Quantitative PCR to estimate copepod feeding

Copepods play a central role in marine food webs as grazers of plankton and as key prey for many predators. Therefore, quantifying their specific trophic interactions is critical for understanding the role of copepods in ocean processes. However, because of methodological constraints, it remains difficult to investigate in situ copepod feeding without reliance on laborious intrusive and potentially biased incubation approaches. Recent advances in PCR-based methodologies have demonstrated the feasibility of directly identifying copepod diets based on prey DNA sequences. Yet, obtaining quantitative information from these approaches remains challenging. This study presents results of systematic efforts to develop a quantitative PCR (qPCR) assay targeted to 18S rRNA gene fragments to estimate copepod gut content of specific species of prey algae. These results were first compared to gut content estimates based on fluorescence in the copepod Calanus finmarchicus fed monocultures of two different microalgae species in controlled laboratory studies. In subsequent field studies, we compared feeding rates obtained by microscopy and qPCR for Temora longicornis and Acartia clausi feeding on the haptophyte Phaeocystis globosa in natural blooms. These investigations demonstrate a semi-quantitative relationship between gut content estimates derived from qPCR, gut pigment, and direct microscopy. However, absolute estimates of gut content based on qPCR methodology were consistently lower than expected. This did not appear to be explained by the extraction methods used, or interference by non-target (predator) DNA in the PCR reactions, instead suggesting digestion of prey-specific nucleic acids. Furthermore, the 18S rDNA target gene copy number of the phytoplankton varied with growth phase. Nonetheless, when prey target gene copy number in the ambient water is quantified, the qPCR-approach can be compared to other methods, and then used to semi-quantitatively estimate relative copepod grazing on specific prey in situ without involving further incubations. A distinct advantage of a DNA-based molecular approach compared to gut fluorescence and direct microscopic observation, is the ability to detect non-pigmented and macerated prey. Future studies should aim to correct for breakdown in prey DNA and perform extensive calibrations to other methods in order to achieve a quantitative measure of feeding rates in situ.     

Seasonal evolution of gas-phase PCB concentrations in the Venice Lagoon area

The concentrations of gas-phase PCBs were measured from March 2002 to June 2003 at two sites of the Venice Lagoon and at one site of the Euganei hills. The aims of this study were to evaluate the various gas-phase PCB sources, the spatial and temporal variability of PCB concentrations in the gas-phase that enter the Venice Lagoon atmosphere and the influence of the air temperature on PCB trends. The highest annual average concentration of summation PCBs was observed at the station directly influenced by "urban" sources with values about 3 and 1.5 times higher compared to the concentrations found at the stations where "marine" sources and "not subjected to direct local sources" were respectively sampled from. The temporal trends of summation PCBs concentrations were similar at the three sampling stations corresponding to seasonal temperature changes. Greater concentrations occurred in the summer and first autumn months while the lower ones occurred in late autumn and winter. The temperature dependences were investigated using plots of the natural logarithm of the summation PCBs concentrations vs. reciprocal mean temperatures.     

Isomeric analysis of BTEXs in the atmosphere using beta-cyclodextrin capillary chromatography coupled with thermal desorption and mass spectrometry

An analytical method capable of determining trace levels of BTEX-aromatics (benzene, toluene, ethylbenzene, m-, p- and o-xylenes) in the atmosphere with as high resolution as possible has been developed. The method is based on the preconcentration of air samples using a multibed tube (Carbopack C, Carbograph 1) at ambient temperature, followed by thermal desorption, and analysis of aromatic species by a beta-cyclodextrin capillary chromatography coupled with mass spectrometry. The resolution achieved was sufficient for individual separation of BTEXs as well as m- and p-xylenes. The BTEX-ratios have been determined in an air tunnel and in on-road, suburban and rural forest atmosphere. The ethylbenzene/m-xylene ratios could provide a deep insight into anthropogenic related NMHC patterns at different locations and under different meteorological conditions and may reflect photochemical processes in the best way.     

Relevance of wound-activated compounds produced by diatoms as toxins and infochemicals for benthic invertebrates

Plants evolve the production of wound-activated compounds (WACs) to reduce grazing pressure. In addition, several plant-produced WACs are recognized by various invertebrates, playing the role of infochemicals. Due to co-evolutionary processes, some invertebrates recognize plant infochemicals and use them to identify possible prey, detect the presence of predators or identify algae containing various classes of toxic metabolites. Different metabolites present in the same algae can play the role of toxins, infochemicals or both simultaneously. We investigated the infochemical activity of compounds extracted from three diatoms epiphytes of the seagrass Posidonia oceanica, by conducting choice experiments on invertebrates living in the same community or in close proximity. Furthermore, the specific toxicity of the extracts obtained from the same algae was tested on sea urchin embryos using a standard bioassay procedure, to detect the presence of toxins. The comparison of the two effects demonstrated that invertebrates are subjected to diatom wound-activated toxicants when these algae are not associated with their own habitat, but they are able to recognize volatile infochemicals derived from diatoms associated with their habitats. The specific toxicity of WACs was shown to be inversely correlated to the perceptive ability of invertebrates towards volatile compounds liberated by the same algae. Hence, when the recognition of specific algae by a given invertebrate species evolves, their detrimental effects on the receiving organism may be lost.     

Biological benthic tools as indicators of coastal marine ecosystems health

Coastal marine ecosystems are increasingly subjected to environmental stress and degradation due to pollution. Several research programmes have addressed this problem and produced relevant data sets for specific areas, often including consistent sets of environmental and biological variables. The value of existing information gathered from these types of data can be largely increased by combining them into a common data set to determine globally applicable relationships. To perform this exercise, the Intergovernmental Oceanographic Commission (IOC) of UNESCO has recently formed the Ad hoc Study Group on Benthic Indicators (http://www.ioc.unesco.org/benthicindicators) with the aim of developing robust indicators of benthic health. In this paper, initial products and ongoing activities of this international initiative are described and discussed. An expansion of initial IOC/UNESCO research on benthic fauna-organic carbon relationships is also presented. As part of this follow-up research, the relationship between total organic carbon concentrations of sediment and abundance, biomass and species diversity of benthic macrofauna was evaluated using data sets from 2 different regions of the world comprising 3 different coastal marine environments. The ability of identifying threshold levels in selected variables that could serve as indicators of related adverse environmental conditions leading to stress in the benthos is envisaged within the frame of a larger joint analysis, carried out by the IOC/UNESCO Study Group on Benthic Indicators, of merged data sets from several coastal regions worldwide.     

High rate mesophilic, thermophilic, and temperature phased anaerobic digestion of waste activated sludge: a pilot scale study

The paper reports the findings of a two-year pilot scale experimental trial for the mesophilic (35°C), thermophilic (55°C) and temperature phased (65+55°C) anaerobic digestion of waste activated sludge. During the mesophilic and thermophilic runs, the reactor operated at an organic loading rate of 2.2 kgVS/m(3)d and a hydraulic retention time of 20 days. In the temperature phased run, the first reactor operated at an organic loading rate of 15 kgVS/m(3)d and a hydraulic retention time of 2 days while the second reactor operated at an organic loading rate of 2.2 kgVS/m(3)d and a hydraulic retention time of 18 days (20 days for the whole temperature phased system). The performance of the reactor improved with increases in temperature. The COD removal increased from 35% in mesophilic conditions, to 45% in thermophilic conditions, and 55% in the two stage temperature phased system. As a consequence, the specific biogas production increased from 0.33 to 0.45 and to 0.49 m(3)/kgVS(fed) at 35, 55, and 65+55°C, respectively. The extreme thermophilic reactor working at 65°C showed a high hydrolytic capability and a specific yield of 0.33 g COD (soluble) per gVS(fed). The effluent of the extreme thermophilic reactor showed an average concentration of soluble COD and volatile fatty acids of 20 and 9 g/l, respectively. Acetic and propionic acids were the main compounds found in the acids mixture. Because of the improved digestion efficiency, organic nitrogen and phosphorus were solubilised in the bulk. Their concentration, however, did not increase as expected because of the formation of salts of hydroxyapatite and struvite inside the reactor.     

Food resource partitioning in a Mediterranean demersal fish assemblage: the effect of body size and niche width

We investigated the effects of body size, feeding strategy and depth distribution on the trophic resource partitioning among the 26 dominant fish consumers in a fish assemblage on the central Mediterranean shelf-break. The fish assemblage was structured in two major trophic guilds: epibenthic and zooplanktonic feeders, according to the position of each predator along the benthos–plankton gradient. Within each main guild, the species were segregated along a prey-size or fish-size gradient into five further guilds. Fish size and prey size were strongly correlated, indicating that the prey-size niche can be well explained by predator size. Fish consumers showed a significant negative correlation between the similarity in prey type and the similarity in depth distribution; most species with similar trophic preferences segregated along the depth dimension. The only predators overlapping in both food and depth preferences were those with a more specialist trophic behavior. These results suggest that fish body size and depth preferences are the two main niche dimensions, explaining a large part of the coexistence between the Mediterranean shelf-break fish consumers.     

Response surface models for the formation of PCDD and PCDF in a pilot plant combustion of MSW

Experimental data on PCDD and PCDF emissions in raw gases from municipal solid waste combustion in a pilot plant were processed by Response Surface models to gain information on process parameter values which minimize both the total PCDD/PCDF concentrations and those of a selected congener group.     

Environmental Pollution Due to Natural Factors: A Case Study in A Volcanic Area (Vulcano Island, Italy)

A major influence on the environment of active volcanic areas is expected from continuous input of chemical species pertaining to fumarolic activities occurring during inter-eruptive intervals.

The systematic investigations carried out during thirteen years in the island of Vulcano (Italy) showed also substantial contributions to phreatic waters and soils of minor constituents and trace metals from volcanic rocks with no negligible influence on growing crops.

Greater extents of these phenomena have been observed for products of hydromagmatic volcanic activity or of long-lasting weathering processes.     

Visible light driven mineralization of spiramycin over photostructured N-doped TiO2 on up conversion phosphors

A novel visible light-active photocatalyst formulation(NdT/OP) was obtained by supporting N-doped TiO_2(NdT) particles on up-conversion luminescent organic phosphors(OP). The photocatalytic activity of such catalysts was evaluated for the mineralization process of spiramycin in aqueous solution. The effect of NdT loading in the range 15–60 wt.% on bulk and surface characteristics of NdT/OP catalysts was investigated by several chemicophysical characterization techniques. The photocatalytic performance of NdT/OP catalysts in the removal of spyramicin from aqueous solution was assessed through photocatalytic tests under visible light irradiation. Total organic carbon(TOC) of aqueous solution,and CO and CO_2 gas concentrations evolved during the photodegradation were analyzed. A dramatic enhancement of photocatalytic activity of the photostructured visible active NdT/OP catalysts,compared to NdT catalyst,was observed. Only CO_2 was detected in gas-phase during visible light irradiation,proving that the photocatalytic process is effective in the mineralization of spiramycin,reaching very high values of TOC removal. The photocatalyst NdT/OP at 30 wt.% of NdT loading showed the highest photocatalytic activity(58%of TOC removed after 180 min irradiation against only 31% removal after 300 min of irradiation of NdT). We attribute this enhanced activity to the high effectiveness in the utilization of visible light through improved light harvesting and exploiting. OP particles act as "photoactive support",able to be excited by the external visible light irradiation,and reissue luminescence of wavelength suitable to promote NdT photomineralization activity.