Harm G. van der Geest

Comparative ecotoxicity of NPAHs to larvae of the midge Chironomus riparius

Abstract Acute toxicity of seven azaarenes (NPAHs) has been determined using larvae of the midge Chironomus riparius. Clear dose-response relationships for mortality of first instar larvae were observed, but surviving larvae grew as equally well as the controls. LC50 values (96 h) for a two-ringed structure (quinoline), four three-ringed structures (acridine, phenanthridine, benzo[f]quinoline and benzo[h]quinoline) and two four-ringed structures (benz[a]acridine and benz[c]acridine) were generally one order lower than the few values reported for other aquatic species in the literature. Dibenz[a,i]acridine, a five-ringed structure, was not acutely toxic at its maximum dissolved concentration. Acute toxicity increased with increasing number of aromatic rings of the compound; accordingly, correlations between the LC50 values and size, shape and topology-related molecular properties were strong. However, between the four benzoquinoline isomers tested also differences in toxicity were observed: acridine was significantly more toxic than the other benzoquinoline isomers tested. This is most likely caused by transformation of acridine under UV light, indicated by a HOMO–LUMO gap in the highly phototoxic region, a high correlation between isomer toxicity and heat of formation and a low recovery of acridine in the water during the experiment.

Networking our way to better ecosystem service provision

The ecosystem services (EcoS) concept is being used increasingly to attach values to natural systems and the multiple benefits they provide to human societies. Ecosystem processes or functions only become EcoS if they are shown to have social and/or economic value. This should assure an explicit connection between the natural and social sciences, but EcoS approaches have been criticized for retaining little natural science. Preserving the natural, ecological science context within EcoS research is challenging because the multiple disciplines involved have very different traditions and vocabularies (common-language challenge) and span many organizational levels and temporal and spatial scales (scale challenge) that define the relevant interacting entities (interaction challenge). We propose a network-based approach to transcend these discipline challenges and place the natural science context at the heart of EcoS research.

Cell Turnover and Detritus Production in Marine Sponges from Tropical and Temperate Benthic Ecosystems

This study describes in vivo cell turnover (the balance between cell proliferation and cell loss) in eight marine sponge species from tropical coral reef, mangrove and temperate Mediterranean reef ecosystems. Cell proliferation was determined through the incorporation of 5-bromo-2′-deoxyuridine (BrdU) and measuring the percentage of BrdU-positive cells after 6 h of continuous labeling (10 h for Chondrosia reniformis). Apoptosis was identified using an antibody against active caspase-3. Cell loss through shedding was studied quantitatively by collecting and weighing sponge-expelled detritus and qualitatively by light microscopy of sponge tissue and detritus. All species investigated displayed substantial cell proliferation, predominantly in the choanoderm, but also in the mesohyl. The majority of coral reef species (five) showed between 16.1±15.9% and 19.0±2.0% choanocyte proliferation (mean±SD) after 6 h and the Mediterranean species, C. reniformis, showed 16.6±3.2% after 10 h BrdU-labeling. Monanchora arbuscula showed lower choanocyte proliferation (8.1±3.7%), whereas the mangrove species Mycale microsigmatosa showed relatively higher levels of choanocyte proliferation (70.5±6.6%). Choanocyte proliferation in Haliclona vansoesti was variable (2.8–73.1%). Apoptosis was negligible and not the primary mechanism of cell loss involved in cell turnover. All species investigated produced significant amounts of detritus (2.5–18% detritus bodyweight−1·d−1) and cell shedding was observed in seven out of eight species. The amount of shed cells observed in histological sections may be related to differences in residence time of detritus within canals. Detritus production could not be directly linked to cell shedding due to the degraded nature of expelled cellular debris. We have demonstrated that under steady-state conditions, cell turnover through cell proliferation and cell shedding are common processes to maintain tissue homeostasis in a variety of sponge species from different ecosystems. Cell turnover is hypothesized to be the main underlying mechanism producing sponge-derived detritus, a major trophic resource transferred through sponges in benthic ecosystems, such as coral reefs.

Invertebrate footprints on detritus processing, bacterial community structure, and spatiotemporal redox profiles

Abstract.  Detritus processing is driven by a complex interplay between macroinvertebrate and microbial activities. Bioturbation/feeding activities of invertebrates in sediments are known to influence decomposition rates. However, direct effects of invertebrates on bacterial communities and detritus processing remain ill-defined, mainly because identifying interactions between invertebrates and sediments is methodologically challenging. We incubated 5 macroinvertebrate species with various bioturbation/feeding traits separately in sediment-filled microcosms inoculated with bacterial communities for 5 d. At the end of the experiment, we assessed: 1) detritus processing (mass loss on ignition [LOI] and dissolved organic C accumulation in the overlying water [absorbance at 280 nm]), 2) bacterial community structure (intergenic spacer analysis [RISA]) and bacterial activity (electron transport system activity [ETSA]), and 3) development of redox potential (Eh) over time (with permanently installed microelectrodes). Invertebrates enhanced bacterial activity and detritus processing, and the magnitude depended on bioturbation/feeding traits. Bacterial community structure differed significantly between microcosms with burrowing invertebrates and microcosms with sediment-dwelling invertebrates. Eh profiles were similar among microcosms with invertebrates with similar bioturbation/feeding traits, but differed among microcosms with invertebrates with dissimilar bioturbation/feeding traits. Our results suggest that bioturbation by aquatic invertebrates mediates detritus processing, Eh dynamics, and structure of the microbial community. These findings highlight the significance of bioturbation and show the utility of spatiotemporal Eh dynamics as footprints reflecting functioning of benthic detrital food webs.

Seasonal variability in irradiance affects herbicide toxicity to the marine flagellate Dunaliella tertiolecta

Photosynthetically Active Radiation (PAR) and Ultraviolet Radiation (UVR) of the solar spectrum affect microalgae directly and modify the toxicity of phytotoxic compounds present in water. As a consequence seasonal variable PAR and UVR levels are likely to modulate the toxic pressure of contaminants in the field. Therefore the present study aimed to determine the toxicity of two model contaminants, the herbicides diuron and Irgarol®1051, under simulated irradiance conditions mimicking different seasons. Irradiance conditions of spring and autumn were simulated with a set of Light Emitting Diodes (LEDs). Toxicity of both herbicides was measured individually and in a mixture by determining the inhibition of photosystem II efficiency (ΦPSII) of the marine flagellate Dunaliella teriolecta using Pulse Amplitude Modulation (PAM) fluorometry. Toxicity of the single herbicides was higher under simulated spring irradiance than under autumn irradiance and this effect was also observed for mixtures of the herbicides. This irradiance dependent toxicity indicates that herbicide toxicity in the field is seasonally variable. Consequently toxicity tests under standard light conditions may overestimate or underestimate the toxic effect of phytotoxic compounds.

Mangrove-sponge associations: a possible role for tannins

A positive correlation between sponge coverage and tannin concentrations in prop roots of Rhizophora mangle L. has previously been reported. However, the ecological role of tannins within the mangrove sponge association remains speculative. This study investigated whether tannins play a role in sponge recruitment and assessed tannin and polyphenol production in R. mangle roots in response to sponge colonization. We demonstrated in a field experiment using artificial substrates with different tannin concentrations that tannins are positively involved in larval recruitment of the sponge Tedania ignis and that roots significantly enhanced tannin and polyphenolic content in response to natural and experimental sponge fouling. Differential recruitment in response to tannins may have been the result of a behavioral response in sponge larvae. It is also possible that tannins affected the structure of the fouling microbial biofilm on the artificial substrate, or tannins affected the post-settlement dynamics of sponge recruits. Elevations in concentrations of tannins and polyphenolic compounds upon coverage with sponges, combined with differential recruitment of T. ignis in response to differences in tannin concentrations, may indicate a positive feedback in recruitment. This may in part explain the typical heterogeneity in sponge coverage and community composition among roots.

Resource niche overlap promotes stability of bacterial community metabolism in experimental microcosms

Decomposition of organic matter is an important ecosystem process governed in part by bacteria. The process of decomposition is expected to benefit from interspecific bacterial interactions such as resource partitioning and facilitation. However, the relative importance of resource niche breadth (metabolic diversity) and resource niche overlap (functional redundancy) on decomposition and the temporal stability of ecosystem processes received little scientific attention. Therefore, this study aims to evaluate the effect of an increase in bacterial community resemblance on both decomposition and the stability of bacterial metabolism in aquatic sediments. To this end, we performed laboratory microcosm experiments in which we examined the influence of bacterial consortia differing in number and composition of species on bacterial activity (Electron Transport System Activity, ETSA), dissolved organic carbon production and wavelet transformed measurements of redox potential (Eh). Single substrate affinities of the individual bacterial species were determined in order to calculate the metabolic diversity of the microbial community. Results presented here indicate that bacterial activity and organic matter decomposition increase with widening of the resource niche breadth, and that metabolic stability increases with increasing overlap in bacterial resource niches, hinting that resource niche overlap can promote the stability of bacterial community metabolism.

Laboratory algal bioassays using PAM fluorometry: Effects of test conditions on the determination of herbicide and field sample toxicity

Pulse Amplitude Modulation (PAM) fluorometry, based on chlorophyll a fluorescence, is a frequently used technique in algal bioassays to assess toxicity of single compounds or complex field samples. Several test conditions can influence the test results, and because a standardized test protocol is currently lacking, linking the results of different studies is difficult. Therefore, the aim of the present study was to gain insight into the effects of test conditions of laboratory algal bioassays using PAM fluorometry on the outcome of toxicity tests. To this purpose, we described the results from several pilot studies on test development in which information is provided on the effects of the main test factors during the pretest phase, the test preparation, the exposure period, and the actual measurement. The experiments were focused on individual herbicides and complex field samples and included the effects of culturing conditions, cell density, solvent concentration, exposure time, and the presence of actinic light. Several of these test conditions were found to influence the outcome of the toxicity test, and the presented information provides important background information for the interpretation of toxicity results and describes which test conditions should be taken into account when using an algal bioassay with PAM fluorometry. Finally, the application of PAM fluorometry in algal toxicity testing is discussed.

Cell kinetics during regeneration in the sponge Halisarca caerulea: how local is the response to tissue damage?

Sponges have a remarkable capacity to rapidly regenerate in response to wound infliction. In addition, sponges rapidly renew their filter systems (choanocytes) to maintain a healthy population of cells. This study describes the cell kinetics of choanocytes in the encrusting reef sponge Halisarca caerulea during early regeneration (0–8 h) following experimental wound infliction. Subsequently, we investigated the spatial relationship between regeneration and cell proliferation over a six-day period directly adjacent to the wound, 1 cm, and 3 cm from the wound. Cell proliferation was determined by the incorporation of 5-bromo-2′-deoxyuridine (BrdU). We demonstrate that during early regeneration, the growth fraction of the choanocytes (i.e., the percentage of proliferative cells) adjacent to the wound is reduced (7.0 ± 2.5%) compared to steady-state, undamaged tissue (46.6 ± 2.6%), while the length of the cell cycle remained short (5.6 ± 3.4 h). The percentage of proliferative choanocytes increased over time in all areas and after six days of regeneration choanocyte proliferation rates were comparable to steady-state tissue. Tissue areas farther from the wound had higher rates of choanocyte proliferation than areas closer to the wound, indicating that more resources are demanded from tissue in the immediate vicinity of the wound. There was no difference in the number of proliferative mesohyl cells in regenerative sponges compared to steady-state sponges. Our data suggest that the production of collagen-rich wound tissue is a key process in tissue regeneration for H. caerulea, and helps to rapidly occupy the bare substratum exposed by the wound. Regeneration and choanocyte renewal are competing and negatively correlated life-history traits, both essential to the survival of sponges. The efficient allocation of limited resources to these life-history traits has enabled the ecological success and diversification of sponges.

Suspended particle and pathogen peak discharge buffering by a surface-flow constructed wetland

Constructed wetlands (CWs) have been shown to improve the water quality of treated wastewater. The capacity of CWs to reduce nutrients, pathogens and organic matter and restore oxygen regime under normal operating conditions cannot be extrapolated to periods of incidental peak discharges. The buffering capacity of CWs during peak discharges is potentially a key factor for water quality in the receiving waters. Therefore, the aim of the present study was to investigate the behaviour of peak discharges of suspended particles, (associated) physiochemical parameters and pathogenic organisms from a wastewater treatment plant (WWTP) in a full scale constructed wetland (CW). By mixing clarified water and sludge rich water from the settlement tank of the WWTP, the suspended particle concentration was increased for 8 h from ± 3.5 to ± 230 mg L(-1), and discharged into a full scale horizontal surface flow constructed wetland. An increase of suspended particle concentration following the peak discharge concurred with increases in turbidity and oxygen demand, total nutrient load (nitrogen, phosphorus and carbon) and pathogens (Escherichia coli and Enterococci). Temperature, pH, conductivity and dissolved nutrient concentrations (nitrogen, phosphorus and carbon) were however unaffected by the initial peak discharge. After retention in the unvegetated ponds (the first CW compartment) the applied suspended particle peak with a total load of 86.2 kg was reduced by >99%. Similar peak buffering was observed for the turbidity, oxygen demand and settable volume. Simultaneously dissolved nutrient concentrations increased, indicating partial mineralization of the suspended particles during retention in the unvegetated ponds. The peak buffering of pathogens was lower (40-84%), indicating differences in removal processes between other suspended particles and pathogens. The results indicated that the suspended particles were probably mostly removed by sedimentation and mineralization, where pathogens were more likely buffered by biofilm retainment, mortality and predation, mainly in reed ditches. After passing through the total CW the residuals of the suspended particle peak discharge were temporal increased concentrations of inorganic carbon (IC), NH(4) and E. coli (respectively 11%, 17% and 160% higher than steady state concentrations). The observations support the positive role of CWs for effective buffering of wastewater discharge peaks.