Enhancement of polycyclic aromatic hydrocarbons in estuarine invertebrates by surface runoff at a decommissioned military fuel depot

Accumulation of polycyclic aromatic hydrocarbons (PAHs) was determined in blue mussels (Mytilus spp.) and shore crabs (Hemigrapsus sp.) at a recently closed military fuel depot in central San Francisco Bay, California. In April 1996, during a period of above average precipitation, specimens were collected at the depot, near the depot, and at sites 10 and 20 km south of the depot. Four weeks after the rains ended, blue mussels were again collected at the depot, and at two additional sites in the central Bay region. In April, total PAHs in mussels from the depot were significantly higher only than that in mussels collected 20 km from the depot; however, seven specific, substituted PAHs were higher at the depot than at all other sites. In June, only two of the 38 PAHs common in mussels in April were detected at the depot; these concentrations were comparable to ambient concentrations in mussels at the Bay. It seemed that bioavailability of PAHs at the depot was enhanced by rainfall, probably due to the mobilization of PAHs via groundwater into the Bay. Concentrations in mussels from chronically contaminated sites were about five times higher than mussels collected from the depot. Low PAH concentrations were detected in shore crabs near the depot, and the highest levels were not associated with the depot. Observed PAH concentrations are discussed in relation to upper trophic organisms.     

Preliminary evaluation of macrophage pinocytosis as a technique to monitor fish health

A recently developed spectrophotometric technique was used to examine the effect of environmental pollutants on pinocytosis by fish macrophages. Spot, Leiostomus xanthurus, and hogchoker. Trinectes maculatus, were captured from the York River, a relatively non-polluted ricer, and from the Elizabeth River, known to be highly contaminated with polynuclear aromatic hydrocarbons (PAHs). Kidney cell types were separated by density gradient centrifugation and single cell suspensions of macrophages were used in in vitro studies. The pinocytic activity of kidney macrophages from spot and hogchoker was determined by means of a colorimetric assay of ingested neutral red dye. It was found that the pinocytic activity of macrophages from spot exposed to contaminated waters did not differ from control values. However, there was a two-fold increase in pinocytic activity in pollutant-stressed hogchoker. It is proposed that hogchoker could serve as an indicator species to monitor pollution effects in the environment.     

A description of chloride cell and kidney tubule alterations in the flatfish Solea senegalensis exposed to moderately contaminated sediments from the Sado estuary (Portugal)

The effects of sediment-bound contaminants on kidney and gill chloride cells were surveyed in juvenile Solea senegalensis exposed to fresh sediments collected from three distinct sites of the Sado Estuary (Portugal) in a 28-day laboratorial assay. Sediments were analyzed for metallic contaminants, polycyclic aromatic hydrocarbons and organochlorines as well as for total organic matter, redox potential and fine fraction. The potential for causing adverse biological effects of each surveyed sediment was assessed by comparison of contaminant levels to available guidelines for coastal sediments, namely the Threshold Effects Level (TEL) and the Probable Effects Level (PEL). The Sediment Quality Guideline Quotient indices (SQGQ) were calculated to compare the overall contamination levels of the three stations. A qualitative approach was employed to analyze the histo/cytopathological traits in gill chloride cells and body kidney of fish exposed to each tested sediment for 0, 14 and 28 days. The results showed that sediment contamination can be considered low to moderate and that the least contaminated sediment (from a reference site, with the lowest SQGQ) caused lesser changes in the surveyed organs. However, the most contaminated sediment (by both metallic and organic xenobiotics, with highest SQGQ) was neither responsible for the highest mortality nor for the most pronounced lesions. Exposure to the sediment presenting an intermediate SQGQ, essentially contaminated by organic compounds, caused the highest mortality (48%) and the most severe damage to kidneys, up to full renal necrosis. Chloride cell alterations were similar in fish exposed to the two most contaminated sediments and consisted of a pronounced cellular hypertrophy, likely involving fluid retention and loss of mitochondria. It can be concluded that sediment contamination considered to be low or moderate may be responsible for severe injury to cells and parenchyma involved in the maintenance of osmotic balance, contributing for the high mortality levels observed. The results suggest that sediment-bound organic contaminants such as PAHs (polycyclic aromatic hydrocarbons) and PCBs (polychlorinated biphenyls) may be very toxic to the analyzed organs, especially the kidney, even when present in low-risk concentrations.     

Diel feeding, daily food consumption and the predatory impact of whitemouth croaker (Micropogonias furnieri) in an estuarine environment

Diel feeding pattern and food consumption of whitemouth croaker (Micropogonias furnieri) were examined and related to seasonal variations in an estuarine environment. To determine diel feeding pattern whitemouth croaker were collected from the Patos Lagoon Estuary over 24 h (～3‐h intervals) and their stomach contents were examined. Food consumption of whitemouth croaker was determined from evacuation rates estimated experimentally and in the field. The results suggested that whitemouth croaker in the Patos Lagoon estuary feed all day long. However, whitemouth croaker showed higher feeding intensity and food consumption during daylight hours when the water transparency increased than in others when water transparency was low. Thus, whitemouth croaker seemed to intensify their feeding when they could see their prey. Daily food consumption was between 0.9 and 5.3% of body wet weight (these being the lowest and highest consumption rates during the winter and summer, respectively). The maximum predation impact of whitemouth croaker on their preferential prey, the tanaid Kalliapseudes schubartii, was 27indiv·m^?2·day^?1. The results suggested that the changes in water transparency seem to affect the diel feeding pattern and food consumption of whitemouth croaker. This could have important consequences for fish growth.     

Accumulation and distribution of biogenic and pollutant hydrocarbons,pcbs and ddt in tissues of western mediterranean fishes

Data on hydrocarbons, PCBs and total DDT in muscle, liver and gonads of Mullas barbatus. Trachurus trachurus, Merluccius merluccius, Engraulis enchrasicholus and Micromesistius poutassou collected along the Catalan Coast (NW Mediterrenean) are reported for the first time. The species and the collection sites—off Barcelona to the north (Palamôs) and to the south (Ehro Delta)—were selected for the assessment of the effects of coastal pollutant sources on commercially valuable organisms.Mullus barbatus reflects the influence of local sources, and is therefore a useful bioindicator for organic pollutants. Despite some scattering of the results, concentrations are in general of the same order of magnitude for juvenile and adult specimens, although the pollutants considered exhibit different biological fates. Hydrocarbons are accumulated primarily in liver, whereas organochlorinated compounds are accumulated in gonads. Liver and muscle contents could reflect acute and chronic pollutants inputs respectively. The accumulation in muscle is more important for aromatic and chlorinated hydrocarbons than for the saturated ones, which are probably metabolized in the liver.     

Partitioning of polycyclic aromatic hydrocarbons between water and particles compared to bioaccumulation in mussels: a harbour case

Water and particles from three sites located in Halifax Harbour, near and further away from combined municipal sewage effluents (CMSE) were analysed for parental and alkylated polycyclic aromatic hydrocarbons (par and alkPAH). Bioavailability of PAH was compared for inter-tidal mussels collected at the same sites in April and November. The PAH fingerprint determined over 9 sampling times covering a period of 19 months differed more between phases (water and particles) and seasons, than between sites. In the spring, more alkPAH associated with diesel and gasoline were detected in the soluble phase, along with more bioaccumulation of alkPAH in inter-tidal mussels. A broader number of parPAH were detected in mussels collected in the fall. The mean sum of dissolved alkPAH concentrations was higher in water at the site closer to raw CMSE than at the other two sites and particles of that site. However, lowest bioconcentration factors (BCF) were determined in mussels of this more contaminated site. Similar biota-particle accumulation factors (BPAF) were determined for parPAH in mussels from the three sites, all lower than the BCF of alkPAH. The study indicates that sewage treatment plants will reduce the amount of parPAH especially larger than fluoranthene and pyrene from being deposited in the harbour; that alk naphthalenes and fluorenes present in water will continue to be discharged; that the disturbance of sediments can make particle-bound PAH available to mussels.     

Toxicity of sea-surface microlayer: Effects of hexane extract on baltic herring (Clupea harengus) and atlantic cod (Gadus morhua) embryos

This study was designed to determine whether contaminated sea-surface microlayer was toxic to marine fish embryos in its ntaive form and as a hexane extract. Developing embryos of Atlantic cod (Gadus morhua) and Baltic herring (Clupea harengus) were exposed to hexane extracts of sea-surface microlayer collected from five locations in the North Sea and Baltic Sea. Extracts from two of these locations produced significant embryos mortality as well as severe deformities in live hatched larvae. A control sample of bulk water collected from 20 cm under the surface and extracted in the same way produced no significant mortality or deformities. Significaant changes in timing of hatching were also observed in those samples which produced embryo toxic effects. A comparison of these data with those obtained from code embryos exposed to unextracted microlayer showed a similar biological effect with both unextracted samples and hexane extracts.Chemical analyses revealed the greatest biological effect in samples with petroleum hydrocarbon concentrations between 180 and > 200 μg liter⁻¹. The bulk water control had 1 μg liter⁻¹ while the three samples that showed no biological activity had 3 to 8 μg liter⁻¹ petroleum hydrocarbons. Phthalic acid esters were detected in four samples and chlorinated hydrocarbons in one, but could not be positively correlated with any of the toxic responses. No other chemical contaminants were detected in the five samples.The data presented here show that some sites contain sea-surface microlayer which can be toxic to marine fish embryos: that Baltic herring and Atlantic cod embryos respond similarly to the toxic effects of contaminated microlayer: and that unextracted microlayer and hexane extract of microlayer produce essentially the same toxic effect(s) if only organic contaminants are considered.     

CYP1A- and CYP3A-immunopositive protein levels in digestive gland of the mussel Mytilus galloprovincialis from the Mediterranean Sea

CYP1A-immunopositive protein can be elevated in response to planar PAHs and PCBs in Mytilus sp. digestive gland whilst CYP3A-immunopositive protein has been associated with testosterone 6β-hydroxylation in fish. Levels of CYP1A- and CYP3A-immunopositive protein were determined in Mytilus galloprovincialis digestive gland microsomes collected from 12 sites in the Mediterranean Sea during May and September 2001. CYP1A-immunopositive protein was significantly highest at contaminated sites whilst CYP3A-immunopositive protein was significantly lowest. A weak negative correlation (r²=0.21) was seen between CYP1A- and CYP3A-immunopositive protein. Little evidence of differences at the different sampling times was observed. These results confirm previous work indicating elevation of CYP1A-immunopositive protein in Mytilus sp. digestive gland at contaminated sites. Further study is required to characterise CYP3A-like expression in Mytilus and to elucidate the consequences of possible CYP3A-like down-regulation at contaminated sites.     

Organic and trace metal contaminants in sediments and English sole tissues from Vancouver Harbour,Canada

As part of a multinational workshop on marine environmental quality, sediments were collected from seven sites in Vancouver Harbour and analyzed for polynuclear aromatic hydrocarbons (PAHs) and organochlorines (OCs), and for trace metals. English sole were collected from five sites, and muscle was analyzed for trace metals and liver for OCs. As expected, sediment PAH and OC concentrations and tissue OC concentrations were higher at sites east of the First Narrows, compared to the outer harbor and reference sites. Sediment PAH concentrations east of the First Narrows were similar to concentrations at moderately contaminated sites in Puget Sound, south of Vancouver Harbour. In contrast, concentrations of OCs in sediments and tissue were low to moderate, even at relatively contaminated sites within Vancouver Harbour. Although several trace metals in sediments were higher than in contaminated sediments from Puget Sound, trace metals measured in fish muscle were lower.     

Chlorinated hydrocarbons and hatching success in Baltic herring spring spawners

Eggs from 69 females of spring spawning herring from the German Baltic coast (Travemünde, April 1979) were incubated in clean sea water (20‰ S, temperature 8°C) under standard conditions. Sixty-one trials could be used for the evaluation of hatching success. Viable hatch was taken as a measure to evaluate the effects of chlorinated hydrocarbons accumulated in gonads, liver and muscle of parental fish.PCB levels in running ripe females ranged on a wet weight basis between 19 and 241 ng g^?1 (gonad), 20 and 377 ng g^?1 (liver) and 11 and 1820 ng g^?1 (muscle). Concentrations of other chlorinated hydrocarbons (DDD, DDE, γ-HCH, etc.) were in the same range as reported by other authors for Baltic herring (Huschenbeth, 1973, 1977). Viable hatch was significantly affected at ovary DDE concentrations higher than 18 ng g^?1 (wet wt) and PCB concentrations of more than 120 ng g^?1 (wet wt).Results are compared with data obtained during earlier investigations with flounder eggs.     

Reproductive life history stages sensitive to oil and naphthalene in Atlantic croaker

The reproductive effects of chronic exposure to water-soluble fractions (WSF) of diesel fuel oil and naphthalene in female Atlantic croaker during critical periods of their reproductive life history were investigated. Fish were repeatedly exposed to WSF of oil (2.5 or 5%) or naphthalene (0.5 or 1.0 ppm) for 5 or 8 weeks. Both pollutants blocked sexual maturation in some fish and impaired ovarian recrudescence in others. The majority of the oocytes in exposed fish were undeveloped and widespread oocyte atresia was evident at higher concentrations of these polycyclic aromatic hydrocarbons (PAH). The disruption of ovarian growth and oocyte development in croaker after PAH exposure was associated with alterations in reproductive endocrine function at multiple sites on the hypothalamus-pituitarygonadal axis. A third period of increased sensitivity to xenobiotics was observed during final oocyte maturation.     

Factors affecting the distribution of the bivalve, Macoma balthica (L.) on a mudflat receiving sewage effluent,Fraser river estuary,British Columbia

An investigation of the factors responsible for the distribution of Macoma balthica (L.) on a mudflat receiving sewage effluent was carried out in the Fraser River estuary, British Columbia. The factors examined included reduced salinity, low levels of dissolved oxygen, effluent toxicity, chlorination, substrate grain size and the effects of metal contaminated sediments. Results showed that salinity, dissolved oxygen and sediment grain size did not satisfactorily explain the distribution of M. balthica. Unchlorinated and chlorinated sewage effluent were shown to be non-toxic in laboratory tests, and these results were confirmed by 7-day in situ bioassays. The factor which appeared to determine the distribution of M. balthica was the degree of contamination of the substrate which affected the settling and survival of larval and juvenile clams. Heavy metals occurred in high concentrations in the contaminated sediments and were considered to be the most probable controlling factor.     

Responses of meiofauna and nematode communities to crude oil contamination in a laboratory microcosm experiment

We examined the effects of crude oil contamination on community assemblages of meiofauna and nematodes after exposure to total petroleum hydrocarbons in the laboratory. We administered a seawater solution that had been contaminated with total petroleum hydrocarbons to seven treatment groups at different concentrations, while the control group received uncontaminated filtered seawater. The average density of total meiofauna in the experimental microcosms diluted with 0.5%, 1%, 2%, and 4% contaminated seawater was higher than the density in the control. The average density of total meiofauna in the 8%, 15%, and 20% microcosms was lower than the density in the control. The density of nematodes was similar to that of the total meiofauna. Cluster analysis divided the microcosms into group 1 (control, 0.5%, 1%, 2%, and 4% microcosms) and group 2 (8%, 15%, and 20% microcosms). However, SIMPROF analysis showed no significant difference between the two groups (p > 0.05). Bolbolaimus spp. (37.1%) were dominant among the nematodes. Cluster analysis showed similar results for nematode and meiofaunal communities. The total meiofaunal density, nematode density, and number of Bolbolaimus spp. individuals were significantly negatively associated with the concentration of total petroleum hydrocarbons (Spearman correlation coefficients, p < 0.05). Within the nematodes, epistrate feeders (group 2A: 46%) were the most abundant trophic group. Among the treatment groups, the abundance of group 2A increased in low-concentration microcosms and decreased in high-concentration microcosms. Thus, our findings provide information on the effects of oil pollution on meiofauna in the intertidal zones of sandy beaches.     

黄海小黄鱼生物学特征对多重压力的响应

Temporal changes in biological characteristics of small yellow croaker Larimichthys polyactis in the Yellow Sea were examined for the period of 1960–2008. The body size and age of small yellow croaker decreased substantially, in particular, average length of fish in 2008 was reduced by ~85% than those occurring in 1985, and at that time ~93% of the total catch was dominated by one-year-old individuals. Correspondingly, growth parameters also varied significantly over the years, i.e., k(growth coefficient) and t_0(zero-length age) gradually increased from 0.26 and –0.58 year in 1960 to 0.56 and –0.25 year in 2008, respectively. Although, L∞(body length)sharply decreased from 34.21 cm in 1960 to 24.06 cm in 2008, and t_r(inflexion age) decreased from 3.78 year in1960 to 1.61 year in 2008. There was a great increase both in natural mortality coefficient and fishing mortality coefficient. However, according to the gray correlation analysis, changes in the biological characteristics of small yellow croaker were induced by different stressors ranked as: fishing vessel powerfeeding gradesea surface temperature. This study suggests that the active fishery management measures for biological characters of fish populations should be considered.     

Relationship between hydrocarbons and bacterial activity in mediterranean sediments: Part 2—hydrocarbon degrading activity of bacteria from sediments

In marine sediments from the Mediterranean coast polluted by oil spills (Etang de Berre) high bacterial activity is responsible for self-purification, samples collected from these areas showing a bacterial density several orders of magnitude higher than samples from non-polluted areas (Isle des Embiez). Between 60 and 80 % of the heterotrophic bacteria in the polluted areas are hydrocarbon-degrading, compared with 0.01 % in the non-polluted area.Several strains of hydrocarbon utilisers were isolated, a mixture of strains from each biotope was taken to represent a ‘biotope population’ and their activity towards different types of hydrocarbons (n-alkanes, iso-alkanes, cycloalkanes, aromatics) was determined.With ammonia as the nitrogen source the ‘biotope population’ from the polluted area degraded hydrocarbons much more readily than the ‘biotope population’ from the non-polluted area. With nitrate as the nitrogen source the degradation was much reduced, and, for some hydrocarbons, ceased in both ‘populations’.Individual hydrocarbons encouraged the growth of individual strains in the ‘biotope population’. In natural mixtures of hydrocarbons all strains grew well and degraded aliphatics. There was a good correlation between the respiratory activity of the ‘biotope population’ and the ability of the ‘population’ to utilise hydrocarbons for growth.     

EST-derived microsatellites in Pseudosciaena crocea and their applicability to related species

Simple sequence repeat (SSR) markers were obtained for the large yellow croaker Pseudosciaena crocea using 1 205 expressed sequences tags (ESTs) from the NCBI database. Primers for 48 ESTSSR loci were designed and screened with 30 P.crocea specimens captured from Guanjingyang sea area in Fujian Province of China. Sixteen of the loci were polymorphic, which were amplified with 3 to 11 alleles per locus and the mean of 6.13. The observed and expected heterozygosity per locus ranged from 0.091 to 0.844 (mean 0.544) and from 0.118 to 0.892 (mean 0.644), respectively. Polymorphic information content (PIC) ranged from 0.115 to 0.866 (mean 0.593). The results for cross-species amplification of the 16 large yellow croaker EST-SSRs on P. polyactis, C. niveatus, C. lucidus, A. argentatus and J. belengeri revealed that 14, 12, 11, 7 and 6 loci were successfully amplified with 1 to 10 alleles with an average of 4.5 per locus, respectively, which are suitable for population genetics studies of these species and useful for phylogenetic relationship analysis among these species. Overall, this study provides a set of type I markers for population genetics studies and genome mapping for large yellow croaker and its closely related species.     

Benthic-pelagic coupling in sewage-affected marine ecosystems

Coupling between degraded benthic habitats and populations of epibenthic and pelagic organisms is evident around major sewage outfalls in the Southern California Bight. The distribution of bottom-feeding fishes appears to be affected, but sampling has not been intensive enough to rigorously define patterns for more than a few species. Some evidence suggests that changes in fish abundance may be related to changes in the distribution and abundance of benthic prey; those species (e.g. Dover sole) that can adapt their diets to take advantage of dense populations of opportunistic polychaetes may be found in large numbers around outfalls. Some benthic crustacean feeders are at a disadvantage. Sewage effluent is an allochthnous source of organic matter to the food web and Dover sole and ridgeback shrimp both show shifts in stable isotope ratios consistent with the thesis that sewage-derived organic matter in sediments is trophically transferred to epibenthic organisms. Constructing a carbon budget based on the δ¹³C shifts in these species from area of a major outfall relative to a distant control station suggests that there is an in situ source of isotopically very light carbon in outfall sediments, perhaps from CO₂ fixation by chemoautotrophs and biological methane oxidation. Benthic-pelagic coupling is also evident in the accumulation of chlorinated hydrocarbons by fish and other organisms. At the two major Los Angeles area outfalls log-transformed liver-to-sediment concentration ratios of chlorinated benzenes, which are accumulated to moderate levels in fish, show a linear relationship to the log of K_ow (the octanol-to-water partioning coefficient) of these compounds. Chlorinated hydrocarbons with greater lipid solubility, e.g. PCBs and p,p-DDE, are concentrated to very high levels. Studies of food webs using Cs/K as a measure of trophic level suggest that trophic-step magnification is concentrating these compounds in higher trophic levels.     

Risk of environmental genotoxicity in the Baltic Sea over the period of 2009–2011 assessed by micronuclei frequencies in blood erythrocytes of flounder (Platichthys flesus), herring (Clupea harengus) and eelpout (Zoarces viviparus)

Environmental genotoxicity was investigated at 82 locations encompassing different regions of the Baltic Sea. Micronuclei (MN) analysis was performed in erythrocytes of 1892 specimens of flounder Platichthys flesus, herring Clupea harengus and eelpout Zoarces viviparus, three of the most common native fish species of the Baltic Sea collected in 2009–2011. MN background levels in fish were determined using data obtained in 2001–2011 from 107 Baltic sites. Extremely high genotoxicity risk zones were found for flounder at 11 stations out of 16 in 2009 and 33 stations of 41 in 2010–2011, for herring, at 5 of 18 stations in 2009 and 20 of 43 stations in 2010–2011, in eelpout only at one out of 29 stations. The sampling stations were restricted mainly to the southern and eastern Baltic Sea offshore zones and in most of them, MN frequencies in flounder and herring significantly exceeded the reference and background levels of micronuclei. This is a first attempt to evaluate the background MN responses, as well as low, high and extremely high genotoxicity risk levels for native fish species.     

Nuclear changes in blood, early liver anomalies and hepatocellular cancers in flounder (Platichthys flesus L.) as prognostic indicator for a higher cancer risk?

Nuclear changes (micronuclei) in blood are a well-established biomarker of toxic injury and related genotoxicity in mussel, fish and humans, as well. Yet, the relationship between the frequencies of micronuclei in blood and nuclear changes in liver cells itself and the higher potential risk to develop liver cancer is not yet fully implemented. We analysed and diagnosed macroscopic liver changes and H&E and Feulgen stained histological liver sections (2 microm of thickness) together with blood smears (stained with Sytox Green) from same individuals in flounder caught along a well-known contaminant gradient in the North Sea. Results showed an increase in micronuclei frequencies in blood at the more contaminated sites and higher frequencies of macroscopically visible neoplastic liver changes. In livers of the same individuals, nuclear changes were diagnosed in early to degenerative liver lesion, extra-focal tissue surrounding neoplastic lesion and various stages during cancer progression from early neoplastic foci, benign adenomas and malignant carcinomas. The aim of this approach is to expand diagnostic criteria, well-established in the frame of BEQUALM to nuclear changes which might help diagnosis of malignancies in doubtful cases.     

大黄鱼选育群体与普通养殖群体部分免疫指标的比较

以本课题组选育的“闽优1 号”及未经选育的普通大黄鱼(Larimichthys crocea)养殖群体为研究对象, 比较了两者的白细胞吞噬活性、血清溶菌酶活性、杀菌活力、白蛋白及免疫球蛋白含量、血清及血细胞超氧化物歧化酶(SOD)活性等免疫指标的差异。结果表明: 选育的“闽优1 号”大黄鱼血清溶菌酶和血细胞SOD 活力显著高于对照组(P<0.05), 而血清中的免疫球蛋白含量低于普通养殖群体。在病原菌攻毒后8 d 内, 选育群体累积死亡率显著低于非选育群体, 提示非特异性免疫在大黄鱼抗病免疫的早期阶段可能起着重要作用。