-CONICET) - CEAN, ruta provincial N° 61, Km 3, CCP 7, Junín de los Andes, 8371 Neuquén, Argentina.
Laboratorio de Ecotoxicología Acuática, INIBIOMA (UNCo-CONICET) - CEAN, ruta provincial N° 61, Km 3, CCP 7, Junín de los Andes, 8371 Neuquén, Argentina.
J Invertebr Pathol. 2018 Sep;157:36-44. doi: 10.1016/j.jip.2018.08.005. Epub 2018 Aug 9.
Immune cell characterization, immunological response and the associated gill oxidative balance were studied in the Patagonian freshwater mussel, Diplodon chilensis, using two microbiological immunostimulant models: Saccharomyces cerevisiae and Escherichia coli. Mussels were collected out of the breeding season in Paimún Lake and acclimated in the laboratory. Two exposure experiments were performed during two consecutive weeks: (1) mussels challenged with 500 yeast cells mL; and (2) mussels challenged with 1000 bacteria cells mL. Microorganisms were added in the water every two days, alternating with 6000 lyophilized cells of the green algae Scenedesmus vacuolatus mL. A control group, fed with S. vacuolatus, was set for each treatment. Morphological cell characterization was carried out in adherent hemocytes of D. chilensis hemolymph under control conditions. The most important cell type observed were the hyalinocytes (representing ca. 98% of the circulating cells), agranular cells with non-central polymorphic nucleus surrounded by cytoplasm; granulocytes (cells with cytoplasmic granules and non-central rounded nucleus) represented ca. 2%. Another two cell types were occasionally detected, binucleated hyalinocytes and hemoblast-like cells but were not considered for the analyses. Both adherent hyalinocytes and granulocytes exhibit phagocytic activity towards Congo red stained yeast, which was two-fold higher in granulocytes than in hyalinocytes, regardless of the applied challenge. Total hemocyte counts were diminished in mussels challenged with S. cerevisiae or E. coli. Hydrolytic and defense cellular enzyme activities were analyzed only for hyalinocytes. Both, S. cerevisiae and E. coli increased acid phosphatase activity. E. coli challenge diminished hemocyte lysosomal membrane stability and increased humoral phenoloxidase activity, while S. cerevisiae challenge did not affect any of these variables. Mussels challenged with E. coli showed increased gill antioxidant response without oxidative damage, while those challenged with S. cerevisiae showed no change in these variables.
采用两种微生物免疫刺激模型（酿酒酵母和大肠杆菌）研究了巴塔哥尼亚淡水贻贝 Diplodon chilensis 的免疫细胞特征、免疫反应和相关的鳃氧化平衡。贻贝在 Paimún 湖繁殖季节之外采集，并在实验室中适应。在连续两周内进行了两次暴露实验：（1）用 500 个酵母细胞 mL 挑战贻贝；（2）用 1000 个细菌细胞 mL 挑战贻贝。每隔两天向水中添加微生物，同时添加 6000 个冻干的绿藻 Scenedesmus vacuolatus 细胞 mL。为每个处理组设置了一个用 S. vacuolatus 喂养的对照组。在对照条件下，对 D. chilensis 血淋巴中的贴壁血红细胞进行形态细胞特征分析。观察到的最重要的细胞类型是透明细胞（约占循环细胞的 98%），无颗粒细胞，具有中央多形核，周围是细胞质；粒细胞（具有细胞质颗粒和中央圆形核的细胞）约占 2%。偶尔还检测到另外两种细胞类型，双核透明细胞和造血样细胞，但未纳入分析。贴壁透明细胞和粒细胞均对刚果红染色的酵母具有吞噬活性，粒细胞的吞噬活性是透明细胞的两倍，无论应用何种挑战都是如此。用 S. cerevisiae 或 E. coli 挑战贻贝会减少总血红细胞计数。仅对透明细胞分析水解和防御细胞酶活性。S. cerevisiae 和 E. coli 都增加了酸性磷酸酶活性。E. coli 挑战降低了血红细胞溶酶体膜稳定性并增加了体液酚氧化酶活性，而 S. cerevisiae 挑战对这些变量均无影响。用 E. coli 挑战的贻贝表现出增加的鳃抗氧化反应而没有氧化损伤，而用 S. cerevisiae 挑战的贻贝则没有这些变量的变化。
