The viviparous blenny (Zoarces viviparus) as a bioindicator of contaminant exposure: application of biomarkers of apoptosis and DNA damage

The viviparous blenny (Zoarces viviparus), also known as eelpout, is considered a suitable biomonitor for use in European estuarine and coastal waters. We have previously shown that Z. viviparus from the industrialised Tyne estuary (UK) display certain histopathologies (e.g., ovotestis and hepatic nuclear and cellular pleomorphism), which are associated with contaminant exposure [Mar. Environ. Res. 55 (2003) 137]. Furthermore, the prevalence of these pathologies was higher than in fish collected from a less contaminated reference site (the Alde estuary, UK). Here, tissue samples were collected from Z. viviparus from the Tyne and Alde estuaries (in the spring and autumn) and analysed for DNA adducts using the (32)P-postlabelling assay and for histopathology. Z. viviparus caught from the Tyne during the spring (56.4+/-18.5 adducts per 10(8) nucleotides) and autumn (63.1+/-7.5 adducts per 10(8) nucleotides) exhibited patterns of DNA adducts which indicated exposure to a complex mixture of genotoxins. In contrast, levels of DNA adducts in Z. viviparus from Alde estuary were significantly lower during both sampling periods. Histopathological changes to the gonads included intersex (ovotestis) and germ cell apoptosis (detected via the TUNEL assay) in male fish captured from the Tyne estuary. Further studies on the use of germ cell apoptotic rate at different estuarine sites are required if it is to be used as a reproductive biomarker of contaminant exposure in male fish.     

Pathology of the liver, kidney and gonad of flounder (Platichthys flesus) from a UK estuary impacted by endocrine disrupting chemicals.

The Mersey estuary is highly contaminated with xenobiotics compared to the nearby Dee estuary. Male flounder, a migratory flatfish caught in the Mersey frequently contains high blood concentrations of the oestrogen controlled, female protein vitellogenin, suggesting that Mersey flounder have been exposed to endocrine disrupting contaminants. Males caught from the Dee contain lower blood vitellogenin levels. Preliminary histopathological examination of 410 flounder from both estuaries, focuses on the liver, kidney and gonads. Hepatic tubular vacuolation, foci of cellular alteration, and hepatocellular tumours were seen. Renal and gonadal pathology included the presence of two Mersey fish with enlarged and abnormal glomeruli, three phenotypic male Mersey flounder with unilateral intersex gonads and one male with bilateral, abnormal ovarian follicular components in the testis. These initial results provide pathological evidence of xenobiotic exposure in flounder sampled from both estuaries. Preliminary findings from flounder sampled from Millport, an offshore site in Southwest Scotland, showed no histopathological evidence of xenobiotic exposure.     

Differential sensitivity of flounder (Platichthys flesus) in response to oestrogenic chemical exposure: an issue for design and interpretation of monitoring and research programmes

This study was conducted as an initial investigation of 'differential response' in one of the main sentinel organisms used for monitoring programmes in United Kingdom estuaries, the flounder Platichthys flesus. It has been hypothesised that monitoring using species with a wide geographical spread and limited migration, such as flounder, might result in the comparison of different genetic stocks and certainly of populations with differing early life stage contaminant exposure histories. Furthermore, it is probable that these pre-exposure and genetic differences could manifest themselves in an ability to respond differently to contaminant exposure, so-called 'differential response'. It is important that the extent and nature of this response is understood, if we want to be able to fully interpret the monitoring data from such programmes. During this study, flounder were collected from four separate sources; wild caught fish from the estuaries of the Rivers Alde, Mersey and Tyne, and farmed flounder from Port Erin Farm, Isle of Man. Under controlled laboratory conditions, groups of fish from each source were exposed to water-borne concentrations of the synthetic oestrogen ethynylestradiol (EE2) at a nominal concentration of 50 ng/l. Plasma was taken from each male fish after 6 and 10 days exposure and analysed for the presence of vitellogenin (VTG) using an ELISA technique. Significant levels of VTG induction were evident in fish from all sources after both 6 and 10 days exposure. Flounder from the Mersey were the only fish with significantly elevated initial background levels of VTG (day 0) and this appeared to be reflected in that these specimens showed the highest induction response after day 6. However, after day 10, fish from all other sites had a slightly higher mean VTG than those from the Mersey which showed significantly (p < 0.05) lower mean plasma VTG. It is suggested that other differential responses may have been masked by the use of a high dose of EE2 which produced maximum induction in nearly all fish. The findings of the study are discussed in terms of implications for further research into the differential response issue and how the initial plasma VTG figures contribute to a time-series from the Mersey, Tyne and Alde estuaries.     

Histopathological observations on liver,kidney and gonad of plaice (Pleuronectes platessa) taken from the Mersey estuary

The Mersey estuary is the most contaminated estuary in British waters. Detailed studies are underway on the pathology of flounder (Platichthys flesus) from the Mersey and on flounder from the nearby, but less contaminated, Dee estuary. Flounder breed offshore but spend a lot of time in the estuaries, penetrating into freshwater. These flatfish are in close contact with sediments and will be exposed to exenobiotic stored in the sediments. Plaice (Pleuronectes platessa) are flatfish which enter the Mersey with the tidal flow but spend less time in the estuary than flounder and are less likely to show evidence of pollutant exposure. Over 20 plaice were collected from the Mersey. Samples of the liver, kidney and gonad were fixed in 10% formal saline, processed into paraffin wax and 5 microm sections cut and stained with haematoxylin and eosin. Liver changes consisted of variable amounts of glycogen/lipid storage product, minor perivenular and perivascular fibrosis, and helminth parasitisation. No tumours or foci of cellular alteration, necrosis and inflammation were seen. In the gonad only changes consistent with normal gonadal cycling was seen. These studies provide evidence that plaice show little or no evidence of toxic pathological damage resulting from their stay in the estuary.     

Toxicopathic liver lesions in English sole and chemical contaminant exposure in Vancouver Harbour,Canada

The prevalence of toxicopathic liver lesions in English sole (Pleuronectes vetulus) was determined along a presumed gradient of chemical contamination in Vancouver Harbour, Canada. Fish were captured from five sites in or near Vancouver Harbour, British Columbia, Canada. No toxicopathic lesions were observed in fish examined at the reference site (Howe Sound outside Vancouver Harbour), or at the outer harbour site. In contrast, 20-23% of the fish from three sites located in the central harbour, Indian Arm and Port Moody Arm had one or more types of toxicopathic lesions. Likewise, aromatic hydrocarbon (AH) metabolites measured in bile exhibited a gradient in levels from lower concentrations at the reference site to significantly higher levels in fish from Indian Arm and Port Moody Arm harbour sites. The occurrence of toxicopathic liver lesions was statistically associated with concentrations of AHs measured in sediment and AH metabolite levels measured in bile.     

Liver histopathology in Baltic eelpout (Zoarces viviparus) – A baseline study for use in marine environmental monitoring

The use of liver histopathology in various fish species as a biomarker in marine monitoring programmes to assess the biological effect of exposure to anthropogenic contaminants is well established. However, although the eelpout (Zoarces viviparus) has repeatedly been recommended as an appropriate bioindicator for monitoring purposes in the Baltic Sea, information on histopathological liver alterations of this species is scarce. In order to obtain an overview, a total of 1070 eelpout, collected in the period between spring 2001 and winter 2010 from 34 sampling sites distributed over large parts of the Baltic Sea, were histologically analyzed. Liver lesions were recorded and classified according to established liver lesion categories. A high prevalence of non-specific and early toxicopathic non-neoplastic hepatic lesions was recorded. In contrast, pre-neoplastic lesions were observed only in rare cases and neoplastic changes were absent. A comparison with published data on other fish species revealed that the observed alterations may indicate effects of environmental stressors including contaminants. The results of the present study, thus, support the application of eelpout liver histopathology in environmental monitoring.     

Factors affecting the distribution of benthic macrofauna in estuaries contaminated by urban runoff

Contaminants derived from urban runoff have been shown to accumulate in estuarine sediments, reaching concentrations potentially capable of causing biological effects. Demonstration of effects, however, is difficult due to strong natural environmental gradients and the effects of past or present point-sources of contamination. We used multivariate methods to test two hypotheses relating to the effects of urban-derived contaminants on estuarine benthic communities. First, that patterns of distribution and abundance of benthic invertebrates in two urbanised estuaries would be different from those in two non-urbanised estuaries. Second, that the distributions of benthic invertebrates within and among the four estuaries would be related to those of urban-derived contaminants. Concentrations of contaminants were larger in estuaries with urbanised catchments and concentrations of Cu, Pb, Zn and DDT in some samples exceeded those at which biological effects may be expected to appear. Tests of differences in composition of benthic communities among estuaries showed that the two urban estuaries were not significantly different, but that they differed from both rural estuaries, which also differed from each other. Distributions of benthic invertebrates were significantly related to those of environmental variables, and were ordinated along axes that correlated with both natural environmental variables (nature of the sediment, position in estuary) and contaminants. Differences in faunas between the urban and non-urban estuaries were not, however, clear-cut and nor were relationships between faunal assemblages and environmental variables (including contaminants) consistent between two times of sampling.     

Growth of estuarine fish is associated with the combined concentration of sediment contaminants and shows no adaptation or acclimation to past conditions

We tested whether the growth rates of small benthic fish (Gillichthys mirabilis) in three southern California estuaries corresponded with the local concentrations of contaminants. Fish originating from each estuary were transplanted to cages in each estuary in two reciprocal transplant experiments. The growth rates of caged fish, and the size-distribution of natural populations, showed the same pattern of difference among estuaries. Twelve metals and organic contaminants occurred in bulk sediments at concentrations close to their individual ERL values, and a simple index of their combined concentration (the mean ERL quotient) was inversely correlated to the growth of caged fish. Metals in the water column occurred at lower concentrations, relative to toxicity thresholds, than those in sediments and were unrelated to fish growth. Fish used in the field caging experiments, and other fish held in the laboratory under constant conditions, showed no difference in growth according to their estuary of origin. Fish originating from different estuaries also showed no consistent differences in their tissue burden of organic contaminants. Our results thus suggested no genetic adaptation or physiological acclimation to the past contaminant regime, but revealed a possible association between fish growth rates and the combined concentration of multiple sediment contaminants.     

Estuarine and coastal connectivity of an estuarine-dependent fishery species,Pomadasys commersonnii (Haemulidae)

Understanding the level of connectivity between estuarine and coastal waters is essential for appropriate management of estuarine-associated taxa. Most studies have focused on the role of a single estuary, while limited research exists on the importance of multiple estuaries to individuals of estuarine-associated species. This study used acoustic telemetry to assess the usage of multiple estuaries and coastal waters by the estuarine-dependent spotted grunter Pomadasys commersonnii. Twenty-six adult fish were tagged with acoustic transmitters in the Kariega and Bushmans estuaries, South Africa, and their movements along a 300-km stretch of Indian Ocean coastline were monitored for up to 17 months. Tagged individuals spent most of their time in the estuary where they were tagged (55% and 85% for fish tagged in the two estuaries, respectively), followed by time in the sea (30% and 15%) and in other estuaries (15% and <1%). The mean durations of sea trips for fish tagged in the Kariega Estuary or Bushmans Estuary, respectively, were 25 days (range 3–55) and 12 days (range 2–22). Of the fish that went to sea, 93% from the Kariega Estuary and 60% from the Bushmans Estuary visited other estuaries. Most visits were undertaken to the Swartkops, Bushmans and Kowie estuaries, although the longest durations were spent in the Sundays Estuary. Individuals moved to estuaries up to 130 km away. The total distance travelled between estuaries by an individual during the study was ～529 km, with means of 201 and 184 km, respectively, for fish tagged in the Kariega and Bushmans estuaries. Despite covering large distances between estuaries, individuals often returned to their tagging estuary. Residency in their tagging estuary, combined with frequent visits to a neighbouring estuary, highlights the importance of estuarine habitats for this popular fish species, even after reaching maturity.     

The impact of river flooding and high flow on the demersal fish assemblages of the freshwater-dominated Great Fish Estuary,South Africa

Freshwater inflow has a strong impact on the biological, chemical and physical characteristics of estuaries, which in turn affect the distribution and abundance of estuarine organisms. Increased climatic variability associated with climate change is predicated to modify precipitation patterns, which will likely intensify floods in estuaries. The demersal fish assemblage of the freshwater-dominated Great Fish Estuary, South Africa, was sampled using beam trawls, monthly, from December 2013 to November 2014. The first six months of the study were characterised by river flooding and high flow, with estuarine conditions found only in the mouth region; this was followed by six months of intermediate flow, with estuarine conditions recorded up to 10 km from the mouth. River flooding and subsequent reduced salinity resulted in a decrease in species richness and abundances of fishes in the estuary, with only two estuarine species (Glossogobius callidus and Psammogobius knysnaensis) and one marine migrant (Solea turbynei) recorded following river flooding (201 m³ s^?1), in January 2014. The greatest species richness and abundances among both marine and estuarine fishes were recorded during intermediate flow conditions. We conclude that although freshwater inflow into estuaries is important for the nursery function of these systems, flooding—especially in freshwater-dominated estuaries—may cause a temporary decline in the abundance of most marine and estuarine fish species, including important bentho-pelagic marine migrant fishery species, such as Argyrosomus japonicus and Pomadasys commersonnii.     

Fish composition and assemblage structure in three Eastern English Channel macrotidal estuaries: A comparison with other French estuaries

This study has analysed for the first time fish composition and assemblage structures of three small macrotidal estuaries of the Eastern English Channel (EEC) and has explored the influences of 19 biotic and abiotic variables on the fish assemblages. Fish from Canche, Authie and Somme estuaries were collected during spring (June 2006 and May 2007) and autumn (September 2006) along the estuarine gradients using a 1.5 m beam trawl. Using identical sampling protocols, the study also analysed and compared for the first time taxonomic and functional aspects of the fish assemblages in 15 estuaries located along the Atlantic and English Channel coasts. SIMPER analysis showed high similarities in fish assemblages in the three EEC estuaries and during either spring or autumn periods. However, intra-estuary similarities were relatively low, indicating that fish assemblage structures (species richnesses or abundances) were more variable within the estuary (salinity gradient) than between estuaries and/or seasons (spring vs autumn). Although numerous environmental variables were included in the study, only 47% of the variability observed in the fish distribution was explained. Fish spatial variations in the EEC estuaries are mostly driven by abiotic variables as opposed to biological interactions. As indicated by CCA, salinity and muddy sediments were the two most important factors structuring the fish assemblages. The macrobenthos being very abundant in the EEC estuaries (580–1121 ind. m⁻²), the availability of potential prey is probably not a limiting factor in the utilization of estuaries by fish. Contrary to the majority of French estuaries dominated by estuarine species (ES), the fish assemblages of the EEC estuaries are clearly dominated by marine migrant (MM) species (65% on average) with high abundance of juveniles (mostly young-of-the-year). Cluster and SIMPROF's analyses distinguished the functional structure of the 15 estuarine fish assemblages into different clusters. The three EEC estuaries form a similar group with the largest French estuaries (Seine, Loire and Gironde). The latters were characterized by higher MM/ES ratio compared to the other French estuaries. Freshwater flow and tidal range were the main factors associated with this group of estuaries. The results of the present study do not indicate that anthropogenic impacts (chemical contamination) on estuaries affect their ecological functioning as described by the fish ecological guilds. It is suggested that the guild approach may not be useful to provide valuable information on the ecological status of estuaries.     

Assessing food web dynamics and relative importance of organic matter sources for fish species in two Portuguese estuaries: a stable isotope approach

Stable carbon and nitrogen isotopes (δ13C, δ1?N) were used to analyse food web dynamics of two of the main estuaries of the Portuguese coast: Tejo and Mira. The ultimate sources of organic matter supporting production of some of the most abundant and commercially important fish species were determined; and seasonal, inter- and intra- estuarine differences in the trophic relations among producers and consumers were identified. Stable isotope analysis was performed in different producers, primary consumers (main prey items for fish) and fish species (Solea solea, Solea senegalensis, Pomatoschistus microps, Dicentrarchus labrax, Liza ramada, Diplodus vulgaris and Atherina presbyter) of two areas in each estuary, in July and October 2009. Model calculations showed that the main prey for the fish species in the Tejo estuary used mostly salt marsh-derived organic matter as nutritional sources, with no marked differences between the sampled months. Trophic levels of fish species from the same estuary differed at multiple scales: inter-species, seasonally and spatially (both between and within estuaries). Significant differences in isotopic composition of fish species were more pronounced spatially (between the two sampled areas in the estuary) than seasonally (between sampled months). Trophic relationships in both estuaries demonstrated that organic matter is transferred to higher trophic positions mainly through benthic pathways. This shows the flexibility of these species to share resources and to exploit temporary peaks in prey populations. The present results showed that extensive disturbance in intertidal habitats from both estuaries may potentially change the balance of organic matter in the base of these complex food webs.     

Inter- and intra-estuarine fish assemblage variability patterns along the Portuguese coast

Estuaries along the Portuguese coast differ considerably in terms of their structure, geomorphologic and hydrologic characteristics. They play an important ecological role for different fish species, namely acting as important nursery areas. The fish assemblages of nine estuaries of the Portuguese coast were investigated in order to evaluate their main inter- and intra-estuarine variability patterns. Fish sampling surveys were conducted in May and July 2006, covering the full estuarine gradient. The different saline areas in each estuary were mapped using a Geographic Information System and fish assemblages’ were described and compared using a functional guilds approach. Generalized linear models were used to relate fish species richness to geomorphologic, hydrologic and environmental characteristics of the estuaries considered and correspondence analyses were performed to evaluate similarities in fish assemblages’ structure. At a large scale, river flow was the most important factor explaining the variability in species richness in estuaries along the Portuguese coast. At a regional scale, different abiotic factors explained the occurrence and abundance of fish species in the estuaries. Nonetheless, the overall role of the estuary was strongly related with the dominant saline zone within each estuary.     

Spatial variation in elemental composition of otoliths of three species of fish (family Sparidae)

Determining the nursery habitat of fishes that have moved from estuarine nursery habitats is difficult. The elemental fingerprints of otoliths of three species of sparids were determined to investigate their utility as a natural tag of the nursery habitat. Juvenile Pagrus auratus (snapper), Rhabdosargus sarba (tarwhine) and Acanthopagrus australis (bream) were collected from two sites in each of 15, six and three estuaries, respectively, and their otoliths analysed by solution-based inductively coupled plasma-mass spectrometry. Significant differences in otolith chemistry were found for all three species of juveniles collected from different estuaries. The same patterns among estuaries were not seen for all species, although it was not possible to sample the same sites within an estuary for all species. For bream, significant differences in otolith chemistry were found among all three estuaries, whereas for tarwhine the six estuaries were separated into three groups. For snapper, a number of estuaries could be separated, but there was some overlap for other estuaries. All three species were collected from the same site within one estuary and their otoliths analysed. Significant differences were found among species, but the implication of this finding remains unclear as the three species show differences in microhabitat use and may also differ in age. Because the elemental fingerprints of juveniles vary among estuaries or groups of estuaries, the nursery or recruitment estuary of adult fish could now be determined by analysing the juvenile region of adult otoliths. Thus, connectivity between estuaries and open coastal populations could be determined. Such information will have major implications for fisheries management because it will provide information on the distance that fish have moved from their recruitment estuary and the number of estuaries that contribute to each adult population.     

Choice chamber experiments to test the attraction of postflexion Rhabdosargus holubi larvae to water of estuarine and riverine origin

Although the recruitment of larvae and juveniles of marine fishes into estuaries has been well documented, little is known about the factors governing the immigration of estuary-associated marine fishes into estuaries. Fishes have a well-developed sense of smell and it has been suggested by several workers that olfactory cues of freshwater or estuarine origin serve as stimuli, attracting larvae and juveniles of estuary-associated species into estuaries. Attraction of postflexion Rhabdosargus holubi larvae to estuary and river water from the Kowie estuarine system, South Africa, was measured using a rectangular choice chamber. In experiments, conducted during peak recruitment periods, larvae selected estuary and river water with a significantly higher frequency than sea water. This study, the first to assess the possible role of olfaction in the recruitment process of an estuary-associated marine fish species, demonstrates that larvae are able to recognise water from different origins, probably based on odour.     

节点和岩岛对河口分汊的影响

本文主要论述作为先成边界条件的节点(包括矶)和岩岛对河口分汊的影响.指出节点和矶的分布和性质对河口能否形成分汊以及分汊形态等具有不容忽视的作用。表明单个节点易形成喇叭状的少汊河口,多个节点则易导致藕节状展宽及分汊河口,而节点位于口门,则不利于河口展宽而使得河口难以分汊。在研究河口岩岛对河口分汊的作用时,分别从珠江口、韩江口和闽江口三个分汊河口,分析了岩岛的数目多少、规模大小和分布状况,对导致河口形成不同分汊模式的影响。     

Artificial mouth opening fosters anoxic conditions that kill small estuarine fish

Fish kills are not uncommon within estuaries in many regions of the world. In seasonally open systems, which are common in temperate areas, they are often associated with mouth openings. Such a kill occurred in July 2005 in the Surrey Estuary following a closed mouth period of seven months resulting in the loss of many thousands of fish. At the time the fish community within the estuary was under investigation which provided comprehensive data of this population prior to the kill. Monthly water quality monitoring was also being conducted prior to the kill and also carried out on a daily basis following the mouth opening. The Surrey was stratified during the closed mouth phase, isolated waters below the halocline had stagnated and become anoxic. As a result only waters above the halocline contained oxygen concentrations capable of sustaining most fish. It appears that if a mouth opening happens under low flow conditions, a shearing effect occurs within the water column where surface waters flow out to sea leaving deeper waters behind. This resulted in only anoxic waters being present for in excess of six days and was responsible for the fish kill. Fish sampling of the Surrey Estuary was conducted three and six months following the kill and those data were compared to that collected in the 12 months prior to the event. Three months after the kill few fish were collected within the estuary and included marine opportunists near the mouth and estuarine resident species in the far upper reaches of the system. However six months following the kill large numbers of estuarine resident species were collected throughout the Surrey Estuary. As many species were euryhaline, it is believed that some individuals migrated into freshwater reaches of the Surrey to escape the anoxic conditions within the estuary. As conditions improved they recolonised the Surrey Estuary. The high fecundity and rapid growth of these small, short lived species probably aided in their re-establishing populations within the estuary. It is clear from this research that artificial openings of estuaries should be avoided during low flow periods when oxygen concentrations are low. It also appears that many of the small estuarine resident species common in seasonally open estuaries are capable of recolonising estuaries following fish kills. The effects on larger, longer lived resident species are not known but likely to be more detrimental due to longer time required for them to reach sexual maturity.     

Bioaccumulation and biomarkers in the sea star Asterias rubens (Echinodermata: Asteroidea): a North Sea field study

During two North Sea field trips in March 1995 and September 1996 sea stars, Asterias rubens, were collected at various stations along pollution gradients in order to study the relation between biochemical markers and levels of accumulated contaminants. Biomarkers measured were: cytochrome P450 level, benzo[a]pyrene hydroxylase (BPH) activity, acetylcholinesterase (AChE) activity and DNA integrity. Accumulation levels of heavy metals, polychlorinated biphenyls, organochlorine pesticides and polycyclic aromatic hydrocarbons (PAHs) in the pyloric caeca of sea stars indicate different pollution gradients, influenced by rivers in The Netherlands, UK and Germany. For some contaminants, especially PAHs, relatively high levels were found in the central part of the North Sea (Dogger Bank). On the basis of multivariate statistics, stations near the mouth of the Elbe and the Rhine/Meuse were shown to have different patterns of biomarker responses. Sea stars from stations in coastal zones showed relatively high levels of cytochrome P450 and 'P418', another haemoprotein that is present in most marine invertebrates. The station nearest to the Elbe Estuary showed the lowest BPH and AChE activity. DNA integrity was lower especially in stations near the Dutch coast and in a station near the Tees/Tyne estuaries. Using these biomarkers as early warning signals of exposure and/or adverse effects, this type of monitoring can be used also in the future to study the spatial and temporal trends in the quality of coastal waters.     

Variations of the Nutrients Loads to the Mainland U.K. Estuaries: Correlation with Catchment Areas, Urbanization and Coastal Eutrophication

The annual loads of nutrients (TO_xN equal to nitrate+nitrite; ammonium, phosphate, silicate) to all the estuaries on the mainland of the United Kingdom were estimated from data on water flow through gauging stations at the tidal limits of estuaries, and from concentration measurements under the Harmonised Monitoring Scheme of nutrient concentrations in water samples from these stations. The annual loads of nutrients showed distinct regional variations, with estuaries along the west coast of Wales and northern Scotland having much smaller loads than those along the east coast of England. The largest nitrogen loads were of TO_xN, and ammonium loads were usually small in comparison. The Severn, Mersey, Humber and Thames had the highest loads, although these were small in relation to the larger continental European estuaries. Loads of TO_xN per unit of catchment area were surprisingly constant (about 10⁵ moles N km⁻² y⁻¹). The nutrient loads showed that most U.K. catchments were influenced by human activity, the majority being in the ‘ moderately influenced ’ category. Nutrient loads were also normalized for the area of each estuary, as a measure of the relative influence of nutrients on the receiving estuaries. The ratios of N:P, N:Si and P:Si in the annual loads suggested that most estuaries were likely to be, if anything, P limited rather than N or Si limited. However, crude annual loads may conceal significant seasonal variations.The spring maximum chlorophylla concentrations in coastal waters adjacent to each estuary were significantly correlated with the log total annual loads of TO_xN, ammonium and phosphate (but not silicate) for each estuary, providing a direct link between a measure of the degree of biological response in coastal waters and the nutrient load through the estuaries. There were no significant correlations between spring maximum chlorophyll a concentrations and either catchment-normalized or estuary-normalized nutrient loads. There was significant correlation between catchment area-normalized loads of phosphate and an urbanization index for the catchments, but not with the catchment area-normalized loads of the other nutrients.