Dorantes-Aranda Juan José, Nichols Peter D, David Waite Trevor, Hallegraeff Gustaaf M
Institute for Marine and Antarctic Studies, University of Tasmania, Private Bag 55, Hobart, Tasmania, 7001, Australia.
CSIRO Wealth from Oceans Flagship, Marine and Atmospheric Research, GPO Box 1538, Hobart, Tasmania, 7001, Australia.
J Phycol. 2013 Apr;49(2):427-38. doi: 10.1111/jpy.12053. Epub 2013 Mar 15.
Lipid profiles of three strains (Mexico, Australia, Japan) of Chattonella marina (Subrahmanyan) Hara et Chihara were studied under defined growth (phosphate, light, and growth phase) and harvest (intact and ruptured cells) conditions. Triacylglycerol levels were always <2%, sterols <7%, free fatty acids varied between 2 and 33%, and polar lipids were the most abundant lipid class (>51% of total lipids). The major fatty acids in C. marina were palmitic (16:0), eicosapentaenoic (EPA, 20:5ω3), octadecatetraenoic (18:4ω3), myristic (14:0), and palmitoleic (16:1ω7c) acids. Higher levels of EPA were found in ruptured cells (21.4-29.4%) compared to intact cells (8.5-25.3%). In general, Japanese N-118 C. marina was the highest producer of EPA (14.3-29.4%), and Mexican CMCV-1 the lowest producer (7.9-27.1%). Algal cultures, free fatty acids from C. marina, and the two aldehydes 2E,4E-decadienal and 2E,4E-heptadienal (suspected fatty acid-derived products) were tested against the rainbow trout fish gill cell line RTgill-W1. The configuration of fatty acids plays an important role in ichthyotoxicity. Free fatty acid fractions, obtained by base saponification of total lipids from C. marina showed a potent toxicity toward gill cells (median lethal concentration, LC50 (at 1 h) of 0.44 μg · mL(-1) in light conditions, with a complete loss of viability at >3.2 μg · mL(-1) ). Live cultures of Mexican C. marina were less toxic than Japanese and Australian strains. This difference could be related to differing EPA content, superoxide anion production, and cell fragility. The aldehydes 2E,4E-decadienal and 2E,4E-heptadienal also showed high impact on gill cell viability, with LC50 (at 1 h) of 0.34 and 0.36 μg · mL(-1) , respectively. Superoxide anion production was highest in Australian strain CMPL01, followed by Japanese N-118 and Mexican CMCV-1 strains. Ruptured cells showed higher production of superoxide anion compared to intact cells (e.g., 19 vs. 9.5 pmol · cell(-1) · hr(-1) for CMPL01, respectively). Our results indicate that C. marina is more ichthyotoxic after cell disruption and when switching from dark to light conditions, possibly associated with a higher production of superoxide anion and EPA, which may be quickly oxidized to produce more toxic derivates, such as aldehydes.
在特定的生长条件(磷酸盐、光照和生长阶段)及收获条件(完整细胞和破裂细胞)下,对哈氏赤潮藻(Chattonella marina (Subrahmanyan) Hara et Chihara)的三个菌株(墨西哥、澳大利亚、日本)的脂质概况进行了研究。三酰甘油水平始终<2%,甾醇<7%,游离脂肪酸在2%至33%之间变化,极性脂质是最丰富的脂质类别(占总脂质的>51%)。哈氏赤潮藻中的主要脂肪酸为棕榈酸(16:0)、二十碳五烯酸(EPA,20:5ω3)、十八碳四烯酸(18:4ω3)、肉豆蔻酸(14:0)和棕榈油酸(16:1ω7c)。与完整细胞(8.5 - 25.3%)相比,破裂细胞中EPA水平更高(21.4 - 29.4%)。总体而言,日本的N - 118哈氏赤潮藻是EPA的最高生产者(14.3 - 29.4%),墨西哥的CMCV - 1是最低生产者(7.9 - 27.1%)。对虹鳟鱼鳃细胞系RTgill - W1测试了藻类培养物、哈氏赤潮藻的游离脂肪酸以及两种醛类物质2E,4E - 癸二烯醛和2E,4E - 庚二烯醛(疑似脂肪酸衍生产物)。脂肪酸的构型在对鱼类的毒性方面起着重要作用。通过对哈氏赤潮藻总脂质进行碱皂化获得的游离脂肪酸部分对鳃细胞显示出强大的毒性(在光照条件下,1小时的半数致死浓度LC50为0.44μg·mL⁻¹,在>3.2μg·mL⁻¹时活力完全丧失)。墨西哥哈氏赤潮藻的活培养物毒性低于日本和澳大利亚菌株。这种差异可能与EPA含量、超氧阴离子产生以及细胞脆性的不同有关。醛类物质2E,4E - 癸二烯醛和2E,4E - 庚二烯醛对鳃细胞活力也有很大影响,其1小时的LC⁵₀分别为0.34和0.36μg·mL⁻¹。超氧阴离子产生量在澳大利亚菌株CMPL01中最高,其次是日本的N - 118和墨西哥的CMCV - 1菌株。与完整细胞相比,破裂细胞显示出更高的超氧阴离子产生量(例如,CMPL01的完整细胞和破裂细胞分别为9.5和19 pmol·cell⁻¹·hr⁻¹)。我们的结果表明,哈氏赤潮藻在细胞破裂后以及从黑暗转为光照条件时对鱼类的毒性更强,这可能与超氧阴离子和EPA的产生增加有关,EPA可能会迅速氧化生成毒性更强的衍生物,如醛类物质。