Preliminary study on biodegradation of phenanthrene by bacteria isolated from mangrove sediments in Hong Kong

Elevated concentrations of polycyclic aromatic hydrocarbons (PAHs) have been found in mangrove sediments due to anthropogenic pollution, and microbial degradation has been suggested as the best way to remove PAHs from contaminated sediments. The degradation of phenanthrene, a model PAH compound by bacteria, either the enriched mixed culture or individual isolate isolated from surface mangrove sediments was examined. The effects of salinity, initial phenanthrene concentrations and the addition of glucose on biodegradation potential were also investigated. Results show that surface sediments collected from four mangrove swamps in Hong Kong had different degree of PAH contamination and had different indigenous phenanthrene-degrading bacterial consortia. The enriched bacteria could use phenanthrene as the sole carbon source for growth and degrade this PAH compound accordingly. A significant positive relationship was found between bacterial growth and percentages of phenanthrene degradation. The phenanthrene biodegradation ability of the enriched mixed bacterial culture was not related to the degree of PAH contamination in surface sediments. The growth and biodegradation percentages of the enriched mixed culture were not higher than that of the individual isolate especially at low salinity (0 and 10 ppt). High salinity (35 ppt) inhibited growth and biodegradation of phenanthrene of a bacterial isolate but less inhibitory effect was found on the mixed culture. The inhibitory effects of salinity could be reduced with the addition of glucose.     

Dredging related metal bioaccumulation in oysters

Bivalves are regularly used as biomonitors of contaminants in coastal and estuarine waters. We used oysters to assess short term changes in metal availability caused by the resuspension of contaminated sediments. Sydney Rock Oysters, Saccostrea glomerata, were deployed at multiple sites in Port Kembla Harbour and two reference estuaries for 11 weeks before dredging and for two equivalent periods during dredging. Saccostrea experienced large increases in accumulation of zinc, copper and tin during dredging in the Port relative to oysters deployed in reference estuaries. Lead and tin were found to be permanently elevated within Port Kembla. We present a clear and un-confounded demonstration of the potential for dredging activities to cause large scale increases in water column contamination. Our results also demonstrate the usefulness of external reference locations in overcoming temporal confounding in bioaccumulation studies.     

PAHs contamination and bacterial communities in mangrove surface sediments of the Jiulong River Estuary, China

Sixteen sediment samples collected from eight transects in a mangrove swamp of the Jiulong River Estuary, Fujian, China were investigated for their content of polycyclic aromatic hydrocarbons (PAHs) and the biodegradation potential of the indigenous microorganisms. The bacterial community structures in the mangrove sediments and in enrichment cultures were also investigated. The results showed that the total PAHs concentration of mangrove sediments ranged from 280 to 1074 ng g(-1) dry weight, that the PAHs composition pattern in the mangrove sediments was dominated by high molecular weight PAH components (4-6 rings), and that Benzo[ghi]perylene and Indeno[1,2,3-cd]pyrene were the most dominant at different stations. Abundant PAH-degrading bacteria were found in all the stations, the values of phenanthrene-degrading bacteria ranged from 5.85 x 10(4) to 7.80 x 10(5) CFU g(-1) dry weight, fluoranthene-degrading bacteria ranged from 5.25 x 10(4) to 5.79 x 10(5) CFU g(-1) dry weight, pyrene-degrading bacteria ranged from 3.10 x 10(4) to 6.97 x 10(5) CFU g(-1) dry weight and the benzo(a)pyrene-degrading bacteria ranged from 5.25 x 10(4) to 7.26 x 10(5) CFU g(-1) dry weight. DGGE analysis of PCR-amplified 16S rDNA gene fragments confirmed that there was a remarkable shift in the composition of the bacterial community due to the addition of the different model PAH compound phenanthrene (three ring PAH), fluoranthene(four ring PAH), pyrene(four ring PAH) and benzo(a)pyrene(five ring PAH) during enrichment batch culture. Eleven strains were obtained with different morphology and different degradation ability. The presence of common bands for microbial species in the cultures and in the native mangrove sediment DNA indicated that these strains could be potential in situ PAH-degraders.     

Fate of polycyclic aromatic hydrocarbon (PAH) contamination in a mangrove swamp in Hong Kong following an oil spill

The fate of polycyclic aromatic hydrocarbon (PAH) contamination in a mangrove swamp (Yi O) in Hong Kong after an oil spill accident was investigated. The concentrations and profiles of PAHs in surface sediments collected from five quadrats (each of 10 m×10 m) covering different degrees of oil contamination and the most contaminated mangrove leaves were examined in December 2000 (30 days after the accident) and March 2001 (126 days later). The concentrations of total PAHs in surface sediments ranged from 138 to 2135 ng g⁻¹, and PAHs concentrations decreased with time. In the most contaminated sediments, total PAHs dropped from 2135 (30 days) to 1196 ng g⁻¹ (120 days), and the decrease was smaller in less contaminated sediments. The percentage reduction in sediment PAHs over three months (44%) was less significant than that in contaminated leaves (85%), indicating PAH in or on leaves disappeared more rapidly. The PAH profiles were very similar in sediments collected from quadrats Q1 and Q2 with benzo[a]anthracene and pyrene being the most abundant PAH compounds, but were different in the other three quadrats. The proportion of the light molecular weight PAHs to total PAHs increased after three months, especially phenanthrene. Results suggest that physical and photo-chemical weathering (tidal washing and photo-oxidation) of crude oil in surface sediments and on plant leaves were important processes in the first few months after the oil spill. The PAH contamination in Yi O swamp came from both petrogenic and pyrolytic sources. The petrogenic characteristic in the most contaminated sediment was confirmed with high values of phenanthrene to anthracene ratio (>10) and low values of fluoranthene to pyrene ratio (0.3–0.4).     

Modified sublimation to isolate phenanthrene-degrading bacteria of the genera Sphingomonas and Burkholderia from Xiamen oil port

Sublimation was developed by Alley and Brown (2000) in order to isolate bacterial strains that were capable of degrading water insoluble compounds. In this study, sublimation was modified by the use of nutritional agar plates, instead of mineral salt agar, to isolate phenanthrene-degrading bacteria from a mixed culture that had been enriched under the selective pressure of high phenanthrene content. Five strains were obtained with different morphology and degradation ability. Based on the 16S rDNA sequence, two of them were classified as species of the genus Sphingomonas; the others as species of the genus Burkholderia. Denaturing gradient gel electrophoresis (DGGE) was introduced to detect dynamic changes in the bacterial community during enrichment batch culture, and to determine any correlation between the five isolates and the phenanthrene-degrading consortium. The DGGE profile indicated that these five isolates corresponded to four dominant bands of the consortium. Compared to traditional means of isolation, we concluded that modified sublimation is effective and more convenient.     

Prevalence and genetic profiles of Escherichia coli from mangroves and mangrove associated foods off Goa,India

A total of 120 samples comprising of water (45), sediment (45) and mangrove originated food (30) collected from mangrove ecosystems of Goa were screened for Escherichia coli employing ISO-16654 method. Seventy-one (59.16%) samples were positive for E. coli. The E. coli isolates were further characterized by serotyping, virulence gene profiling and pulsed field gel electrophoresis (PFGE). Water and sediment samples were analyzed for physico-chemical parameters. The serotypes reported were O1, O10, O13, O17, O36, O41, O50, O68, O105, O116, O141, O148, O159, O162 and rough types while, 23 strains could not be typed. The stx1 and stx2 genes were detected in 33(46.47%) and 16(22.53%) isolates, respectively. The XbaI restriction digestion patterns of the stx positive strains were diverse. Interestingly, few strains isolated from diarrheal patients and from water, sediment and food from mangrove sources were genetically similar. The study showed that the mangrove ecosystem could be a potential reservoir for pathogenic E. coli.     

Multi-factors on biodegradation kinetics of polycyclic aromatic hydrocarbons (PAHs) by Sphingomonas sp. a bacterial strain isolated from mangrove sediment

Biodegradation of polycyclic aromatic hydrocarbons (PAHs) in contaminated sediment is an attractive remediation technique and its success depends on biodegradation kinetics, and the optimal condition for the PAH-degrading isolates; however, information on this aspect is still scarce. The effects of multi-factors on biodegradation of phenanthrene, a 3-ring model PAH, in contaminated sediment slurry by Sphingomonas sp. a bacterial strain isolated from surface mangrove sediment, were investigated using the orthogonal experimental design (form L₁₆(4⁵)). The most significant factors were salinity and inoculum size, while the effects of phenanthrene concentrations, nutrient addition and temperatures were insignificant. The optimal biodegradation condition in contaminated mangrove sediment slurry was 30 °C, 15 ppt salinity, a carbon/nitrogen ratio of 100:1 (the background ratio in sediment) and an inoculum size of 10⁶ most probable number g⁻¹ sediment. The phenanthrene biodegradation could be best described by the first order rate model, C = C₀e^−kt, where k (the rate constant) is equaled to 0.1185, under the optimal condition. The kinetic model was verified and its validity in predicting biodegradation by Sphingomonas sp. at various phenanthrene concentrations was proved by experimental data.     

Impact of discards of beam trawl fishing on the nematode community from the Tagus estuary (Portugal)

The impact of dead discards, originating from beam trawl fishing on the nematode community from the Tagus estuary was investigated in terms of vertical distribution of the dominant nematode groups. Sediment cores were collected from a mud-flat from the Tagus estuary. Crangoncrangon (Linnaeus, 1758) carcasses were added to the surface of the cores, simulating the settling of dead discards on the sediment. The vertical distribution of the dominant nematode groups was determined up to 4 cm deep at four different moments in time post deposition (0, 2, 4 and 6 h) and compared to control cores. The C.crangon addition to the sediment led to the formation of black spots and therefore oxygen depleted areas at the sediment surface. The Chromadora/Ptycholaimellus group, normally dominant at the surface layer, migrated downwards due to their high sensibility to toxic conditions. Sabatieria presented the opposite trend and became the dominant group at the surface layer. Since Sabatieria is tolerant to oxygen stressed conditions and high sulphide concentrations, we suggest that it migrated opportunistically towards an unoccupied niche. Daptonema, Metachromadora and Terschellingia did not show any vertical migration, reflecting their tolerance to anoxic and high sulphidic conditions. Our study showed that an accumulation of dead discards at the sediment surface might therefore alter the nematode community vertical distribution. This effect is apparently closely related to toxic conditions in the sediment, induced by the deposition of C.crangon at the sediment surface. These alterations might be temporal and reflect an adaptation of the nematode community to dynamic intertidal environments.     

PAHs contamination and bacterial communities in mangrove surface sediments of the Jiulong River Estuary,China

Sixteen sediment samples collected from eight transects in a mangrove swamp of the Jiulong River Estuary, Fujian, China were investigated for their content of polycyclic aromatic hydrocarbons (PAHs) and the biodegradation potential of the indigenous microorganisms. The bacterial community structures in the mangrove sediments and in enrichment cultures were also investigated. The results showed that the total PAHs concentration of mangrove sediments ranged from 280 to 1074 ng g⁻¹ dry weight, that the PAHs composition pattern in the mangrove sediments was dominated by high molecular weight PAH components (4–6 rings), and that Benzo[ghi]perylene and Indeno[1,2,3-cd]pyrene were the most dominant at different stations. Abundant PAH-degrading bacteria were found in all the stations, the values of phenanthrene-degrading bacteria ranged from 5.85 × 10⁴ to 7.80 × 10⁵ CFU g⁻¹ dry weight, fluoranthene-degrading bacteria ranged from 5.25 × 10⁴ to 5.79 × 10⁵ CFU g⁻¹ dry weight, pyrene-degrading bacteria ranged from 3.10 × 10⁴ to 6.97 × 10⁵ CFU g⁻¹ dry weight and the benzo(a)pyrene-degrading bacteria ranged from 5.25 × 10⁴ to 7.26 × 10⁵ CFU g⁻¹ dry weight. DGGE analysis of PCR-amplified 16S rDNA gene fragments confirmed that there was a remarkable shift in the composition of the bacterial community due to the addition of the different model PAH compound phenanthrene (three ring PAH), fluoranthene(four ring PAH), pyrene(four ring PAH) and benzo(a)pyrene(five ring PAH) during enrichment batch culture. Eleven strains were obtained with different morphology and different degradation ability. The presence of common bands for microbial species in the cultures and in the native mangrove sediment DNA indicated that these strains could be potential in situ PAH-degraders.     

Contamination and potential biodegradation of polycyclic aromatic hydrocarbons in mangrove sediments of Xiamen, China

Five stations were established in the Fenglin mangrove area of Xiamen, China to determine the concentrations of polycyclic aromatic hydrocarbons (PAHs) and the numbers of PAH-degrading bacteria in surface sediments. Assessing the biodegradation potential of indigenous microorganisms and isolating the high molecule weight (HMW)-PAH degrading bacteria was also one of the aims of this work. The results showed that the total PAH concentration of sediments was 222.59 ng g(-1) dry weight, whereas the HMW-PAH benzo(a)pyrene (BaP) had the highest concentration among 16 individual PAH compounds. The variation in the numbers of PAH-degrading bacteria was 2.62 x 10(2)-5.67 x 10(4)CFU g(-1) dry weight. The addition of PAHs showed a great influence in increasing the microbial activity in mangrove sediments. A bacterial consortium, which could utilize BaP as the sole source of carbon and energy, and which was isolated from mangrove sediments and enriched in liquid medium for nearly one year degraded 32.8% of BaP after 63 days incubation.     

Degradation of crude oil in a contaminated tidal flat area and the resilience of bacterial community

Crude oil spills, Hebei Spirit in South Korea, is considered as one of the worst environmental disasters of the region. Our understanding on activation of oil-degrading bacteria and resilience of microbial community in oil contaminated sites are limited due to scarcity of such event. In the present study, tidal flat sediment contaminated by the oil spill were investigated for duration of 13 months to identify temporal change in microbial community and functional genes responsible for PAH-degradation. The results showed predominance of previously known oil-degrading genera, such as Cycloclasticus, Alcanivorax, and Thalassolituus, displaying significant increase within first four months of the accident. The disturbance caused by the oil spill altered the microbial community and its functional structures, but they were almost restored to the original state after 13 months. Present study demonstrated high detoxification capacity of indigenous bacterial populations in the tidal flat sediments and its resilience of microbial community.     

Abilities and genes for PAH biodegradation of bacteria isolated from mangrove sediments from the central of Thailand

PAH-degrading bacteria, including Novosphingobium sp. PCY, Microbacterium sp. BPW, Ralstonia sp. BPH, Alcaligenes sp. SSK1B, and Achromobacter sp. SSK4, were isolated from mangrove sediments. These isolates degraded 50–76% of 100 mg/l phenanthrene within 2 weeks. Strains PCY and BPW also degraded pyrene at 98% and 71%, respectively. Furthermore, all of them probably produced biosurfactants in the presence of hydrocarbons. Interestingly, PCY has a versatility to degrade various PAHs. Molecular techniques and plasmid curing remarkably revealed the presence of the alpha subunit of pyrene dioxygenase gene (nidA), involving in its pyrene/phenanthrene degrading ability, located on megaplasmid of PCY which has never before been reported in sphingomonads. Moreover, genes encoding ferredoxin, reductase, extradiol dioxygenase (bphA3A4C) and exopolysaccharide biosynthetase, which may be involved in PAH degradation and biosurfactant production, were also found in PCY. Therefore, we conclude that these isolates, especially PCY, can be the candidates for use as inoculums in the bioremediation.     

Effectiveness of bacterial inoculum and mangrove plants on remediation of sediment contaminated with polycyclic aromatic hydrocarbons

The remediation of mangrove sediment contaminated with mixed polycyclic aromatic hydrocarbons (PAHs) having 3-, 4- and 5-rings by natural attenuation, bioaugmentation, phytoremediation and its combination was compared by greenhouse microcosm studies. At Days 90 and 154, the decreases of PAHs in contaminated mangrove sediment by phytoremediation, planted with one-year old Aegiceras corniculatum, and bioaugmentation, the inoculation of PAH-degrading bacterial strains isolated from mangrove sediment, either SCSH (Mycobacterium parafortuitum) or SAFY (Sphingobium yanoikuyae), were not better than that by natural attenuation (the non-vegetated and un-inoculated microcosms). The populations of SCSH and SAFY in sediment could not be maintained even with repeated inoculation, suggesting that the two isolates were not able to compete with the indigenous microbes and had little enhancement effect. Although some PAHs were accumulated in roots, root uptake only accounted for <15% of the spiked PAHs and the effect of plants on remediation were also insignificant. At the end of the 154-day experiment, the mass balance calculation revealed that the overall losses of PAHs by phytoremediation were comparable to that by bioaugmentation but were lower than that by natural attenuation, especially for the high molecular weight PAHs. Under natural attenuation, around 90% fluorene, 80% phenanthrene, 70% fluoranthene, 68% pyrene and 32% benzo[a]pyrene in contaminated sediment were removed. These results demonstrated that the mangrove sediment itself had sufficient indigenous microorganisms capable of naturally remedying PAH contamination.     

Long-term differences in annual litter production between alien (Sonneratia apetala) and native (Kandelia obovata) mangrove species in Futian,Shenzhen, China

Annual litter production in alien (Sonneratia apetala) and native (Kandelia obovata) mangrove forests in Shenzhen, China were compared from 1999 to 2010. S. apetala had significantly higher litter production than K. obovata, with mean annual total litter of 18.1 t ha⁻¹ yr⁻¹ and 15.2 t ha⁻¹ yr⁻¹, respectively. The higher litter production in S. apetala forest indicates higher productivity and consequently more nutrient supply to the estuarine ecosystems but may be more invasive due to positive plant-soil feedbacks and nutrient availability to this alien species. Two peaks were recorded in S. apetala (May and October), while only one peak was observed in K. obovata, in early spring (March and April). Leaf and reproductive materials were the main contributors to litter production (>80%) in both forests. These results suggest that the ecological function of S. apetala and its invasive potential can be better understood based on a long-term litter fall analysis.     

Effects of copper and the sea lice treatment Slice on nutrient release from marine sediments

Copper-based antifoulant paints and the sea lice treatment Slice^® are widely used, and often detectable in the sediments beneath farms where they are administered. Ten-day, whole sediment mesocosm experiments were conducted to examine how increasing sediment concentrations of copper or Slice^® influenced final water column concentrations of ammonium-nitrogen (NH₄-N), nitrate + nitrite-nitrogen (NO_X-N) and phosphate-phosphorus (PO₄-P) in the presence of the non-target, benthic organisms Corophium volutator and Hediste diversicolor. Nominal sediment concentrations of copper and Slice^® had significant effects on the resulting concentrations of almost all nutrients examined. The overall trends in nutrient concentrations at the end of the 10-day incubations were highly similar between the trials with either copper or Slice^®, irrespective of the invertebrate species present. This suggests that nutrient exchange from the experimental sediments was primarily influenced by the direct effect of copper/Slice^® dose on the sediment microbial community, rather than the indirect effect of reduced bioturbation/irrigation due to increased macrofaunal mortality.     

Impact of the Phaeocystis globosa spring bloom on the intertidal benthic compartment in the eastern English Channel: a synthesis

From 1999 to 2005, studies carried out in the frame of regional and national French programs aimed to determine whether the Phaeocystisglobosa bloom affected the intertidal benthic communities of the French coast of the eastern English Channel in terms of composition and/or functioning. Study sites were chosen to cover most of the typical shore types encountered on this coast (a rocky shore, an exposed sandy beach and a small estuary). Both the presence of active Phaeocystis cells and their degradation product (foam) did have a significant impact on the studied shores. The primary production and growth rates of the kelp Saccharina latissima decreased during the bloom because of a shortage of light and nutrient for the macroalgae. On sandy sediments, the benthic metabolism (community respiration and community primary production), as well as the nitrification rate, were enhanced during foam deposits, in relation with the presence of bacteria and active pelagic cells within the decaying colonies. In estuarine sediments, the most impressive impact was the formation of a crust at the sediment surface due to drying foam. This led to anoxic conditions in the surface sediment and resulted in a high mortality among the benthic community. Some organisms also tended to migrate upward and were then directly accessible to the higher trophic level represented by birds. Phaeocystis then created a shortcut in the estuarine trophic network. Most of these modifications lasted shortly and all the systems considered came back to their regular properties and activities a few weeks after the end of the bloom, except for the most impacted estuarine area.     

Diatom community dynamics in a tropical,monsoon-influenced environment: West coast of India

Diatom communities are influenced by environmental perturbations, such as the monsoon system that impact the niche opportunities of species. To discern the influence of the monsoon system on diatom community structure, we sampled during two consecutive post-monsoons (2001 and 2002) and the intervening pre-monsoon at Mumbai and Jawaharlal Nehru ports along the central west coast of India. Characteristic temporal shifts in diatom community structure were observed across the sampling periods; these were mainly driven by temperature, salinity and dissolved oxygen saturation. The nutrient-poor pre-monsoon period supported low abundance yet high species richness and diversity of diatoms. Coscinodiscus, Cyclotella, Thalassiosira, Triceratium, Pleurosigma, Skeletonema and Surirella were the most dominant genera. Both the post-monsoon periods, following dissimilar monsoon events, were dominated by Skeletonema costatum, but differed in some of the residual species. Thalassiosira and Thalassionema spp. dominated mostly during post-monsoon I whereas Triceratium and Pleurosigma spp. dominated during post-monsoon II. To understand the underlying ecological mechanisms involved in such dynamics, we focus on the dominant diatom species in post-monsoon periods, S. costatum, that contributes up to 60% to total diatom cell numbers. This research is relevant in light of the fluctuating monsoon regimes over the Asian continent, the confounding effects of anthropogenic eutrophication and the resulting cascading effects on trophic web dynamics.     

Chlorella-bearing ciliates (Stentor, Ophrydium) dominate in an oligotrophic, deep North Patagonian lake (Lake Caburgua, Chile)

The aim of this study was to present quantitative data on the population dynamics of Chlorella-bearing ciliates (Stentor, Ophrydium) compared to the total zooplankton community in a deep, oligotrophic North Patagonian lake. Mixotrophic and heterotrophic ciliates, rotifers and microcrustaceans, and important ecological parameters were sampled during a 1-year study. The results showed a low biodiversity with only a few dominant species in every zooplankton group. Three mixotrophic ciliates - Stentor araucanus, S. amethystinus and Ophrydium naumanni - were found. They peaked in summer and autumn with maximum values of 152-313 Ind L⁻¹ (Stentor) and 1880 Ind L⁻¹ (Ophrydium). Their contribution to the total ciliate abundance was 16±17% (annual average). Both Stentor species displayed a distinct vertical zonation during the stratification period with peak depth between 10 and 15 m (metalimnion). The contribution to total zooplankton biomass was 59.4% on an annual average (Stentor: 41%, O. naumanni: 18.4%) and 83% during the stratification period. Both abundance and biomass of mixotrophic ciliates correlated strongly with temperature and to a lesser degree with copepods, rotifers and small cladocerans. According to this study mixotrophic ciliates were by far the dominant zooplankton group in Lake Caburgua. We report for the first time the importance of O. naumanni in a deep Chilean North Patagonian lake.     

Biomarker responses in pelagic and benthic fish over 1 year following the Hebei Spirit oil spill (Taean, Korea)

After the Hebei Spirit oil spill incident (7th December, 2007) in the west coast of Korea, contamination of biliary PAH metabolite and hepatic biomarkers in a pelagic and a benthic fish was monitored for 1 year. Concentrations of 16 PAHs and alkylated PAHs in fish muscle were highest (22.0 ng/g d.w. for 16 PAHs and 284 ng/g d.w. for alkylated PAHs) at 5 days after the spill and then decreased rapidly to background levels at 11 months after the spill. Fish from the oiled site had elevated biliary PAH metabolite concentrations immediately after the spill; these declined steadily in both species, but were still above reference site concentrations 2 months after the spill. Oiled-site fish showed hepatic CYP 1A induction whose trend closely followed those of biliary PAH metabolite concentrations, implying continuous exposure to PAHs. Brain acetylcholinesterase activity was not related to oil exposure.     

Application of bioassays in toxicological hazard, risk and impact assessments of dredged sediments

Given the potential environmental consequences of dumped dredged harbour sediments it is vital to establish the potential risks from exposure before disposal at sea. Currently, European legislation for disposal of contaminated sediments at sea is based on chemical analysis of a limited number of well-known contaminants for which maximum acceptable concentrations, action levels (ALs), have been set. The present paper addresses the issue of the applicability of in vitro and in vivo bioassays for hazard, risk and local impact assessment of dredged polluted sediments to be disposed of at sea. It discusses how and to what extent selected bioassays can fill in the gaps left open by chemical analysis and the way in which the bioassays may contribute to the present licensing system for disposal. Three different purposes for application were distinguished: the most basic application (A) is a rapid determination of the hazard (potential toxicity) of dredged sediments which is then compared to ALs in a licensing system. As with chemical analysis on whole sediment extracts, the bioavailability of the chemicals is not taken into account. As in vitro assays with sediment extracts are not sensitive to matrix effects, a selection of specific in vitro bioassays can be suitable fast and standardized additions for the licensing system. When the outcome of (A) does not convincingly demonstrate whether the sediment is clean enough or too polluted, further bioanalysis can help the decision making process (B). More aspects of the mostly unknown complex chemical mixtures are taken into account, including the bioavailability and chronic toxicity focusing on ecologically relevant endpoints. The ecotoxicological pressure imposed by the dredged sediments can be quantified as the potentially affected fraction (PAF) based on chemical or biological analysis of levels of contaminants in sediment or biota. To validate the predicted risk, the actual impact of dumped harbour sediments on local ecosystems (C) can be determined using a dedicated set of in vitro and in vivo bioassays as well as bio-indicators selected based on the information obtained from (A) and (B) and on the characteristics of the local ecosystem. Conversely, the local sediment impact assessment (C) can direct fine-tuning of the selection of chemical and bioassay analyses and for setting safe levels in the licensing system. It is concluded that in vitro and in vivo bioassays and biological indicators are useful tools in the process of hazard, ecotoxicological risk and impact assessment of dredged harbour sediments, provided they are consciously chosen and quality criteria for assay performance are defined.