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.     

The DNA de-methylating agent 5-azacytidine does not restore CYP1A induction in PCB resistant Newark Bay killifish (Fundulus heteroclitus)

Newark Bay (NB) killifish (Fundulus heteroclitus) have been chronically exposed to environmental contaminants that activate the aryl hydrocarbon receptor (AHR) and are tolerant to toxic effects and CYP1A induction provoked by AHR ligands. Resistance to CYP1A induction could be due to an epigenetic mechanism such as DNA methylation. We measured in-ovo CYP1A catalytic activity (ethoxyresorufin-O-deethylase, EROD) in NB and reference site killifish embryos aqueously exposed to various concentrations of the de-methylating agent 5-azacytidine, 5-AC (5, 50 and 500 μ(micro)M) with or without 0.2 μ(micro)g/l of the CYP1A inducer 3,3^′,4,4^′,5 pentachlorobiphenyl (IUPAC PCB126). Neither PCB126 alone, nor PCB126 plus 5-AC, induced EROD above levels in vehicle treated Newark Bay fish. In reference site fish, the same PCB126 dose provoked a 7.4-fold EROD induction relative to controls. We conclude that Newark Bay killifish are resistant to CYP1A induction by co-planar PCBs during early embryological development and our data suggests that DNA methylation does not play a critical role in resistance to CYP1A induction in this model.     

Life-history patterns of the salt-marsh killifish Fundulus heteroclitus (L.) introduced in the estuary of the guadalquivir river (South West Spain)

Two thousand four hundred and eighty-four specimens from a population of Fundulus heteroclitus introduced into the southwestern Iberian Peninsula were used to carry out a study on age, growth and reproduction. In this new habitat the species displayed the same life-history tactics as in its original areas. The life-span was shorter, with a winter age-structure of 67·4% in 0 + group, 29·1% in 1 + group, 2·5% in 2 + group and 1·1% in 3 + group. Females showed a higher growth rate and a longer growth period than males and because of this they were always longer than males in all the age-groups. In both sexes growth stopped and the somatic condition was at a minimum from April to June, when the gonadosomatic index reached its highest values. The maximum average gonadosomatic index in females was four times higher than that of males. Specimens achieved maturity during the spawning season the year following birth at a minimum average total length of 45 mm for males and 60 mm for females. Fecundity was related to fish weight, total length and gonad weight. The overall sex ratio did not differ significantly from 1:1.     

Relevance of intracellular partitioning of metals in prey to differential metal bioaccumulation among populations of mummichogs (Fundulus heteroclitus)

Intracellular partitioning of trace metals is critical to metal tolerance in aquatic organisms and may also influence metal trophic transfer in ecosystems. In this study, we tested the relevance of metal (Cd, Cu, Pb, and Zn) intracellular partitioning in prey as an indicator of metal trophic availability to benthic forage fish, mummichogs (Fundulus heteroclitus), in chronically metal-polluted salt marshes in New York, USA. Two common prey of mummichogs in the study area, Palaemonetes pugio and Nereis acuminata, generally stored increasingly higher proportions of non-essential metals (particularly Pb) in insoluble (less trophically available) cellular components, as the whole body burdens increased. In contrast, intracellular partitioning of essential metals (Cu and Zn) in invertebrate prey varied relatively little among sites. Differential Cd and Pb intracellular partitioning patterns within P. pugio among sites were significantly associated with Cd and Pb whole body burdens in mummichogs, respectively (i.e., prey-driven bioreduction of metals), while bioaccumulation of Cu and Zn in mummichogs was similar among populations. The findings in this study suggest that metal intracellular partitioning within prey may be partially responsible for metal trophic availability to a predator in metal-polluted habitats, while there was also evidence that some predator-dependent processes may offset differential trophic availabilities from prey.     

Hepatic histopathology in two populations of the mummichog, Fundulus heteroclitus

The pollution-tolerant mummichog, Fundulus heteroclitus, inhabits a heavily contaminated estuary, Piles Creek (PC), in Linden, NJ. PC contaminants include PAHs, PCBs and many metals, including high Hg and Pb. PC mummichog livers and kidneys were compared with those from a pristine estuary in Tuckerton (TK), NJ, by standard histopathological methods, with quantification by computerized image analysis. PC livers had>2× the lipid ratios of TK livers. Cell sizes and shapes were not different between populations. TK livers had a 25% incidence of basophilic areas and an age-related 30% incidence of macrophage aggregates; PC livers had none of these lesions. PC livers had a 100% incidence of necrotic foci, compared with 55% of TK livers. Inflammation was noted in 50% of TK livers but only 27% of PC livers. Curiously, the TK livers had a 45% incidence of metacercarial cysts of a digenetic trematode, while none were found in the PC livers. The TK focal necrosis and macrophage aggregates coincided with parasite infestation. There were no observed kidney lesions in either population. Positive biomarkers in this species thus included high hepatic lipid content, possibly necrosis, but no other lesions. The lack of trematode cysts in PC livers may reflect the lack of an intermediate host in this low biodiversity estuary.     

Response of a benthic suspension feeder (Crassostrea virginica Gmelin) to three centuries of anthropogenic eutrophication in Chesapeake Bay

Biogenic reefs built by oysters and other suspension feeders are vital components of estuarine ecosystems. By consuming phytoplankton, suspension feeders act to suppress accumulation of organic matter in the water column. Nutrient loading increases the rate of primary production, thereby causing eutrophication. As suspension feeders consume more organic matter from increasing abundance of phytoplankton, their rate of growth should also increase if they are food limited. We show here that the eastern oyster, Crassostrea virginica (Gmelin), from St. Mary's and Patuxent rivers, Chesapeake Bay, grew faster during anthropogenic eutrophication relative to C. virginica before eutrophication. Growth of shell height, shell thickness and adductor muscle increased after eutrophication began in the late 18th century. After 1860, growth decreased, perhaps reflecting the negative effects of hypoxia, harmful algal blooms, disease and fishing on oyster growth. These results are consistent with the view that an increasing supply of phytoplankton resulting from eutrophication enhanced growth of C. virginica between 1760 and 1860, before oyster reefs were degraded by destructive fishing practices between 1870 and 1930. Alternative factors, such as changes in water temperature, salinity, and fishing are less likely to be responsible for this pattern. These results have implications for restoration of oyster reefs in order to mitigate the effects of eutrophication in estuaries, as well as the paleoecological relationship between suspension feeders and paleoproductivity.     

Occurrence of copepod carcasses in the lower Chesapeake Bay and their decomposition by ambient microbes

We tested and refined the Neutral Red staining method for separating live and dead copepods in natural samples. Live copepods were stained red whereas dead copepods remained unstained. The staining results were not affected by method of killing, time of death or staining time. Tow duration had no significant effect on the percent dead copepods collected. The Neutral Red staining method was applied to study the occurrence of dead copepods along the York River and the Hampton River in the lower Chesapeake Bay during June–July, 2005. The zooplankton community was dominated by copepods; on average 29% of the copepod population appeared dead. Recovery of percent dead copepods did not differ between horizontal tows and vertical tows, suggesting that dead copepods were homogenously distributed in the water column. No significant relationship was found between the percent dead copepods and surface water temperature, salinity, Secchi depth or chlorophyll concentration. In laboratory experiments, dead copepods were decomposed by ambient bacteria and the rate of decomposition was temperature-dependent. Combining field and laboratory results we estimated that the non-consumptive mortality (mortality not due to predation) of copepods in the lower Chesapeake Bay was 0.12 d⁻¹ under steady-state condition, which is within the global average of copepod mortality rate.     

糖原含量与糖原磷酸化酶基因(GPH)和己糖激酶基因(HK)的表达在福建牡蛎生殖周期中的相关性分析

Glycogen, a polymer of glucose, is an important means of storing energy. It is degraded by glycogen phosphorylase (GPH) and hexokinase (HK), glycogen phosphorylase, and hexokinase cDNAs (Ca-GPH andCa-H...     

Biochemical and histological responses in mussel (Mytilus edulis) exposed to North Sea oil and to a mixture of North Sea oil and alkylphenols

Several environmental chemicals are suspected to be responsible for adverse health effects on the reproductive system in various organisms. During this work, environmentally relevant concentrations of North Sea oil were used alone or in combination with alkylphenols and additional PAH to study the effect on vitellogenin-like protein expression and gonadal development in mussels. North Sea oil (0.5 ppm) induced the expression of phospho-proteins in both sexes indicating that some compounds are oestrogen-mimics. This induction was not seen in samples dosed with the mixture but signs of toxic effects were observed in the gonads. Indeed, numerous degenerating ovarian follicles in females and foci, similar to vertebrate melanomacrophage centres, were observed in testes.     

Seasonal variations of alkenones and U37 in the Chesapeake Bay water column

Alkenone unsaturation indices (U^K₃₇ and U^K′₃₇) have long been used as proxies for surface water temperature in the open ocean. Recent studies have suggested that in other marine environments, variables other than temperature may affect both the production of alkenones and the values of the indices. Here, we present the results of a reconnaissance field study in which alkenones were extracted from particulate matter filtered from the water column in Chesapeake Bay during 2000 and 2001. A multivariate analysis shows a strong positive correlation between U^K₃₇ (and U^K′₃₇) values and temperature, and a significant negative correlation between U^K₃₇ (and U^K′₃₇) values and nitrate concentrations. However, temperature and nitrate concentrations also co-vary significantly. The temperature vs. U^K₃₇ relationships (U^K₃₇=0.018 (T)−0.162, R²=0.84, U^K′₃₇=0.013 (T)−0.04, R²=0.80) have lower slopes than the open-ocean equations of Prahl et al. [1988. Further evaluation of long-chain alkenones as indicators of paleoceanographic conditions. Geochimica et Cosmochimica Acta 52, 2303–2310] and Müller et al. [1998. Calibration of the alkenone paleotemperature index U^K′₃₇ based on core-tops from the eastern South Atlantic and the global ocean (60°N–60°S). Geochimica et Cosmochimica Acta 62, 1757–1772], but are similar to the relationships found in controlled studies with elevated nutrient levels and higher nitrate:phosphate (N:P) ratios. This implies that high nutrient levels in Chesapeake Bay have either lowered the U^K₃₇ vs. temperature slope, or nutrient levels are the main controller of the U^K₃₇ index. In addition, particularly high abundances (>5% of total C₃₇ alkenones) of the tetra-unsaturated ketone, C_37:4, were found when water temperatures reached 25 °C or higher, thus posing further questions about the controls on alkenone production as well as the biochemical roles of alkenones.     

Assessing neuroprotective P-glycoprotein activity at the blood-brain barrier in killifish (Fundulus heteroclitus) using behavioural profiles

The neuroprotective role of P-glycoprotein, a multixenobiotic resistance transporter (ABCB1/MDR1), in the blood-brain barrier in fish was examined using behavioural toxicological assays. P-glycoprotein acts as cellular efflux pump to prevent substrates from accumulating in the brain, including environmental contaminants such as ivermectin, a common aquaculture pesticide and mammalian anti-parasitic drug. The behavioural toxicological assays were developed to determine the neuropathological effect of ivermectin in killifish (Fundulus heteroclitus). P-glycoprotein function and thus blood-brain barrier integrity can be compromised by chemosensitizers that inhibit transport activity. Fish treated with ivermectin and the P-glycoprotein inhibitor cyclosporin A were significantly more sensitive and succumbed more rapidly to tilting, lethargy, slowing of pectoral-fin movement and loss of haptic-reactivity compared to fish treated with ivermectin-only. P-glycoprotein inhibition is associated with significantly earlier onset and increased mortality in ivermectin-exposed fish. Our results suggest that P-glycoprotein confers resistance against ivermectin-induced behavioural neuropathology and mortality in fish. This assay provides us with a non-invasive tool to study P-glycoprotein function in the blood-brain barrier and evaluate the behavioural effects of potential environmental neurotoxins.     

湛江湾和平水母属2新种(软水母亚纲,锥螅水母目,和平水母科)

采自广东湛江湾海域软水母亚纲(Leptomedusae Claus,1877)、锥螅水母目(Conica Broch,1916)、和平水母科(Eirenidae Haeckel, 1879)、和平水母属(Eirene Eschscholtz,1829) 2新种,即大腺和平水母新种(E. macrogonia Huang, Sun et Liu,sp. nov.)和湛江和平水母新种(E. zhanjiangensis Huang, Zhang et Zao, sp. nov.)。其鉴别特征如下:(1)大腺和平水母,新种;伞扁于半球形,胶质中等厚;胃柄基部塔状,胃柄较长,伸出伞腔口外;垂管长度短于口唇,4个发达口唇边缘呈齿状皱褶;生殖腺发达波状弯曲,着生于辐管下伞部,从胃柄基部延伸到伞缘外;19～24条缘触手,5～8个缘疣,触手基部呈球状,无排泄乳突;每2条触手间或触手和缘疣间有1个平衡囊,每个平衡囊有1～2个平衡石;4条辐管,1条环管。(2)湛江和平水母,新种;伞半球形,胶质厚;胃柄基部呈塔形,末端变窄;垂管较长,约等于口唇长度,口唇边缘有齿状皱褶;生殖腺带状,从胃柄基部延伸至近伞缘;缘触手多,70～125条,触手基球有排泄乳突;平衡囊数目约为触手数的1/2;4条辐管,1条环管。     

A 15-month study of zooplankton ingestion by farmed mussels (Mytilus edulis) in Bantry Bay, Southwest Ireland

There is a growing body of evidence to suggest that bivalve molluscs routinely ingest zooplankton. To elucidate further these observations, a 15-month study of zooplankton ingestion by farmed mussels was conducted using mussel long-lines in Bantry Bay, Ireland. Stomach content analysis of the mussels showed that there was evidence of zooplankton ingestion throughout the sampling period, but that highest mean numbers of zooplankters were ingested by mussels in the spring and summer months. Various zooplankton species were present in mussel stomachs. Harpacticoid copepods were found more often in stomach contents than calanoid copepods, probably due to their proximity to the bivalves' inhalent siphons. Barnacle cyprids featured in large numbers in stomach contents, but only for a period of 3 months which broadly corresponded with their pelagic phase. Sizes of ingested zooplankton ranged from 126 μm to 6 mm, but more of the smaller zooplankters (e.g. crustacean nauplii) were ingested. When lengths of ingested copepods were compared with those found in plankton net samples, it was found that the net-sampled copepods were significantly larger than those found in mussel stomachs, suggesting that mussels select for smaller categories within the zooplankton available to them. Soft bodied zooplankton was rarely found in mussel stomachs but their absence may be due to rapid digestion or they may have been destroyed in the preservation process. Ingestion of zooplankton by bivalves is discussed in the context of the impacts mussel farms have on resident zooplankton populations.     

Do endocrine disrupting chemicals threaten Mediterranean swordfish? Preliminary results of vitellogenin and Zona radiata proteins in Xiphias gladius

Endocrine Disrupting Chemicals (EDCs) have the potential to alter hormone pathways that regulate reproductive processes in wildlife and fishes. In this research the hypothesis that Mediterranean top predator species (such as large pelagic fish) are potentially at risk due to EDCs is investigated. These marine organisms tend to accumulate high concentrations of EDCs such as polyhalogenated aromatic hydrocarbons (PHAHs). The potential effects of EDCs on a fish species of commercial interest, the top predator Xiphias gladius (swordfish), were investigated using vitellogenin (Vtg) and Zona radiata proteins (Zrp) as diagnostic and prognostic biomarkers. Dramatic induction of typically female proteins (Vtg and Zrp) was detected by ELISA and Western Blot in adult males of the species. These results are the first warning of the potential risk for reproductive function of Mediterranean top predators, and suggest the need for continuous monitoring of this fragile marine environment.     

Appearance of Chelophyes appendiculata and Abylopsis tetragona (Cnidaria, Siphonophora) in the Bay of Villefranche, northwestern Mediterranean

The vertical and temporal distribution of two calycophoran siphonophores, Chelophyes appendiculata (Eschscholtz, 1829) and Abylopsis tetragona (Otto, 1823) in the Bay of Villefranche (northwestern Mediterranean) was investigated by an analysis of three different planktonic time series. A daily series (1993–1995) showed seasonal peaks of the nectophores of C. appendiculata during spring and particularly in late summer, while the abundance of A. tetragona remained similar throughout the year. A weekly series (1994–1995) showed that C. appendiculata (nectophores and eudoxids) became concentrated above the thermal discontinuity, in the most stratified and warm waters, whereas A. tetragona was collected in large numbers below this discontinuity. A 27-year survey (1966–1993) showed long-term fluctuations of these siphonophore populations, which became abundant in the Bay starting from 1980 and especially after 1984, when the water column grew warm and hypersaline, corresponding to a less rainy period. Temporal (seasonal and long-term) and bathymetric (between 10 and 60 m depth) successions of these two siphonophores were noted in this shallow coastal bay.     

分离自中国胶州湾的褐色马格里夫藻(Margalefidinium fulvescens)的形态学和系统进化特征

The dinoflagellate genus Margalefidinium has been split from Cochlodinium as a new genus recently and Margalefidinium fulvescens is one of the five Margalefidinium species. Margalefidinium fulvescens is toxic and has been reported from the coastal waters of USA, Canada, Mexico, China, Japan, Indonesia, Korea, Pakistan and Spain. Here we provide the morphological and phylogenetic characterization for an isolate of it from the Jiaozhou Bay, Qingdao, China. Our results showed that the vegetative cells were subspherical to ellipsoidal, 34–60 μm in length, and 19–41 μm in width. Both single cell forms and colonies in chains of 2, 4, or 8 cells were observed in cultures, but chain forms with 2 or 4 cells were observed more often in the field samples. The cingulum was rather deep, encircling the cell approximately twice, but the sulcus was rather narrow, surrounding the cell about one turn. The nucleus was spherical and located at the central epicone. The chloroplasts were granular, brownish, and scattered peripherally. An orange pigmented body also appeared in the epicone. The apical groove appeared vase-like as previously described. Under epi-fluorescence microscopy, a pumpkin-like structure was clearly observed, in which cells were embedded. Cells were observed to exit from the structure, which led us to a hypothesis that the structure may provide cells a shelter to avoid predation or to respond to other stresses. The phylogenetic analyses based on partial LSU rDNA sequences indicated that M. fulvescens from the Jiaozhou Bay was grouped with M. fulvescens populations from other origins and closely related to the clade of M. polykrikoides.Our morphological observations and phylogenetic analyses together confirmed the presence of M. fulvescens in China and our monitoring has also observed the species dominant in the dinoflagellate community of the Jiaozhou Bay in the early autumn of 2015, which alerted us to continually monitor this bloom-forming species in the region.     

Short-term consequences of a benthic cyanobacterial bloom (Lyngbya majuscula Gomont) for fish and penaeid prawns in Moreton Bay (Queensland, Australia)

This study examined the phenology and ecological consequences of a benthic filamentous cyanobacterial bloom (Lyngbya majuscula) in Deception Bay (Moreton Bay, Queensland, Australia). Bloom initiation occurred in mid December 1999 and expanded to encompass an 8 km² area by April 2000. Small fish and penaeid prawns (<25 cm total length) were quantitatively sampled through periods designated as before, during and after the bloom using a combination of pop-netting within mangroves and beam trawling over adjacent seagrass beds. Data on larger-bodied fish were compiled from daily fishing logs provided by local commercial fishers. Changes in dry mass of bloom material caught in nets and changes in water chemistry were also measured. Mean concentrations of ammonia-N in residual water within mangroves were several orders of magnitude higher in the affected area than in the control and dissolved oxygen was markedly lower in affected areas. Across the study area, mean density, live mass and number of species declined during the bloom, with fish assemblages using mangroves showing greater decline than assemblages using seagrasses. Response at the species level was highly variable; generally, epibenthic species showed a more sustained decline than demersals. Mean monthly fish catch was significantly lower in bloom than non-bloom years. This study has also demonstrated that throughout the bloom, the affected area continued to support a highly diverse and abundant fish and prawn assemblage, and probably maintained its function as an important nursery habitat for many species.     

荣成湾孔鳐群体线粒体DNA cytb部分序列的遗传多样性分析

对2009 年和2011 年采自荣成湾的孔鳐(Raja porosa)群体共54 尾鱼的线粒体DNA 细胞色素b基因(cytb)部分序列进行测定, 以分析其遗传多样性和群体遗传结构。结果显示, 在所获得的782 bp 序列中, 共检测到11 个单倍型、15 个多态位点, 其单倍型多样性指数(H)为0.498±0.081, 核苷酸多样性指数(π)为0.0018±0.0005, 遗传多样性水平较低。其中, 2009 年样品检测到3 种单倍型、4 个单一变异位点, 其H 和π 值分别为0.378±0.181 和0.0010±0.0006; 2011 年样品检测到9 种单倍型、6 个单一变异位点和6 个简约信息位点, 其H 和π 值分别为0.352±0.086 和0.0020±0.0006。分子方差分析(AMOVA)分析显示它们的遗传变异主要集中在同年度个体间(98.22%), 而不同年度个体间的遗传变异远远小于同年度内个体间的变异。Kimura’s 2-parameter 模型分析得到的2009, 2011 年度孔鳐间的遗传距离为0.0013, 遗传分化系数为0.01778, 也表明2009 年和2011 年孔鳐的遗传分化程度低, 没有明显的遗传变异。中性检验表明, 荣成湾孔鳐群体可能发生过种群扩张。