Field validation of antioxidant enzyme biomarkers in mussels (Perna viridis) and clams (Ruditapes philippinarum) transplanted in Hong Kong coastal waters

Green-lipped mussels, Perna viridis, and Manila clams, Ruditapes philippinarum were sourced from “clean” sites in the Hong Kong region, depurated in a laboratory using uncontaminated filtered seawater for 8 days, and transplanted to a suspected gradient of chemically polluted sites in Hong Kong. After 14- and 28-days of field exposure, several antioxidant parameters including glutathione S transferase (GST), catalase (CAT), glutathione peroxidase (GPx), and glutathione (GSH) were quantified in gill and hepatopancreas tissues. Whole body tissue concentrations of polycyclic aromatic hydrocarbons (PAHs), petroleum hydrocarbons (PHCs), polychlorinated biphenyls (PCBs) and organochlorine pesticides (OCs) were determined in pooled site samples. Chemical analyses indicated that: (a) clams had higher levels of PAHs, PHCs, DDTs and PCBs, whereas mussels had higher hexachlorocyclohexane (HCHs) and there was no difference between species for dieldrin and remaining OCs; (b) Kat O should not be continued as a “clean” reference site for Hong Kong, because of the levels of contaminants measured and (c) PAH concentrations in the current survey were similar to those previously measured. Toxicological conclusions were: (a) antioxidant responses were different between species; (b) CAT and GST have highest utility in clams for field use in Hong Kong, whereas CAT in both gill and hepatopancreas tissue showed most potential in mussels; (c) significant induction of antioxidant responses over day 0 (excluding GPx in both tissues, and GST in mussel hepatic tissue); (d) groups of contaminants do not consistently induce antioxidant responses and (e) organochlorines and PCBs correlated significantly with CAT and GST in clam hepatopancreas and with CAT in mussel gill and hepatic tissue. Multivariate statistical techniques indicated little relationship between the site patterns for antioxidant responses and the contaminant gradients identified in body burden analysis.     

Okadaic acid, a causative toxin of diarrhetic shellfish poisoning, in green-lipped mussels Perna viridis from Hong Kong fish culture zones: Method development and monitoring

Green-lipped mussels (Perna viridis) were collected from seven fish culture zones (FCZs) in Hong Kong and analyzed for okadaic acid (OA). A conventional HPLC method was modified by incorporating a proteinase K digestion step. Results suggest that a higher recovery (2.5 times higher) of OA was obtained from spiked samples after the addition of 1.08 mg proteinase K in comparison with samples incubated without the proteolytic enzyme. For the hepatopancreas (HP) of individual field-collected mussels, the additional digestion step can enhance OA extraction by 3.1 times. Spatial and temporal variations in OA concentrations in the mussels from various FCZs were investigated. The highest concentration of OA in mussel HP samples was 1164.9 ng/g HP wet wt. With respect to OA concentrations in whole mussel tissues from seven sites and four seasonal samplings, the concentrations were between 70.0 and 131.0 ng/g wet wt., which did not exceed the generally recognized international regulatory criteria (>200 ng/g) for OA.     

Uptake and depuration of PAHs and chlorinated pesticides by semi-permeable membrane devices (SPMDs) and green-lipped mussels (Perna viridis)

Semi-batch seawater experiments were conducted to follow the uptake and release of selected PAHs (anthracene, fluorathene, pyrene and B[a]P) and organochlorine pesticides (-HCH, aldrin, dieldrin, p,p′-DDT) in semi-permeable membrane devices (SPMDs) and green-lipped mussels (Perna viridis). Mathematical models were applied to describe the uptake and elimination curves of the contaminants for SPMDs, and kinetic parameters, such as uptake rate constants, and equilibrium triolein/water partitioning coefficients were calculated. SPMD data showed a good fit to estimate rate constant and partition coefficient equations, but only those contaminants which partitioned mainly in the dissolved phase (-HCH and dieldrin) were well explained for mussels. Poor conformity of the other contaminants indicated mussels uptake by routes other than diffusion, such as ingestion of algae. An apparent equilibrium state was only noted for -HCH in mussels. Aldrin was not detected in mussels in the first few days of exposure, indicating potential metabolism of this compound. B[a]P was not detected in the triolein of SPMDs, which suggests that the membrane may act as a reservoir. Loss of spiked B[a]P from the triolein was evident in a depuration experiment, which may indicate transfer to the membrane. Rate constants for mussels were higher than those for SPMDs, but the reverse was true for partition coefficients. Overall, mussels and SPMDs had similar uptake rates for all compounds in this study, excluding p,p′-DDT and dieldrin. Contaminant elimination took place more rapidly in mussels, implying that SPMDs are better candidates for detecting episodic discharge of organic contaminants.     

泉州湾、围头湾翡翠贻贝海区半人工采苗技术研究

本文对泉州湾和围头湾翡翠贻贝的生殖腺性状、繁殖期、幼虫数量分布、附苗期及海区半人工采苗技术进行了研究。两湾翡翠贻贝繁殖期为5-9月,5月中旬至6月中旬为繁殖盛期,主要的附苗季节在6月中旬和7月上旬。选用胶胎绳和棕绳为采苗器,附苗效果较好;并讨论了投挂采苗器的适宜时间。     

温度对翡翠贻贝能量收支的影响

在不同的温度下对翡翠贻贝的摄食、耗氧、排氨、同化效率及能量收支等进行实验研究。结果表明，在设定的温度范围内，翡翠贻贝的摄食率、耗氧率、排氨率随温度的增加而增大，而效率则没有显著变化。在低温时，摄入的能量中分配于生长的比例较小，而代谢耗能和排泄耗能所占的比例相对较大，因而翡翠贻贝的生长较慢甚至出现负生长；而在高温时，翡翠贻贝由于用于生长的能量比例增加，而损失于代谢消耗和排泄的能量比例减少，因而生长预算和生长效率提高。     

东山湾翡翠贻贝海区半人工采苗技术研究

本文研究了东山湾翡翠贻贝的繁殖期，附苗期及海区半人工采苗技术。结果表明：东山湾翡翠贻贝的繁殖期在５－１０月，其５－６月为繁殖盛期。贻贝附苗季节在６－１１月，主要附苗季节在６月上中旬至７月初。通过对东山湾７片采苗区海水理化因子的测定和采苗效果的比较试验，确定了东山湾合适的采苗区为２＾＃－６＾＃区，其中３＾＃，４＾３，５＾＃区采苗效果最好，同时筛选出了木麻黄，胶皮合股绳和泡沫编织袋等３种附苗效果较好的     

Antioxidant responses to polycyclic aromatic hydrocarbons and organochlorine pesticides in green-lipped mussels (Perna viridis): Do mussels “integrate” biomarker responses?

Polycyclic aromatic hydrocarbons (PAHs) and organochlorine pesticides (OCs) are generally present in the marine environment in complex mixtures. The ecotoxicological nature of contaminant interactions, however, is poorly understood, with most scientific observations derived from single contaminant exposure experiments. The objective of this experiment was to examine dose-response relationships between antioxidant parameters and body contaminant levels in mussels exposed to different exposure regimes under laboratory conditions. Accordingly, the green-lipped mussel, Perna viridis, was challenged with a mixture of PAHs (anthracene, fluoranthene, pyrene, benzo[a]pyrene) and OC pesticides (α-HCH, aldrin, dieldrin, p,p′-DDT) over a 4 week period. Contaminants were delivered under four different dosing regimes, with all treatments receiving the same total contaminant load by the end of the exposure period. Antioxidant biomarkers were measured after 1, 2, 3 and 4 weeks, including glutathione (GSH), gluathione-S-transferase (GST), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR) and lipid peroxidase (LPO). GST and CAT were induced in hepatic tissues in most of the exposure regimes, with the majority of significant induction occurring in a constant exposure regime and a two-step alternate exposure regime. Significant differences among exposure regimes were detected in the body burden of contaminants after 28 days. Hepatic CAT and GSH are proposed as potentially useful biomarkers as they showed good correlation with target contaminants and were not readily affected by different dosing patterns.     

The usefulness of transplantation studies in monitoring of metals in the marine environment: South African experience

Due to their close proximity to the point sources of pollution, estuaries and harbours are exposed to metals. Mussels are used worldwide to monitor marine pollution due to their ability to take up contaminants and the ease of transplantation. Mussels were collected from two reference sites and transplanted in Richards Bay Harbour (2006 and 2010) and in three harbours. Transplanted and resident mussels were removed after 6 weeks exposure and metal concentrations were analysed using ICP-MS techniques. Transplanted mussels had higher metal concentrations than the resident mussels. This was attributed to regulation of metals by the resident mussels. Metal regulation was greater in mussels that are continuously submerged as opposed to those that undergo tidal influences. For monitoring it is essential that the reference population for transplantation is considered carefully as upwelling events in the pristine reference location results in significant increases in metal bioaccumulation.     

温度对2种饵料金藻保存效果的影响

研究了在室温25℃±2℃,4,-5,-22℃下绿色巴夫藻(Pavlova viridis)和球等鞭金藻(Isochrysis galbana)的保存效果,通过测定其存活率以及主要脂肪酸的变化情况筛选出了2种金藻的适宜保存温度。结果表明,绿色巴夫藻短期内(30~45 d)在-5℃下保存效果最好,较长期(45 d以上)保存则应在-22℃,而球等鞭金藻则在-22℃下保存效果最好。     

翡翠贻贝三个野生种群形态性状的差异分析

运用差异系数检验、主成分分析、判别分析和聚类分析等4种多元统计分析方法,采用5个比例性状,比较研究了广西北海、广东湛江和汕尾的3个翡翠贻贝野生种群的形态性状差异。结果表明:(1)3个种群间5个比例性状的差异系数在0.024～1.134之间,种群间尚未达到亚种差异水平;(2)主成分分析构建了3个主成分,PC1、PC2和PC3贡献率分别为40.134%、27.956%和20.614%,累计贡献率为88.704%;(3)利用逐步判别法选择3个贡献率较大的性状进行判别分析,建立了3种群判别函数,判别准确率为P1为73.0%～88.0%,P2为74.6%～86.8%,综合判别率为79.7%;(4)聚类分析显示,北海种群与汕尾种群形态较为接近,湛江种群趋异程度最大。各种分析方法从不同角度反映了群体间的形态学差异,这种差异是环境和遗传共同作用的结果。