Oxidative stress in two populations of killifish (Fundulus heteroclitus) with differing contaminant exposure histories

A population of killifish (Fundulus heteroclitus) inhabiting a creosote-polluted inlet of the Elizabeth River demonstrates tolerance to the acute toxic effects exerted by contaminated sediments on reference site killifish. Previous data have suggested that upregulated antioxidant defenses contribute to short-term tolerance in killifish exposed to Elizabeth River sediments. This study investigated population differences in antioxidant defenses from wild caught Elizabeth River and reference population killifish in different seasons, and after being held in the laboratory. Parameters measured in the killifish were total glutathione concentrations (GSH_T), activities of glutathione reductase (GR), glutathione peroxidase (GPx), and lipid peroxidation (LPO), all in adult hepatic tissues. The Elizabeth River population exhibited greater GSH_T, higher GPx activities, and increased LPO as compared to the reference population. Sex specific population differences were also observed in GSH_T and GPx. Both populations displayed decreased GSH_T and increased GR from early to late summer, as well as after being held in the laboratory. This study indicates that there are many factors that may contribute to differences in levels of antioxidant defenses in addition to exposure to contaminants, including reproductive status and environmental conditions.     

Patterns of heritability of decreased EROD activity and resistance to PCB 126-induced teratogenesis in laboratory-reared offspring of killifish (Fundulus heteroclitus) from a creosote-contaminated site in the Elizabeth River,VA, USA

Killifish (Fundulus heteroclitus) from a highly contaminated site on the Elizabeth River are resistant to the acute toxicity and the cytochrome P4501A (CYP1A)-inducing activity of both the sediments from the site and chemically pure polycyclic aromatic hydrocarbons (PAHs). These effects are highly heritable for one generation, but heritable to a lesser degree by subsequent generations, in clean conditions in the laboratory. We show that offspring of this population of Elizabeth River killifish are also resistant to the teratogenicity and P4501A-inducing activity of PCB congener 126, a prototypical coplanar halogenated aromatic hydrocarbon (HAH). Furthermore, the pattern of greater resistance to acute toxicity and P4501A-inducing activity in the first generation and less in subsequent generations is also observed upon exposure to PCB-126.     

Effects of contaminated sediments and sediment-exposed effluent water on an estuarine fish: Acute toxicity

The Elizabeth River is Virginia's most heavily populated, industrialized and contaminated subestuary. Its sediments contain the residues of wastes discharged for several hundred years. To examine some of the effects of sediment from a particularly noxious location, we chose the spot (Leiostomus xanthurus), a bottom-feeding sciaenid and naturally occurring spring-summer migrant of the river whose juveniles adapt readily to laboratory conditions. Spot from the Ware River, a nearby uncontaminated reference subestuary, were placed in two 380 liter (100 USgal), flow-through tanks (70 each). One tank contained Elizabeth River sediments contaminated with PAHs. The second tank contained uncontaminated sediment from the York River as a control. 13 spot each were placed in two aquaria that received the overflow from the sediment-containing tanks. Animals from the reference estuary were examined and processed to provide baseline information.Results obtained to date show the following: (1) spot experimentally exposed to contaminated sediment developed penetrating integumental lesions within 8 days after exposure began and later severe fin and gill erosion; (2) their hematocrits were significantly reduced and no weight gain occurred; (3) pancreatic and liver alterations were observed in some of the chemically stressed fish; (4) control fish exhibited no fin erosion or integumental lesions; (5) control fish showed no hematocrit or growth reduction; and (6) dead fish were first observed in the contaminated-sediment tank after 8 days while no control fish died. Clearly, one or more factors associated with Elizabeth River sediments and/or water closely associated with those sediments (probably the heavy concentrations of PAHs) are capable of causing serious injury, disease and death in experimental spot populations.     

海水酸化和磺胺甲恶唑复合暴露对大马蹄螺抗氧化酶活性和脂质过氧化的影响

为探讨海水酸化和磺胺甲恶唑环境残留对珊瑚礁区生物的影响,本文以南海珊瑚礁区关键礁栖生物大马蹄螺（Trochus niloticus） 为研究对象,探究了两种环境胁迫因素对大马蹄螺的氧化胁迫作用。分别采用两个 pH 水平 （pH 8.1、 pH 7.6） 和三个磺胺甲恶唑浓度水平 （0 ng/L、100 ng/L 和 1 000 ng/L） 单独和复合暴露 28 天,测定其对大马蹄螺鳃组织的超氧化物歧化酶 （SOD）、过氧化氢酶 （CAT） 和谷胱甘肽巯基转移酶 （GST） 活性以及谷胱甘肽 （GSH） 含量和脂质过氧化水平 （LPO） 的影响。结果发现：海水酸化和磺胺甲恶唑单独暴露能够导致大马蹄螺鳃组织出现不同程度的氧化应激现象,表现为抗氧化酶 （CAT、SOD 和 GST） 活性和谷胱甘肽含量的升高。在复合暴露条件下,大马蹄螺鳃组织的抗氧化酶（CAT、SOD 和 GST） 活性和谷胱甘肽含量均受到不同程度的抑制,同时伴随着脂质过氧化水平的显著升高。这表明短期海水酸化和磺胺甲恶唑复合暴露可导致大马蹄螺鳃组织出现氧化损伤现象,进而可能对个体存活和种群繁衍造成负面影响。     

基于氧化逆境标志物的北部湾潮间带沉积物有机污染程度综合评价

利用综合生物标志物响应(IBR)指数对北部湾潮间带沉积环境中多氯联苯(PCBs)、石油烃(TPH)、壬基酚(NP) 和苯并芘(B[a]P)的综合污染水平进行评价。在9个站位采集波纹巴非蛤(Paphia undulata)和沉积物样品,测定沉积物和生物软组织中有机污染物含量,同时对鳃、内脏中与细胞防御系统有关的各7种标志物(包括还原型谷胱甘肽(GSH)、氧化型谷胱甘肽(GSSG)、谷胱甘肽过氧化物酶(GPx)、过氧化氢酶(CAT)等)进行分析。利用相关性分析法筛选出对1~2种有机污染物响应敏感的生物标志物共8种,分别为内脏中的GPx、GSH、GSSG、CAT和鳃中的GPx、GSH、GSSG、CAT。以筛选的8种生物标志物为基础, 9个站位IBR指数范围为0.27~1.48,其中站位S2(湛江市乐民镇码头)、站位S9(湛江市江洪码头)的IBR较小,显示污染程度最轻;站位S4(防城港西湾)、站位S3(钦州湾老人沙浅滩) IBR较高,显示生物体因有机污染产生氧化逆境。不同站位的IBR值与污染源分布、化学污染物分析结果基本一致,表明该方法可用于北部湾潮间带沉积物有机污染程度的综合评价。     

Seasonal variation of oxidative biomarkers in gills and digestive gland of green-lipped mussel Perna viridis from Arabian Sea

Investigations on seasonal variation in oxidative stress biomarkers were carried out on the natural population of green-lipped mussel Perna viridis collected from Bambolim beach area of Goa. Oxidative stress indices such as lipid peroxidation (LPX), hydrogen peroxide (H₂O₂), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX), glutathione transferase (GST), glutathione reductase (GR), reduced glutathione (GSH) and ascorbic acid (ASA) were measured in gills and digestive gland of P. viridis during February, May, August and November. The present study reveals two important aspects regarding the antioxidant defence status of tissues of P. viridis. Firstly, antioxidant capacity of tissues of P. viridis exhibits seasonal variation. Secondly, various components of antioxidant capacity such as oxidative stress markers, levels of antioxidant enzymes and small antioxidant molecules vary differently in tissues with respect to different seasons. Although the oxidative stress status of gills and digestive gland of P. viridis expressed in terms of LPX and H₂O₂ was the lowest in February, its level was maximal in gills and digestive gland during May and November, respectively. While activities of SOD and GPX of tissues of P. viridis were found to be low in August, activities of CAT and GR were recorded to be low in February. GST activity in gills although remained high in February, in digestive gland elevated values were recorded in August and November. A seasonal variation in the levels of small antioxidant molecules was also noticed. Among non enzymatic antioxidants ASA content of tissues was maximal in May and August in comparison to February and November, but GSH remained high in November. It therefore appears that environmental factors may play a crucial role in regulating the oxidative stress capacity of tissues of P. viridis.     

丙溴磷对二种海洋微藻的GPx活性及GSH、CAR含量的影响

实验以三角褐指藻和青岛大扁藻为材料 ,进行了丙溴磷农药对海洋微藻的抗氧化防御系统成份— GPx(glutatione Peroxidase)活性和 GSH(glutathione)、CAR(carotinoid)含量的影响研究 ,结果表明 ,在丙溴磷胁迫下 ,微藻的 GPx活性呈现下降趋势 ,GSH和 CAR含量也表现为下降趋势 ,并且胁迫的时间越长、胁迫的强度越大它们下降的幅度也越大。因此微藻细胞对抗氧化防御系统的能力被削弱 ,从而可能导致活性氧在体内的过量产生与积累 ,进而对藻细胞造成更大的伤害。     

Temporal variation in the antioxidant defence system and lipid peroxidation in the gills and mantle of hydrothermal vent mussel Bathymodiolus azoricus

Hydrothermal vent mussels are exposed continually to toxic compounds, including high metal concentrations and other substances like dissolved sulphide, methane and natural radioactivity. Fluctuations in these parameters appear to be common because of the characteristic instability of the hydrothermal environment. Temporal variation in the antioxidant enzymes (superoxide dismutase (SOD), catalase (CAT), total glutathione peroxidases (Total GPx), selenium dependent glutathione peroxidases (Se-GPx)), metallothioneins and lipid peroxidation (LPO) in the gills and mantle of the mussel Bathymodiolus azoricus from Menez–Gwen hydrothermal vent site was evaluated and related to the accumulated metal concentrations (Ag, Cu, Cd, Fe, Mn and Zn) in the tissues. Maximum antioxidant enzyme activities in the gills were detected in the beginning of summer, followed by a gradual decrease throughout the following months. One year after, the levels of antioxidant enzyme activities were similar to those reported one year before. LPO in this tissue exhibited a similar temporal variation trend. A different pattern of temporal variation in antioxidant enzyme activities was observed in the mantle, with a gradual increase from summer to the end of autumn (November). LPO in the mantle exhibited an almost reverse trend of temporal variation to that of antioxidant enzyme activities in this tissue. Antioxidant defences in the gills of B. azoricus were significantly enhanced with increasing concentrations of Ag, Cu and Mn, while negative relationships between antioxidant enzymes and Cd, Cu, Mn and Zn concentrations in the mantle were observed, suggesting different pathways of reactive oxygen species (ROS) production and that these tissues responded differently to the metal accumulation. However, temporal variation in biomarkers of defence and damage were in general similar to coastal bivalve species and can be associated with temporal variations of the physiological status due to reproduction. These variations might also be linked to the highly unstable nature of the hydrothermal environment.     

Preliminary investigations of the chemiluminescent response in normal and pollutant-exposed fish

A luminol-dependent chemiluminescence (CL) assay is currently being developed and tested with macrophages isolated from the kidneys of spot (Leiostomus xanthrus) and hogchoker (Trinectes maculatus). Both peak amplitude and time of maximum response have been observed to vary with the species tested and the stimulus used. In York River (reference), spot zymosan elicited the most immediate response with a peak at 18 min, while the response induced by Staphylococcus aureus was delayed (28 min). In York River (reference) hogchoker, the most immediate response was induced by phorbol myristate acetate (PMA) with a peak at 7min. S. aureus and zymosan elicited somewhat delayed responses (13 min and 15 min, respectively). Macrophages from spot captured in the Elizabeth River, which is highly contaminated with polynuclear aromatic hydrocarbons (PAHs), were markedly deficient in generating a zymosan-induced CL response compared with York River reference fish. Spot exposed in the laboratory to contaminated sediments showed a similar decrease in the CL response.     

Effects of toxic chemicals on macrophage phagocytosis in two estuarine fishes

The phagocytic efficiency of macrophages isolated from the kidney of spot, Leiostomus xanthurus, and hogchoker, Trinectes maculatus, was determined in fish captured from the Elizabeth River (Virginia) Macrophage phagocytosis was found to be markedly reduced in the Elizabeth River fish as compared to Ware River controls. The macrophage phagocytic activity of Elizabeth River fish returned to normal after the fish were held in clean water for several weeks. This indicates that the decreased phagocytic activity was related to exposure to Elizabeth River pollutants but may be reversible.     

军曹鱼（Rachycentron canadum）幼鱼对环境低氧胁迫氧化应激与能量利用指标的响应

本文探究环境低氧对军曹鱼(Rachycentron canadum)氧化应激和能量利用指标的影响,为军曹鱼的健康养殖提供参考依据。通过设置低氧胁迫–恢复实验,将军曹鱼幼鱼(平均体质量(220.67±20.73)g)在低氧((2.64±0.25)mg/L)胁迫3 h及复氧((6.34±0.15)mg/L)8 h、24 h和48 h后,测定其肝脏和肌肉组织的氧化应激与能量利用指标。结果显示,低氧胁迫后,肝脏中丙二醛（Malondialdehyde,MDA)、过氧化氢酶(Catalase,CAT)和谷胱甘肽还原酶(Glutathione Reductase,GR)活力均显著低于对照组(p<0.05),乳酸脱氢酶(Lactate Dehydrogenase,LDH)活性显著高于对照组(p <0.05);肌肉中MDA和脂质过氧化物(Lipid Peroxidase,LPO)活性均显著低于对照组(p<0.05),超氧化物歧化酶(Superoxide Dismutase,SOD)和LDH活性均显著高于对照组(p<0.05);肌糖原和肝糖原含量极显著低于对照组(p<0.01)。复氧过程中,肝脏和肌肉中MDA、LPO、SOD、CAT、谷胱甘肽过氧化物酶(Glutathione Peroxidase,GPx)和GR含量均出现不同程度的升高;肝糖原在复氧24 h后显著高于对照组(p<0.05),复氧48 h后显著低于对照组(p<0.05);肌糖原在复氧8 h、24 h和48 h后均显著低于对照组(p<0.05)。研究表明,低氧胁迫能够对军曹鱼幼鱼机体造成一定的氧化损伤,肝脏和肌肉组织的酶活力和能量供应发生变化;低氧胁迫后的再复氧环境,对机体造成更为强烈的氧化损伤,可通过自身生理调节逐渐恢复到正常水平。     

Teratogenesis in Fundulus heteroclitus embryos exposed to a creosote-contaminated sediment extract and CYP1A inhibitors

The goal of these experiments was to explore the relationship between cytochrome P4501A (CYP1A) induction and the teratogenicity of sediments from the Atlantic Wood Industries Superfund site (Elizabeth River, VA) in Fundulus heteroclitus embryos. In these experiments we used embryos spawned from reference site adults to assess CYP1A activity and teratogenicity induced by aqueous Elizabeth River sediment extracts (ERSE). Embryo exposures to ERSE induced CYP1A activity and caused deformities, including pericardial edema, heart elongation and tail shortening. Co-exposures with various CYP1A inhibitors significantly decreased CYP1A activity and increased the teratogenicity of the sediment extract. Potential mechanisms for this increased toxicity are discussed herein.     

Adaptation of the antioxidant defence system in hydrothermal-vent mussels (Bathymodiolus azoricus) transplanted between two Mid-Atlantic Ridge sites

The vent mussel Bathymodiolus azoricus is the dominant member of the Northern Mid‐Atlantic Ridge (MAR) hydrothermal megafauna, and lives in an environment characterized by temporal and spatial variations in the levels of heavy metals, methane and hydrogen sulphide, substances which are known to increase reactive oxygen species levels in the tissues of exposed organisms. To evaluate the effects of two contrasting hydrothermal environments on the antioxidant defence system of this vent mussel species, a 2‐week transplant experiment was carried out involving mussels collected from the relatively deep (2300 m), and chemical rich, Rainbow vent field. These were transplanted to the shallower (1700 m), and relatively less toxic, Lucky Strike vent field. To achieve this objective, levels of superoxide dismutase, catalase (CAT), total glutathione peroxidase (GPx), selenium‐dependent glutathione peroxidase and lipid peroxidation (LPO) were measured in the gills and mantle tissues of resident and transplant mussels before and after the transplant experiment. With the exception of CAT, the gills of the transplanted mussels had significantly higher antioxidant enzyme activity compared with the basal levels in the donor (Rainbow) and recipient (Lucky Strike) populations; whereas the antioxidant enzyme levels in the mantle tissues of the transplants reflected the baseline levels of activity in the native Lucky Strike mussels after 2 weeks. In contrast, LPO levels were significantly higher in both tissue types in the transplants than in either the source or the recipient populations, which suggested a response to hydrostatic pressure change (note, the transplant animals were brought to the surface for transportation between the two vent fields). The fact that the Rainbow mussels survived the transplant experience indicates that B. azoricus has a very robust constitution, which enables it to cope behaviourally, physiologically and genetically with the extreme conditions found in its naturally contaminated deep‐sea environment.     

Protein carbonyls and antioxidant defenses in corkwing wrasse (Symphodus melops) from a heavy metal polluted and a PAH polluted site

The use of fish in environmental monitoring has become increasingly important in recent years as anthropogenic substances, many of which function as prooxidants, are accumulating in aquatic environments. We have measured a battery of antioxidant defenses as a measure of oxidative status, as well as protein carbonylation as a measure of oxidative damage, in corkwing wrasse (Symphodus melops) captured near a disused copper mine, where water and sediment are contaminated with heavy metals, and an aluminum smelter, a site contaminated with PAHs. Results were compared to two different reference sites. Fish at the heavy metal site had lower glucose-6-phosphate dehydrogenase activity and elevated protein carbonyls (1.8 times) compared to fish from the reference site. At the PAH site, EROD was increased 2-fold, while total glutathione and methemoglobin reductase concentration, were decreased. No differences were seen in protein carbonyl levels at the PAH site. Measures of both antioxidant defenses and oxidative damage should be used when assessing effects of xenobiotics on oxidative stress in fish species.     

Teratogenesis in Fundulus heteroclitus embryos exposed to a creosote-contaminated sediment extract and CYP1A inhibitors

The goal of these experiments was to explore the relationship between cytochrome P4501A (CYP1A) induction and the teratogenicity of sediments from the Atlantic Wood Industries Superfund site (Elizabeth River, VA) in Fundulus heteroclitus embryos. In these experiments we used embryos spawned from reference site adults to assess CYP1A activity and teratogenicity induced by aqueous Elizabeth River sediment extracts (ERSE). Embryo exposures to ERSE induced CYP1A activity and caused deformities, including pericardial edema, heart elongation and tail shortening. Co-exposures with various CYP1A inhibitors significantly decreased CYP1A activity and increased the teratogenicity of the sediment extract. Potential mechanisms for this increased toxicity are discussed herein.     

中华哲水蚤抗氧化系统对海水酸化的响应

海洋浮游动物可以对因二氧化碳浓度升高导致的海水酸化做出敏感的响应,然而关于海水酸化对其生理机能的研究却较少。因此,我们研究了暴露于不同二氧化碳浓度（0.08,0.20,0.50 and 1.00%）的酸化条件下中华哲水蚤主要抗氧化酶以及2种解毒酶的活性变化情况。结果表明,暴露于酸化海水中的中华哲水蚤的GPx酶活性显著高于对照组,然而,其它抗氧化组分,包括GST、SOD活性,GSH水平和GSH/GSSG比值均被显著抑制。中华哲水蚤的ATPase活性被海水酸化显著刺激,而AchE活性却显著被抑制。此外,主成分分析结果表明,各指标变化的75.93%可以用第一和第二主成分解释。因二氧化碳浓度升高导致的海水酸化可以通过影响桡足类体内一些酶类的活性来影响其新陈代谢及存活。在未来的研究工作中,需要对海水酸化和其它环境因素的协同作用对桡足类的影响作进一步研究。     

卵磷脂对花尾胡椒401幼鱼谷肤甘肤一硫一转移酶，谷肤甘肤过氧化物酶和DNA完整性的影响

采用不同卵磷脂添加量的配合饲料投喂花尾胡椒鲷(Plectorhyncchus cinctus）幼鱼，同步测定幼鱼的谷胱甘肽-硫-转移酶和谷胱甘肽过氧化物酶活性以及DNA完整程度。结果表明，谷胱甘肽-硫-转移酶和谷胱甘肽过氧化物酶活性随着饲料中卵磷脂含量的增加而提高，而DNA单链断裂程度随着饲料中卵磷脂含量的增加而减少。这提示卵磷脂能够提高鱼体的抗氧化防御系统的机能。     

Seasonal changes of antioxidant and oxidative parameters in the coral Pocillopora capitata on the Pacific coast of Mexico

The physiological responses of the coral Pocillopora capitata to environmental conditions common in winter and summer were studied in 2007 during February–March (winter) and June–July (summer) at La Boquita reef (Manzanillo, Colima, Mexico). Shallow and deep sampling stations were established at different distances from a small jetty built next to the Juluapan Lagoon. We analyzed superoxide radicals () and lipid peroxidants (TBARS); the activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR), and glutathione‐S‐transferase (GST); chlorophyll a (Chl a), zooxanthellae density (ZD); and mycosporine‐like amino acids (MAAs). Our results showed that the , TBARS, CAT, GST, MAAs, and Chl a, levels were significantly higher in summer (P < 0.05); no seasonal difference was found for GPx, GR or ZD. We found significant differences (P < 0.05) in winter only for Chl a and ZD at shallow sites and, in contrast, for at deeper sites. The results of this study indicate that increasing temperature and radiation associated with seasonal changes (from winter to summer), the efficiency of the enzymes GST, CR and GPX, and the production of MAAs together form a powerful mechanism for P. capitata to offset the detrimental effects of environmental change.     

Effect of cadmium, copper and mercury on antioxidant enzyme activities and lipid peroxidation in the gills of the hydrothermal vent mussel Bathymodiolus azoricus

Metals are known to influence lipid peroxidation and oxidative status of marine organisms. Hydrothermal vent mussels Bathymodiolus azoricus live in deep-sea environments with anomalous conditions, including high metal concentrations. Although B. azoricus are aerobic organisms they possess abundant methano and thioautotrophic symbiotic bacteria in the gills. The enzymatic defences (superoxide dismutase (SOD), catalase (CAT), total glutathione peroxidase (Total GPx) and selenium-dependent glutathione peroxidase (Se–GPx)) and lipid peroxidation were determined in the gills of B. azoricus exposed to Cd (0.9 μM), Cu (0.4 μM) and Hg (0.1 μM) with different times of exposure. The experiments were performed in pressurized containers at 9 ± 1 °C and 85 bars.Results show that vent mussels possess antioxidant enzymatic protection in the gills. Cd and Cu had an inhibitory effect in the enzymatic defence system, contrarily to Hg. These enzymatic systems are not completely understood in the B. azoricus, since reactive oxygen species might be produced through other processes than natural redox cycling, due to hydrogen sulphide and oxygen content present. Also the symbiotic bacteria may play an important contribution in the antioxidant protection of the gills.     

氟苯尼考对菲律宾蛤仔抗氧化酶活性的影响

通过室内模拟实验研究了氟苯尼考对菲律宾蛤仔体内超氧化物歧化酶(SOD),过氧化氢酶(CAT),过氧化物酶(POD)和谷胱甘肽还原酶(GR)活性的影响.结果表明,培养7 d后,在氟苯尼考的作用下,菲律宾蛤仔内脏团和肌肉组织的SOD,CAT,POD和GR的活性都发生了变化.低质量比氟苯尼考对SOD,CAT和POD酶活性具有诱导作用,而在高质量比条件下则表现为抑制作用.不同质量比的氟苯尼考对菲律宾蛤仔内脏团和肌肉组织中的GR活性都有明显的抑制作用,且随着氟苯尼考质量比的增加抑制作用加强.