Effects of natural gas condensate - water accommodated fraction on coral larvae

The toxic effects of the water accommodated fraction (WAF) of a natural gas condensate on the larvae of five brooding coral species of the Family Pocilloporidae were examined in short term (96 h) bioassays. Lethality was observed in some larvae of Seriatopora hystrix and Seriatopora guttatus upon exposure to high concentrations of the condensate WAF, while those of Stylophora pistillata, Pocillopora damicornis and Pocillopora verrucosa did not experience mortality. Furthermore, increasing concentrations of WAF produced either delayed or impeded metamorphosis for all species except P. damicornis. Growth of juveniles, exposed to different WAF treatments for 96 h during their larval/early juvenile stages, was measured after 30 d. Marked decrease in subsequent growth rate (polyp count) was observed for S. hystrix, S. guttatus and S. pistillata with increasing WAF concentration. The results showed differential susceptibility of larvae of closely related coral species to oil stress, with the following sensitivity ranking: S. guttatus>S. hystrix>S. pistillata>P. verrucosa>P. damicornis. Oil exposure during the planktonic, larval stage of susceptible corals can adversely affect survivorship, recruitment and/or subsequent colony growth, thereby having profound consequences for the abundance of these corals in space and time.     

Requirement of water-flow for sustainable growth of Pocilloporid corals during high temperature periods

Reef-building corals are threatened worldwide by mass-scale coral bleaching episodes that are pronounced in high sea surface temperature (SST) conditions. Although water-flow has been suggested to be a mitigating factor for bleaching, long-term effects of flow-mediated bleaching suppression are as yet not fully understood. In order to investigate flow effects, we monitored the corals Pocillopora damicornis and Stylophora pistillata grown for 20 months in experimental outdoor flumes with the flow rates of 20 cms(-1) (flow) and <3 cms(-1) (still). Although bleaching was observed under high SST conditions, both species showed a shorter period or entirely no visible bleaching under the flow conditions. Better colony growth was found in the flow conditions whereas significant growth suppression and higher mortality were observed in still conditions. We conclude that water-flow is an essential environmental factor for the corals P. damicornis and S. pistillata, especially under high SST conditions.     

Effects of the herbicide diuron on the early life history stages of coral

The effects of the herbicide diuron on the early life history stages of broadcast spawning and brooding corals were examined in laboratory experiments. Fertilisation of Acropora millepora and Montipora aequituberculata oocytes were not inhibited at diuron concentrations of up to 1000 microg l(-1). Metamorphosis of symbiont-free A. millepora larvae was only significantly inhibited at 300 microg l(-1) diuron. Pocillopora damicornis larvae, which contain symbiotic dinoflagellates, were able to undergo metamorphosis after 24 h exposure to diuron at 1000 microg l(-1). Two-week old P. damicornis recruits on the other hand were as susceptible to diuron as adult colonies, with expulsion of symbiotic dinoflagellates (bleaching) evident at 10 microg l(-1) diuron after 96 h exposure. Reversible metamorphosis was observed at high diuron concentrations, with fully bleached polyps escaping from their skeletons. Pulse amplitude modulation (PAM) chlorophyll fluorescence techniques demonstrated a reduction in photosynthetic efficiency (Delta F/F(m)') in illuminated P. damicornis recruits after a 2 h exposure to 1 microg l(-1) diuron. The dark-adapted quantum yields (F(v)/F(m)) also declined, indicating chronic photoinhibition and damage to photosystem II.     

Effects of whole-stream nitrogen enrichment and litter species mixing on litter decomposition and associated fungi

Nutrient enrichment and changes in riparian tree species composition affect many streams worldwide but their combined effects on decomposers and litter decomposition have been rarely assessed. In this study we assessed the effects of experimental nitrogen (N) enrichment of a small forest stream on the decomposition of three leaf litter species differing in initial chemical composition [alder (Alnus glutinosa), chestnut (Castanea sativa) and poplar (Populus nigra)], incubated individually and in 2-species mixtures during late spring-early summer. To better understand the effects of litter mixing on litter decomposition, component litter species were processed individually for remaining mass and fungal reproductive activity. Litter decomposition rates were high. Nitrogen enrichment significantly stimulated litter decomposition only for alder incubated individually. Differences among litter treatments were found only at the N enriched site where the nutrient rich alder litter decomposed faster than all other litter treatments; only at this site was there a significant relationship between litter decomposition and initial litter N concentration. Decomposition rates of all litter mixtures were lower than those expected from the decomposition rates of the component litter species incubated individually, at the N enriched and reference sites, suggesting antagonistic effects of litter mixing. Conidial production by aquatic hyphomycetes for each sampling date was not affected by nutrient enrichment, litter species or mixing. Aquatic hyphomycetes species richness for each sampling date was higher at the N enriched site than at the reference site and higher for alder litter than for chestnut and poplar, but no effect of mixing was found. Aquatic hyphomycetes communities were structured by litter identity and to a lesser extent by N enrichment, with no effect of mixing. This study suggests that nutrient enrichment and litter quality may not have such strong effects on decomposers and litter decomposition in warmer seasons contrary to what has been reported for autumn-winter. Changes in the composition of the riparian vegetation may have unpredictable effects on litter decomposition independently of streams trophic state.     

Algal diets and natural xenobiotics impact energy allocation in cladocerans. I. Daphnia magna

Recently emerged indications imply that particular concentrations and qualities of dissolved organic carbon (DOC) have the potential to reduce species diversity in zooplankton. In this contribution, we tested the impact of a major component of DOC, the dissolved humic substances (HSs), on two life history traits of a clone of Daphnia magna, namely lifespan and reproduction. We chose reproduction as a variable since it guarantees the persistence of the population in its habitat, and lifespan was selected as the most integrative individual variable. We found that lifespan was extended by exposure to HSs, but lifetime reproduction was clearly reduced. On the other hand, the mineral and biochemical composition of food also significantly modulated these life history traits. By feeding coccal green algae that contained linolenic acid, arginine (Arg), and histidine (His), but that lacked polyunsaturated fatty acids (PUFAs) with C-back bones >18, we examined how short-chained PUFAs, either alone or in concert with Arg and His, are able to modulate life history traits. Our expectation was that the additional challenge by HSs would shift the energy allocation in favor of reproduction. We found that the carbon and amino acid content may account for longevity, even when challenged by HSs. Reproduction appeared to be triggered by the C:P and N:P ratios of the food, particularly given the rather low P-content, which was within the range of severe growth limitation. Consequently, exposure to HSs reduced offspring numbers. Resting eggs without males occurred only in Desmodesmus armatus fed Daphnia that were simultaneously exposed to HSs. Under these conditions, the total amino acid (AA) content seemed to trigger reproduction. In summary, exposure to HSs appeared to have the potential to change the competitive ability of zooplankters by modulating the energy allocation between body maintenance and reproduction.     

The effect of petroleum hydrocarbons on reproduction of an estuarine planktonic copepod in laboratory cultures

The effect of short-term exposure to high concentration (mg l.^?1) of water soluble fraction of aromatic heating oil on subsequent egg production by the estuarine copepod Eurytemora affinis has been studied in laboratory cultures to investigate possibly sublethal biological effects following exposure to hydrocarbon under an oil spill. Significant reduction in subsequent length of life, total number of eggs produced, mean brood size, and rate of egg production was observed. Exposure to naphthalene alone at 1 mg l.^?1 for 24 h produced a significant effect on total fecundity of the females. Ingestion rates were significantly lower when measured in the presence of the water soluble fraction, and also naphthalene alone at mg l.^?1 concentrations. Exposure to low-levels (10 and 50 μg l.^?1) of ¹⁴C-naphthalene alone over a period of ten days produced no significant effect on feeding or reproduction despite the high concentrations of hydrocarbon accumulated during this period. The results are discussed in relation to previous work on the fate of hydrocarbons in planktonic copepods, using ¹⁴C-1-naphthalene as a model compound.     

太湖秀丽白虾生物学与食性的研究

对太湖秀丽白虾怀卵亲体进行培养,两次共获苗1．35万余尾,在实验室内成功地实成了是苗至亲体一个生命周期的饲养,同期结合对太湖白虾的定期采样测定,分析描述了太湖白虾的生产与繁残规律,并对其食性进行了定性与定量的研究分析。     

三峡水库十年生态调度(2011—2020年)期间下游沙市江段产漂流性卵鱼类自然繁殖变化

2011—2020年,三峡水库共开展了14次促进坝下江段四大家鱼自然繁殖的生态调度试验,于每年5—7月在长江中游沙市江段开展鱼类早期资源调查研究,旨在掌握生态调度实施以来产漂流性卵鱼类自然繁殖的响应变化,评估生态调度对长江中游鱼类自然繁殖的贡献。十年间共采集到鱼卵35种,其中典型产漂流性卵鱼类有25种,主要种类包括贝氏?(Hemiculter bleekeri)、鯿(Parabraims pekinensis、银鮈(Squalidus argentatus)、草鱼(Ctenopharyngodon idellus)、鲢(Hypophthalmichthys molitrix)、翘嘴鲌(Culter alburnus)和花斑副沙鳅(Parabotia fasciata);鱼卵群落多样性指数H、均匀度指数E和稳定性指数ICV在年际间呈现先降低后升高的变化趋势,优势度指数D恰好相反,变化拐点出现在2017年,表明2017年以来群落多样性趋于增加,群落结构趋于稳定;产漂流性卵鱼类繁殖规模总体呈先减少后增加变化,有8个类群的繁殖规模逐年增加,鱼类繁殖高峰频次逐渐升高,大规模繁殖出现的时段逐渐延长...     

Effects of elevated CO2 on the reproduction of two calanoid copepods

Some planktonic groups suffer negative effects from ocean acidification (OA), although copepods might be less sensitive. We investigated the effect of predicted CO₂ levels (range 480–750 ppm), on egg production and hatching success of two copepod species, Centropages typicus and Temora longicornis. In these short-term incubations there was no significant effect of high CO₂ on these parameters. Additionally a very high CO₂ treatment, (CO₂ = 9830 ppm), representative of carbon capture and storage scenarios, resulted in a reduction of egg production rate and hatching success of C. typicus, but not T. longicornis. In conclusion, reproduction of C. typicus was more sensitive to acute elevated seawater CO₂ than that of T. longicornis, but neither species was affected by exposure to CO₂ levels predicted for the year 2100. The duration and seasonal timing of exposures to high pCO₂, however, might have a significant effect on the reproduction success of calanoid copepods.     

Hypoxia and benthic community recovery in Korean coastal waters

Low dissolved oxygen (hypoxia and/or anoxia) has become a major cause of change to the benthic component of ecosystems around the world. We present the response of a benthic community to hypoxia in organically enriched environments in Korean coastal waters. Disturbances due to low dissolved oxygen (DO), and organic enrichment altered community dynamics, result in defaunation during summer hypoxia with delayed recolonization occurring in winter. As DO decreased, the number of taxa, their abundance and biomass of macrofauna dropped significantly at inner bay stations in Chinhae Bay and Youngsan River estuarine bay affected by hypoxia. With the return of normoxic conditions in Chinhae Bay, recolonization was initiated by opportunistic species, with a 1-4months lag. The polychaetes, Sigambra tentaculata, Mesochaetopterus sp., and Lumbrineris longifolia, were most persistent under hypoxia. The first recolonizers were the polychaetes Paraprionospio pinnata, S. tantaculata, Glycinde gurjanovae and Nectoneanthes multignatha and the bivalve Theora fragilis. The second group of colonizers included the polychaetes Capitella capitata, Mesochaetopterus sp. and L. longifolia, and the bivalve Raetellops pulchella. Hypoxic and near anoxic conditions resulted in mass mortality in Chinhae Bay and Youngsan River estuarine bay, but communities did partially recover after return to normoxic conditions despite delayed recolonization.     

Spatial distribution of intertidal sandy beach polychaeta along an estuarine and morphodynamic gradient in an eutrophic tropical bay

The spatial distribution of polychaeta along pollution gradients often reflects different degrees of disturbance. In order to evaluate polychaeta fauna of an organically polluted tropical bay, 20 sandy beaches distributed in five areas were sampled. The relationship between community structure, slope, beach index, exposure, sediment and water quality parameters were analysed. Multivariate analysis of variance (PERMANOVA) showed differences among areas and beaches. Scolelepis chilensis dominated at mouth of bay beaches whereas Streblospio gynobranchiata and Capitella capitata complex, at inner beaches. Highest polychaete density was recorded at areas 3 and 5 with the dominance of Saccocirrus sp. and the organic indicator species C. capitata complex and Polydora sp. The most important factors obtained from canonical analysis were sorting, slope, mud and organic matter percentage. Marine biotic index (AMBI) showed that areas 3 and 5 were highly affected by anthropogenic factors, given that a poor polychaeta fauna, dominated by opportunistic species, were found. Polychaete assemblages were affected by eutrophication along an estuarine gradient as well as by morphodynamic condition of the beaches.     

Ammonium,nitrate and phytoplankton interactions in a freshwater tidal estuarine zone: potential effects of cultural eutrophication

Nitrate and ammonium are the most important nitrogen sources for phytoplankton growth. Differential utilization of inorganic nitrogenous compounds by phytoplankton has been observed and may have significant impacts on primary productivity at local scales. We used enrichment experiments with natural phytoplankton populations from the freshwater tidal zone of the Guadiana estuary, a coastal ecosystem increasingly subjected to anthropogenic influences, to study the effects of nitrate and ammonium on N-consumption and phytoplankton growth. In addition, we used combined additions of nitrate and ammonium to understand the inhibitory effect of ammonium over nitrate uptake. Ammonium concentrations in the freshwater tidal reaches of the Guadiana estuary throughout the sampling period were too low to exert an inhibitory effect on nitrate uptake or a toxic effect on phytoplankton growth. Nitrate was clearly the main nitrogen source for phytoplankton at the study site. Overall, nitrate seemed to become limiting at concentrations lower than 20 μM and N-limitation was particularly significant during summer. A trend of decreasing nitrate uptake with increasing ammonium concentrations and uptake suggested an overall preference for ammonium. However, preference for ammonium was group-specific, and it was observed mainly in green algae and cyanobacteria. In fact, cyanobacteria relied only on ammonium as their N-source. On the contrary, diatoms preferred nitrate, and did not respond to ammonium additions. The increasing eutrophication in the Guadiana estuary and particularly increased inputs of nitrogen as ammonium due to urban waste effluents may result in a shift in phytoplankton community composition, towards a dominance of cyanobacteria and green algae.     

A mesocosm experiment investigating the effects of substratum quality and wave exposure on the survival of fish eggs

In a mesocosm experiment, the attachment of bream (Abramis brama) eggs to spawning substrata with and without periphytic biofilm coverage and their subsequent survival with and without low-intensity wave exposure were investigated. Egg attachment was reduced by 73% on spawning substrata with a natural periphytic biofilm, compared to clean substrata. Overall, this initial difference in egg numbers persisted until hatching. The difference in egg numbers was even increased in the wave treatment, while it was reduced in the no-wave control treatment. Exposure to a low-intensity wave regime affected egg development between the two biofilm treatments differently. Waves enhanced egg survival on substrata without a biofilm but reduced the survival of eggs on substrata with biofilm coverage. In the treatment combining biofilm-covered substrata and waves, no attached eggs survived until hatching. In all treatments, more than 75% of the eggs became detached from the spawning substrata during the egg incubation period, and <1% of these detached eggs survived within the substratum interstices. Hence, detached eggs contributed little to the reproductive success we observed. The implications of these results on the spawning success of fish species that use freshly inundated spawning substrata in shallow waters are discussed.     

Integrated use of biomarkers (acetylcholinesterase and antioxidant enzymes activities) in Mytilus galloprovincialis and Mullus barbatus in an Italian coastal marine area

The use of biomarkers to evaluate the biological effects of chemical pollutants in marine organisms represents a recent tool in the monitoring field responding to the need to detect and assess the effects of chemical contaminants on the biota. The aim of the present work was the field application of the integrated use of acetylcholinesterase (AChE) and antioxidant enzymes (catalase--CAT, glutathione peroxidase--GSH-Px), for detecting the possible exposure/effect induced by chemical pollutants in native marine organisms from a coastal marine area, represented by Salento Peninsula (Italy), that shows a coastline of high environmental value, but under constant urban pressure, including agriculture activities, widely diffused in the whole hinterland. Eight sampling stations were chosen: four not urbanized areas considered "uncontaminated" controls and four clearly exposed to anthropogenic impact. The bioindicator species studied were a sessile invertebrate, Mytilus galloprovincialis, and a benthic teleost fish, Mullus barbatus.AChE activity in M. galloprovincialis revealed significant differences among places; the minimum values observed (3.9+/-1.8 nmolmin(-1)mg(-1)) was about 50% reduced with respect to the maximum found (11.4+/-0.9 nmolmin(-1)mg(-1)). The reduction in AChE activity observed in two control stations could be explained by the leaching of pesticides into the sea from the agricultural lands. Moreover, the inhibition of AChE activity by heavy metals besides pesticides, can also explain the reduction of the enzymatic activity observed in an industrialized and harbour area. In M. galloprovincialis AChE activity showed a significant inverse correlation with catalase activity but not with glutathione peroxidase that did not significantly change in animals sampled from the eight stations. Also in M. barbatus AChE activity showed significant differences among places; it was inversely correlated with liver GSH-Px activity, but not with catalase activity, that did not show any significantly variation in animals sampled in the different stations.In conclusion, the integrated use of AChE and antioxidant enzymes (catalase or glutathione peroxidase) in M. galloprovincialis and M. barbatus, two species living in different compartment of marine coastal ecosystem, can find a useful application within the framework of marine coastal environment monitoring programs for detecting the possible exposure/effect induced by chemical pollutants, including pesticides, on living marine organisms.     

Complex environmental gradients predict distributions of river-dependent plants in eastern Australia

We used principal components analysis and multiple logistic regression to investigate the relationships between environmental variables and the distributions of 71 species of river-dependent vascular plants in north-eastern New South Wales, Australia. Our analysis defined seven main environmental factors, summarised (in order of decreasing frequency of statistically significant association with species distributions) as exposure, salinity, stream size, stone scarcity, nutrient enrichment, grazing pressure and rockiness. The main environmental correlates of the presence or absence of macrophyte species in our study were broadly similar to those reported elsewhere, but the relatively low apparent importance of nutrients and grazing was unexpected. We were not able to fully separate the effects of climate-related and non-climatic environmental variables because variables of both types loaded strongly on some principal components, but we suggest that both types of variables should be included in models that aim to forecast potential shifts in plant distributions under projected climatic change. Vascular plants have been neglected in monitoring programs for Australian rivers and their conservation requires a better understanding of patterns and trends in distribution and abundance.     

汉江中下游梯级枢纽联合生态调度对产漂流性卵鱼类自然繁殖的效应研究

为促进汉江中下游鱼类繁殖,2018年6月汉江中下游首次实施梯级联合生态调度试验.该研究在汉江中下游钟祥、沙洋、泽口、仙桃江段设置监测断面,采集鱼类早期资源,并在兴隆枢纽泄洪闸上、下开展鱼类溯流集群情况监测,以分析生态调度对促进汉江中下游鱼类繁殖的效果.监测结果表明汉江中下游鱼卵种类共有26种(属),其中产漂流性卵鱼类22种.监测期间漂流性卵径流量为143411万粒,其中四大家鱼卵径流量为4887万粒,占3.41%,推测汉江中下游有6处成规模的产漂流性卵鱼类产卵场.生态调度期间,坝下鱼类溯流集群随兴隆枢纽调度过程发生变化,钟祥和仙桃江段均出现两次鱼类产卵高峰,鱼卵径流量占总径流量的61.66%,并监测到四大家鱼卵,表明梯级联合生态调度结合区间来水,对汉江下游鱼类,特别是四大家鱼的繁殖具有积极的作用.     

Pyrene induces a reduction in midbrain size and abnormal swimming behavior in early-hatched pufferfish larvae

Spills of heavy oil (HO) have an adverse effect on marine life. We have demonstrated previously that exposure to HO by fertilized eggs of the pufferfish (Takifugu rubripes) induces neural disruption and behavioral abnormality in early-hatched larvae. Here, two kinds of polycyclic aromatic hydrocarbons, pyrene and phenanthrene, were selected to examine their toxic effects on larval behavior of another pufferfish species (T. niphobles). Larvae exposed to pyrene or phenanthrene exhibited no abnormalities in morphology. However, those exposed to pyrene but not phenanthrene swam in an uncoordinated manner, although their swimming distance and speed were normal. The optic tectum, a part of the midbrain, of pyrene-exposed larvae did not grow to full size. Thus, these findings are indicated that pyrene might be a contributor to the behavioral and neuro-developmental toxicity, although there is no indication that it is the only compound participating in the toxicity of the heavy oil mixture.     

Influence of a Xenobiotic Mixture (PCB and TBT) Compared to Single Substances on Swimming Behavior or Reproduction of Daphnia magna

Aroclor 1254, a technical PCB mixture (polychlorinated biphenyls) and TBT (tributyltinchloride) are environmental pollutants that cause a broad spectrum of acute toxic and chronic effects in aquatic animals. In this paper, the sensitivity of Daphnia magna to chronic exposure to mixed xenobiotics was evaluated under laboratory conditions. The results show that xenobiotic mixtures (50 % each of the single compounds) were more toxic than individual xenobiotics alone. By measuring behavioral parameters of animals, it becomes evident that exposure to single xenobiotics significantly affects daphnids: exposure led finally to a rapid decrease in mean swimming activity and also caused changes in preferred swimming depth, with daphnids preferring the upper layers of aquaria. The mixture altered the swimming behavior even more strongly compared to the group stressed by single chemicals. Finally, all daphnids sank to the bottom of the aquaria, still alive, but inactive at the end of the exposure period. In addition, we investigated the reproductive capacity (number of newborn per female and day). PCB did not affect the number of newborn significantly, TBT‐stress led to an evidently decreased number of young daphnids and the xenobiotic mixture decreased reproduction even more. In conclusion, we found significant effects of the single compounds as well as approximately additive (swimming behavior) and synergistic (reproduction) effects of the chemical mixture on daphnids indicating the possibility of dramatic ecological consequences of the occurrence of mixed xenobiotic substances in the aquatic environment.     

The effect of CO2 acidified sea water and reduced salinity on aspects of the embryonic development of the amphipod Echinogammarus marinus (Leach)

We investigated the effect of CO₂ acidified sea water (S = 35, 22 and 10_PSU) on embryonic development of the intertidal amphipod Echinogammarus marinus (Leach). Low pH, but not low salinity (22_PSU), resulted in a more protracted embryonic development in situ although the effect was only evident at low salinity. However reduced salinity, not pH, exerted a strong significant effect, on numbers and calcium content of hatchlings. Females exposed to low salinity (10_PSU) did not carry eggs through to hatching. There was no significant difference in the number of viable hatchlings between females cultured in 22 and 35_PSU but the exoskeleton of the juveniles at 22_PSU contained significantly less calcium. Ocean acidification may affect aspects of E. marinus development but exposure to realistic low salinities appear, in the short term, to be more important in impacting development than exposure to CO₂ acidified sea water at levels predicted for 300 years time.     

ENCORE: the effect of nutrient enrichment on coral reefs. Synthesis of results and conclusions

Coral reef degradation resulting from nutrient enrichment of coastal waters is of increasing global concern. Although effects of nutrients on coral reef organisms have been demonstrated in the laboratory, there is little direct evidence of nutrient effects on coral reef biota in situ. The ENCORE experiment investigated responses of coral reef organisms and processes to controlled additions of dissolved inorganic nitrogen (N) and/or phosphorus (P) on an offshore reef (One Tree Island) at the southern end of the Great Barrier Reef, Australia. A multi-disciplinary team assessed a variety of factors focusing on nutrient dynamics and biotic responses. A controlled and replicated experiment was conducted over two years using twelve small patch reefs ponded at low tide by a coral rim. Treatments included three control reefs (no nutrient addition) and three + N reefs (NH4Cl added), three + P reefs (KH2PO4 added), and three + N + P reefs. Nutrients were added as pulses at each low tide (ca twice per day) by remotely operated units. There were two phases of nutrient additions. During the initial, low-loading phase of the experiment nutrient pulses (mean dose = 11.5 microM NH4+; 2.3 microM PO4(-3)) rapidly declined, reaching near-background levels (mean = 0.9 microM NH4+; 0.5 microM PO4(-3)) within 2-3 h. A variety of biotic processes, assessed over a year during this initial nutrient loading phase, were not significantly affected, with the exception of coral reproduction, which was affected in all nutrient treatments. In Acropora longicyathus and A. aspera, fewer successfully developed embryos were formed, and in A. longicyathus fertilization rates and lipid levels decreased. In the second, high-loading, phase of ENCORE an increased nutrient dosage (mean dose = 36.2 microM NH4+; 5.1 microM PO4(-3)) declining to means of 11.3 microM NH4+ and 2.4 microM PO4(-3) at the end of low tide) was used for a further year, and a variety of significant biotic responses occurred. Encrusting algae incorporated virtually none of the added nutrients. Organisms containing endosymbiotic zooxanthellae (corals and giant clams) assimilated dissolved nutrients rapidly and were responsive to added nutrients. Coral mortality, not detected during the initial low-loading phase, became evident with increased nutrient dosage, particularly in Pocillopora damicornis. Nitrogen additions stunted coral growth, and phosphorus additions had a variable effect. Coral calcification rate and linear extension increased in the presence of added phosphorus but skeletal density was reduced, making corals more susceptible to breakage. Settlement of all coral larvae was reduced in nitrogen treatments, yet settlement of larvae from brooded species was enhanced in phosphorus treatments. Recruitment of stomatopods, benthic crustaceans living in coral rubble, was reduced in nitrogen and nitrogen plus phosphorus treatments. Grazing rates and reproductive effort of various fish species were not affected by the nutrient treatments. Microbial nitrogen transformations in sediments were responsive to nutrient loading with nitrogen fixation significantly increased in phosphorus treatments and denitrification increased in all treatments to which nitrogen had been added. Rates of bioerosion and grazing showed no significant effects of added nutrients. ENCORE has shown that reef organisms and processes investigated in situ were impacted by elevated nutrients. Impacts were dependent on dose level, whether nitrogen and/or phosphorus were elevated and were often species-specific. The impacts were generally sub-lethal and subtle and the treated reefs at the end of the experiment were visually similar to control reefs. Rapid nutrient uptake indicates that nutrient concentrations alone are not adequate to assess nutrient condition of reefs. Sensitive and quantifiable biological indicators need to be developed for coral reef ecosystems. The potential bioindicators identified in ENCORE should be tested in future research on coral reef/nutrient interactions. Synergistic and cumulative effects of elevated nutrients and other environmental parameters, comparative studies of intact vs. disturbed reefs, offshore vs. inshore reefs, or the ability of a nutrient-stressed reef to respond to natural disturbances require elucidation. An expanded understanding of coral reef responses to anthropogenic impacts is necessary, particularly regarding the subtle, sub-lethal effects detected in the ENCORE studies.