Environmental pollution and natural populations: a biomarkers case study from the Iberian Atlantic coast

The degradation of estuaries is a result of human activities which overloads the environment with substances of both industrial and/or natural origins. Bioindicators have been consistently used to interpret effects of contaminants in the environment. In this study, the use of biomarkers (particular measurable characteristics of a bioindicator organism) was used to evaluate the contamination by xenobiotics of Crangon crangon natural populations. The central aim was to evaluate the capability of a battery of biomarkers to discriminate sites with different types of contamination. The activity of the enzymes cholinesterases (ChE), lactate dehydrogenase (LDH) and glutathione S-transferases (GST) were used as biomarkers. In addition, the ChE form(s) present in the cephalotorax of C. crangon were characterised. Organisms were seasonally sampled from winter 2001/2002 to autumn of 2002, at "reference" sites and at sites that receive agricultural, industrial and/or urban effluents. Results obtained in the characterisation of ChE with different substrates and selective inhibitors demonstrate that the form of ChE present in the cephalotorax of C. crangon shows proprieties of vertebrates' AChE and therefore it may be classified as true AChE-like ChE. The battery of biomarkers exhibited seasonal and local variations, apparently related to agricultural, industrial or urban effluent contamination. The tested biomarkers proved to be able to discriminate sources of environmental contamination, and confirms C. crangon as a sensitive species suitable to be used as a bioindicator.     

First evidence of altered vitellogenin-like protein levels in clam Tapes philippinarum and in cockle Cerastoderma glaucum from the Lagoon of Venice

Possible xenoestrogenic effects were investigated, for the first time, in two bivalve species from the Lagoon of Venice (Italy): the clam Tapes philippinarum and the cockle Cerastoderma glaucum. Bivalves were collected far from their reproductive phase at the very early stage of gametogenesis (January), and in the pre-spawning period (June) in six sites. Vitellogenin (Vg)-like proteins (a biomarker of exposure to estrogenic compounds) were measured by the alkali-labile phosphate method (ALP), in both haemolymph and digestive gland from males and females (when it was possible to distinguish sex by microscopic observation of gonadal tissue). Haemolymph calcium concentrations (a parameter considered closely related to the presence of Vg-like proteins) and the bivalve condition index (CI) were also measured. In both seasons, bivalves collected at Campalto (near a sewage treatment plant) and Marghera (a highly contaminated area) had higher Vg-like protein levels, particularly in haemolymph, than animals from the other sampling sites. Interestingly, CI had high values in these polluted sites. In June only, Vg-like proteins and Ca(2+) levels in haemolymph exhibited similar trends in both bivalve species at most sampling sites. The responsiveness of bivalves to environmental xenoestrogens was higher in June, allowing better discrimination among sites. The present study demonstrates that animals from highly polluted areas have increased Vg-like protein levels. As endocrine disruption due to exposure to estrogenic compounds may cause fertility reduction, alterations in the sex ratio, and a decrease in reproductive rate, a condition of potential risk for bivalve populations in estuarine areas is highlighted.     

First evidence of altered vitellogenin-like protein levels in clam Tapes philippinarum and in cockle Cerastoderma glaucum from the Lagoon of Venice

Possible xenoestrogenic effects were investigated, for the first time, in two bivalve species from the Lagoon of Venice (Italy): the clam Tapes philippinarum and the cockle Cerastoderma glaucum. Bivalves were collected far from their reproductive phase at the very early stage of gametogenesis (January), and in the pre-spawning period (June) in six sites. Vitellogenin (Vg)-like proteins (a biomarker of exposure to estrogenic compounds) were measured by the alkali-labile phosphate method (ALP), in both haemolymph and digestive gland from males and females (when it was possible to distinguish sex by microscopic observation of gonadal tissue). Haemolymph calcium concentrations (a parameter considered closely related to the presence of Vg-like proteins) and the bivalve condition index (CI) were also measured. In both seasons, bivalves collected at Campalto (near a sewage treatment plant) and Marghera (a highly contaminated area) had higher Vg-like protein levels, particularly in haemolymph, than animals from the other sampling sites. Interestingly, CI had high values in these polluted sites. In June only, Vg-like proteins and Ca²⁺ levels in haemolymph exhibited similar trends in both bivalve species at most sampling sites. The responsiveness of bivalves to environmental xenoestrogens was higher in June, allowing better discrimination among sites. The present study demonstrates that animals from highly polluted areas have increased Vg-like protein levels. As endocrine disruption due to exposure to estrogenic compounds may cause fertility reduction, alterations in the sex ratio, and a decrease in reproductive rate, a condition of potential risk for bivalve populations in estuarine areas is highlighted.     

First evidence of altered vitellogenin-like protein levels in clam Tapes philippinarum and in cockle Cerastoderma glaucum from the Lagoon of Venice

Possible xenoestrogenic effects were investigated, for the first time, in two bivalve species from the Lagoon of Venice (Italy): the clam Tapes philippinarum and the cockle Cerastoderma glaucum. Bivalves were collected far from their reproductive phase at the very early stage of gametogenesis (January), and in the pre-spawning period (June) in six sites. Vitellogenin (Vg)-like proteins (a biomarker of exposure to estrogenic compounds) were measured by the alkali-labile phosphate method (ALP), in both haemolymph and digestive gland from males and females (when it was possible to distinguish sex by microscopic observation of gonadal tissue). Haemolymph calcium concentrations (a parameter considered closely related to the presence of Vg-like proteins) and the bivalve condition index (CI) were also measured. In both seasons, bivalves collected at Campalto (near a sewage treatment plant) and Marghera (a highly contaminated area) had higher Vg-like protein levels, particularly in haemolymph, than animals from the other sampling sites. Interestingly, CI had high values in these polluted sites. In June only, Vg-like proteins and Ca²⁺ levels in haemolymph exhibited similar trends in both bivalve species at most sampling sites. The responsiveness of bivalves to environmental xenoestrogens was higher in June, allowing better discrimination among sites. The present study demonstrates that animals from highly polluted areas have increased Vg-like protein levels. As endocrine disruption due to exposure to estrogenic compounds may cause fertility reduction, alterations in the sex ratio, and a decrease in reproductive rate, a condition of potential risk for bivalve populations in estuarine areas is highlighted.     

Environmental management in Antarctica Instruments and institutions

Instruments and institutions for environmental management in Antarctica have evolved from beginnings in the Antarctic Treaty, with subsequent introduction of the Convention for the Conservation of Antarctic Seals, the Convention on the Conservation of Antarctic Marine Living Resources, and recommendations made at Treaty meetings. The Protocol on Environmental Protection to the Antarctic Treaty will improve existing provisions for environmental management in Antarctica. When it enters into force the Protocol will apply to all activities in the Treaty area: it provides rules for environmental impact assessment, conservation of fauna and flora, disposal of wastes, prevention of marine pollution and specially protected and managed areas. The Protocol's Committee for Environmental Protection will facilitate discussion and advice on environmental management. Management of information is a vital part of ensuring measures are effective. The Protocol should be ratified quickly to improve the basis for environmental protection, but the test will be whether and how it is implemented in practice.     

Biochemical and microbial features of shallow marine sediments along the Terra Nova Bay (Ross Sea,Antarctica)

Shallow marine sediments were collected from seven stations (three of which located at Gerlache Inlet, two at Tethys Bay, one at Adelie Cove and one just beneath the Italian Research Base) along the Terra Nova Bay coast (Ross Sea, Antarctica). Their chemical, biochemical and microbiological properties were studied in order to provide further insights in the knowledge of this Antarctic benthic ecosystem. Overall, the organic carbon (OC) represented the major fraction of total carbon (TC) and displayed concentrations similar to or slightly lower than those previously measured in Antarctic bottom sediments. The biopolymeric carbon within OC ranged from 4.1% to 19.9% and showed a wide trophic range (65–834 μg g⁻¹ d.w.). Proteins (PRT) represented on average the main biochemical class contributing to labile organic carbon, followed by lipids (LIP) and carbohydrates (CHO). The activity of aminopeptidase, β-d-glucosidase, alkaline phosphatase and esterase was checked, giving the highest values at Tethys Bay and at the deepest water sediments. The principal component analysis, which was computed considering physical, chemical (elemental and biochemical sedimentary composition) and microbiological parameters (including bacterial abundance, ectoenzymatic activities, T-RFs richness and diversity indices), allowed to obtain two main clusters (“Tethys Bay” and “other stations”). Based on data obtained, two representative 16S rRNA clone libraries using samples from Tethys Bay and Gerlache Inlet were constructed. The sequences of 171 clones were compared to those available in public databases to determine their approximate phylogenetic affiliations. Both aerobic and anaerobic bacteria were disclosed, with the majority of them affiliated with the Gamma- and Deltaproteobacteria, Bacteroidetes and Acidobacteria. The occurrence of strictly anaerobic bacteria suggests that sediments might also undergo anoxic conditions that, in turn, could favor the accumulation of PRT in respect to CHO, assuming that fermentation of amino acids is slower than that of sugars from decomposing polysaccharides.     

Marine introductions in the Southern Ocean: an unrecognised hazard to biodiversity

This study investigated the potential for transport of organisms between Hobart, Macquarie Island and the Antarctic continent by ships used in support of Antarctic science and tourism. Northward transport of plankton in ballast water is more likely than southward transport because ballast is normally loaded in the Antarctic and unloaded at the home port. Culturing of ballast water samples revealed that high-latitude hitchhikers were able to reach greater diversities when cultured at temperate thermal conditions than at typical Southern Ocean temperatures, suggesting the potential for establishment in the Tasmanian coastal environment. Several known invasive species were identified among fouling communities on the hulls of vessels that travel between Hobart and the Southern Ocean. Southward transport of hull fouling species is more likely than northward transport due to the accumulation of assemblages during the winter period spent in the home port of Hobart. This study does not prove that non-indigenous marine species have, or will be, transported and established as a consequence of Antarctic shipping but illustrates that the potential exists. Awareness of the potential risk and simple changes to operating procedures may reduce the chance of introductions in the future.     

Physiological and genetical adaptation to temperature in fish populations

The physiological and genetical adaptation of fishes to environmental temperatures has been studied by analyzing data concerning: (i) the oxygen binding properties of haemoglobin recorded during growth experiments on Atlantic cod, and (ii) the primary structure of haemoglobin (Hb) and lactate dehydrogenase (LDH) of several fish species living in polar and temperate areas. The results on the oxygen binding properties of cod's haemoglobin indicate that for this species a temperature of around 12°C is the most favourable one, irrespective of the haemoglobin genotype, and are in line with recent evidence challenging the existence of significant evolutionary differences between cod stocks in North Atlantic. The primary structures of both Hb and LDH from species living under temperate environments show a higher variability as compared to that from polar species, although the difference in the recurrent patterns of hydrophobicity between the two areas is much larger for Hb. These results highlight the dominant role of physiological and genetical factors in shaping the adaptation to temperature at the individual and at the species level, respectively.     

Assessment of the macrobenthos in an Antarctic environment following recent pollution abatement

Quantative population data for shallow Antarctic soft-bottom communities in King Edward Cove, South Georgia indicate that although once grossly polluted by organic effluent and fuel oil from a whaling station, the fauna recovered within some eight years after contamination ceased. The results are discussed in relation to comparable studies in other polar and temperate areas.     

Monitoring of DSP toxins in small-sized plankton fraction of seawater collected in Mutsu Bay,Japan, by ELISA method: relation with toxin contamination of scallop

Monitorings were conducted on DSP toxins in mid-gut gland of scallop (mouse assay), cell numbers of toxic dinoflagellate species of Dinophysis, and diarrhetic shellfish poisoning (DSP) toxins in small-sized (0.7-5 microm) plankton fraction of seawater collected from surface (0 m) and 20 m depth at a station in Mutsu Bay, Aomori Prefecture, Japan, in 2000. A specific enzyme-linked immunosorbent assay (ELISA) was employed for the analysis of DSP toxins in small-sized plankton fraction using a mouse monoclonal anti-okadaic acid antibody which recognizes okadaic acid, dinophysistoxin-1, and dinophysistoxin-3. DSP toxins were detected twice in the mid-gut gland of scallops at 1.1-2.3 MU (mouse units) g(-1) on 26 June and at 0.6-1.2 MU g(-1) on 3 July, respectively. Relatively high cell densities of D. fortii were observed on 26 June and 11 September, and may only contribute to the bivalve toxicity during late June to early July. D. acuminata did not appear to be responsible for the toxicity of scallops in Mutsu Bay in 2000. ELISA monitoring of small-sized plankton fraction in seawater could detect DSP toxins two weeks before the detection of the toxin in scallops, and could do so two weeks after the loss of the bivalve toxicity by mouse assay. On 17 July, toxic D. fortii was detected at only small number, <10 cells l(-1), but DSP toxins were detected by the ELISA assay, suggesting a presence of other toxic small-sized plankton in seawater. For the purpose of reducing negative impacts of DSP occurrences, monitorings have been carried out hitherto on DSP toxins of bivalve tissues by mouse assay and on cell densities of "toxic" species of Dinophysis. Here we propose a usefulness of ELISA monitoring of plankton toxicity, especially in small-sized fraction, which are possible foods of mixotrophic Dinophysis, as a practical tool for detecting and predicting DSPs in coastal areas of fisheries grounds of bivalve aquaculture.     

Natural water-purification system observed in a shallow coastal lagoon: Matsukawa-ura,Japan

Field surveys and in situ experiments were conducted in the shallow Matsukawa-ura in summer to evaluate the biological efficiencies of shallow-water areas for preserving coastal ecosystems. In Matsukawa-ura (5.8 km(2)), the suspension-feeding bivalves Ruditapes philippinarum and Crassostrea gigas were the dominant animals--their total biomasses (wet weight) were estimated to be 3.4 x 10(6) and 2.3 x 10(6) kg, respectively. Ulva sp. and Zostera marina were the dominant macrophyte species during the summer, with standing crops estimated to be 0.29 x 10(6) and 0.20 x 10(6) kg, respectively. The dissolved inorganic nitrogen (DIN) uptake rates for Ulva sp. and Z. marina were determined by in situ experiments. An ecological model calculated on the basis of the observed dataset showed that, in comparison with tidal exchange, a significant amount of particulate organic matter was removed by bivalve filtration and a considerable quantity of DIN was removed by macrophyte species.     

Benthic marine pollution around McMurdo Station, Antarctica: A summary of findings

McMurdo Station is the United States Antarctic Research Program's (USARP) centre for scientific and logistical support for most of the continent and is the largest settlement in Antarctica. Over 30 years of human presence with shipping, air-transport, scientific, and municipal activities have resulted in severe but localized nearshore marine benthic pollution. This review will summarize the results of 3 years of research which has documented concentrations of chemical contaminants, changes in community patterns, and the toxicity of sediments to invertebrate species and infaunal communities. The primary contaminants are petroleum hydrocarbons in the sediments of Winter Quarters Bay, the site of shipping activities and a former dumpsite. Total hydrocarbon levels in sediments in Winter Quarters Bay were comparable to the most polluted harbours in temperate latitudes. However, a steep gradient of pollution existed: stations less than 1 km away were nearly pristine. Benthic invertebrate communities changed dramatically along this contamination gradient. The results of both field and laboratory bioassay experiments with contaminated sediments found that biological changes observed in benthic communities around McMurdo Station were most likely caused by hydrocarbons, PCBs, and PCTs. A primary goal of polar pollution research is to determine the response and tolerance of Antarctic biota to physical and chemical anthropogenic disturbance, and the mitigation of negative effects. To this end, future work should include continued monitoring of changes in chemical and community patterns, studies of the physiological responses of Antarctic invertebrates to chemical contamination, and further comparisons of anthropogenic and natural disturbances to benthic communities.     

Snowdrift effect on snow deposition: Insights from a comparison of a snow pit profile and meteorological observations in east Antarctica

A high-frequency and precise ultrasonic sounder was used to monitor precipitated/deposited and drift snow events over a 3-year period(17 January 2005 to 4 January 2008) at the Eagle automatic weather station site,inland Antarctica.Ion species and oxygen isotope ratios were also generated from a snow pit below the sensor.These accumulation and snowdrift events were used to examine the synchronism with seasonal variations of δ~(18)O and ion species,providing an opportunity to assess the snowdrift effect in typical Antarctic inland conditions.There were up to 1-year differences for this 3-year-long snow pit between the traditional dating method and ultrasonic records.This difference implies that in areas with low accumulation or high wind,the snowdrift effect can induce abnormal disturbances on snow deposition.The snowdrift effect should be seriously taken into account for high-resolution dating of ice cores and estimation of surface mass balance,especially when the morphology of most Antarctic inland areas is similar to that of the Eagle site.     

A multibiomarker approach in barramundi (Lates calcarifer) to measure exposure to contaminants in estuaries of tropical North Queensland

A suite of biomarkers were measured in barramundi (Lates calcarifer) from five North Queensland estuaries along a perceived pollution gradient. The biomarkers selected were 7-ethoxyresorufin-O-deethylase (EROD), cytochrome P450, fluorescent aromatic compounds (FACs), DNA integrity, RNA:DNA ratio, cholinesterase activity (ChE), condition factor and hepatosomatic index. The resulting database was subjected to uni- and multi-variate analyses in order to assess the most suitable biomarkers to assess pollution in North Queensland estuaries and to classify the environmental quality of the sites. Principal components analysis (PCA) on the biochemical markers revealed that EROD, EROD/P450, DNA damage and to a lesser extent ChE and FACs were found to be responsive to contaminants in the environment while cytochrome P450, condition factor and the hepatosomatic index were found to be less responsive biomarkers. This study has demonstrated the utility of applying a multibiomarker approach in conjunction with traditional analysis of contaminants in providing valuable information in environmental risk assessment.     

Evaluating the genotoxic damage and hepatic tissue alterations in demersal fish species: a case study in the Ligurian Sea (NW-Mediterranean)

A protocol for detecting hepatic micronuclei in fish was performed to check genotoxic damage, as an indicator of environmental hydrocarbons exposure, in relation to the "Haven" oil spill. As target fish, we have chosen three demersal species with different habitats and feeding behaviour (i.e., Lepidorhombus boscii, Merluccius merluccius and Mullus barbatus) collected from two differently impacted areas and a control site. Additional analysis was performed by histological detection of hepatic tissue damages such as the presence of necrotic and tumour-like aspects. The three studied species showed different sensitivity to environmental pollutants exposure, L. boscii resulting the more sensitive in terms of both micronuclei incidence and tissue damage. The results of this study show that: (1) the micronucleus test could be an effective and fast method to detect oil pollution; (2) a clear response of L. boscii only to oil contamination for both micronucleus test and liver tissue alterations.     

Delta13C and delta15N shifts in benthic invertebrates exposed to sewage from McMurdo Station, Antarctica

In an effort to identify biomonitors for contamination of Antarctic marine benthos by sewage, this study determines whether the US Antarctic Program’s McMurdo Station produces a benthic sewage footprint and whether resident megafauna are assimilating sewage-derived material. We identified strong C and N isotopic gradients in benthic sediment as a function of downstream distance from McMurdo Station’s point-source sewage addition. Sediment C and N isotope ratios approached marine background levels at the sampling end-point 612 m downcurrent. Based on isotope abundances in their tissues, at least some sewage C and N were assimilated by the sedentary, suspension feeding soft coral Alcyonium antarcticum, ascidian Cnemidocarpa verrucosa and bivalve Laternula elliptica. However, as inferred by tissue-sediment differences in downstream isotope trends, such assimilation was not in proportion to sewage exposure and input, therefore implying non-generalist feeding behavior by these species. In contrast, the motile, generalist feeding sea urchin Sterechinus neumayeri, sea star Odontaster validus and ribbon worm Parborlasia corrugatus showed isotopic evidence of sewage C and N assimilation roughly in proportion to sewage input. We recommend these generalist feeders for further use as biomonitors at this site now that sewage treatment has been implemented. As these species are circumpolar in distribution, they may also prove useful elsewhere in the Antarctic.     

Coprostanols in Antarctic marine sediments: A biomarker for marine mammals and not human pollution

The first analysis of sterols in Antarctic marine sediment cores (Bransfield Strait and McMurdo Sound) is reported here. The isolated sterols in the sediments from Bransfield Strait are comprised mainly of dinosterol (4,23,24-trimethyl-5cholest-22-en-3β-ol), coprostanol (5β-cholestan-3β-ol) and epicoprostanol (5β-cholestan-3-ol) and trace amounts of cholesterol (cholest-5-en-3β-ol). The sediment cores of McMurdo Sound also contain small quantities of coprostanols in addition to large amounts of marine sterols. Dinosterol is a known dinoflagellate chemical marker, whereas coprostanols are currently being used as tracers for sewage contamination. However, sterol composition in Antarctic sediments is expected to be atypical of sewage input and probably different from those of temperate and tropical zones. The coprostanols must therefore arise from the feces of marine mammals, most probably whales which use the region for feeding and spawning. Based on the results of this study, caution should be exercised in the use of coprostanol to estimate the amount of sewage-derived organic matter transported away from impacted coastal areas.     

Environmental assessment of a large industrial marine complex based on a community of benthic filter-feeders

Biological quality in a bay affected by man's activities was evaluated by means of the composition of assemblages of sponges and ascidians. Our results showed that the structure of these two groups of filter-feeders aided in discriminating between undisturbed and disturbed areas, establishing different "environmental health categories" from moderately to strongly disturbed areas, and in ascertaining the extension of the area of each "health category". We were able to divide the bay into four zones based on type of disturbance or anthropogenic source: (1) stations free of any source of disturbance, (2) stations under moderate disturbance, located close to industrial ports, millworks, etc., (3) stations that are under the direct influence of industrial wastes such as a power station and oil refinery, and (4) stations near strongly disturbed areas, influenced directly by harmful steelworks activities. We differentiated clearly between four large species assemblages, and related the composition of these assemblages to different kinds of disturbances. Thus, these species could be used to manage the marine environment in this bay by comparing the observed fauna, with expected fauna in an unstressed site. Moreover, the joint presence of the sponge Cliona vastifica and tunicate Policitor adriaticum seems always to indicate a more or less pristine environmental situation, functioning as bioindicators of normal conditions. We think that the use of specific bioindicators for monitoring disturbance is a valid tool to establish baselines to predict impacts associated with industrial development in many marine ecosystems. The advantages to monitoring communities on hard rocks versus sandy or muddy bottoms are also commented upon.     

Assessing the relative role of climate change and human activities in sandy desertification of Ordos region,China

Climate change and human activities are driving forces of sandy desertification and the relative role of them in sandy desertification is the hot point in related researches. A study was carried to assess the relative role of climate change and human activities in sandy desertification of Ordos region in China. Potential NPP and the difference between potential and actual NPP were selected as indicators to assess the relative role of climate change and human activities in sandy desertification, respectively. Assessing methods were built based on some scenarios for the reversion and expansion of sandy desertification and the relative role of climate change and human activities in sandy desertification of Ordos region were assessed from 1981 to 2000. The results showed that although some local places experienced an expansion of sandy desertification, the change of sandy desertification of Ordos region from 1981 to 2000 showed a stably reversing trend. The relative role of climate change and human activities in sandy desertification of Ordos region varied at different temporal and spatial scales in the reversion and expansion processes. In the reversion of sandy desertification, climate change was the dominant factor in the period of 1981 to 1990 and the reversed areas mainly induced by climate change including Mu Us sandy land and the transition zone between temperate steppe and temperate deciduous scrubs in north east of Ordos region; however, human activities controlled the reversed process during the period from 1991 to 2000 and the areas mainly induced by human activities distributed in all banners of Ordos region. In the expansion of sandy desertification, human activities were the dominant factor in the period of 1981 to 1990 and the expanded areas mainly included the regions around common boundary of Hanggin, Dalad Banners and Dongsheng City; however, climate change dominated the expansion of sandy desertification from 1991 to 2000, the expanded areas equably distributed in the whole of Ordos region except the south east of Otog Qian, Uxin and Jungar Banners. Supported by Chinese National Academic Institutional Commonweal Research Programs (Grant No. 2005DIA3J006)     

In situ enzyme activities of filamentous scum bacteria in municipal activated sludge wastewater treatment plants

In situ phosphatase, esterase, lipase, and β‐glucuronidase activities were investigated in filamentous scum bacteria by ELF (enzyme labeled fluorescence)‐technology. Microthrix parvicella exhibited significant activities for all four enzymes with lipase activity being the highest. In situ activities were considerably higher in activated sludge as compared to scum indicating M. parvicella growth to occur mainly in the sludge fraction. “Nocardioform actinomycetes” showed significant activities for phosphatase, esterase, and β‐glucuronidase, lipase activity was only moderate. Activities revealed to be similar for activated sludge and scum. As population densities of enzyme‐active actinomycetes were noticeably higher in scum they presumably find good growth conditions in the scum layer. Enzyme activities in Nostocoida limicola morphotypes were only low with no lipase activity being detected. Due to their phylogenetic diversity activity assignment should be combined with fluorescence in situ hybridization (FISH). Numerous type 0041/0675 and type 1851 filaments showed all but lipase activity. However, some of their attached bacteria revealed to be lipase‐active. Only a few morphotype 0092 filaments revealed phosphatase, esterase, and β‐glucuronidase activity. ELF investigations proved suitable for monitoring in situ filamentous activity. Present imponderabilities of Eikelboom morphotype phylogenetic affiliation are discussed.