Incorporation and utilization of bacterial lipids in theSolemya velum symbiosis

We undertook a detailed analysis of the lipid composition ofSolemya velum (Say), a bivalve containing endosymbiotic chemoautotrophic bacteria, in order to determine the presence of lipid biomarkers of endosymbiont activity. The symbiont-free clamMya arenaria (L.) and the sulfur-oxidizing bacteriumThiomicrospira crunogena (Jannasch et al.) were analyzed for comparative purposes. The ¹³C ratios of the fatty acids and sterols were also measured to elucidate potential carbon sources for the lipids of each bivalve species. Both fatty acid and sterol composition differed markedly between the two bivalves. The lipids ofS. velum were characterized by large amounts of 18: 17 (cis-vaccenic acid), 16:0, and 16 : 17 fatty acids, and low concentrations of the highly unsaturated plant-derived fatty acids characteristic of most marine bivalves. Cholest-5-en-3-ol (cholesterol) accounted for greater than 95% of the sterols inS. velum. In contrast,M. arenaria had fatty acid and sterol compositions similar to typical marine bivalves and was characterized by large amounts of the highly unsaturated fatty acids 20 : 53 and 22 : 63 and a variety of plant-derived sterols. The fatty acids ofT. crunogena were similar to those ofS. velum and were dominated by 18:17, 16:0 and 16:17 fatty acids. Thecis-vaccenic acid found inS. velum is almost certainly symbiontderived and serves as a potential biomarker for symbiontlipid incorporation by the host. The high concentrations ofcis-vaccenic acid (up to 35% of the total fatty acid content) in both symbiont-containing and symbiont-free tissues ofS. velum demonstrate the importance of the endosymbionts in the lipid metabolism of this bivalve. The presence ofcis-vaccenic acid in all the major lipid classes ofS. velum demonstrates both incorporation and utilization of this compound. The ¹³C ratios of the fatty acids and sterols ofS. velum were significantly lighter (–38.4 to –45.3) than those ofM. arenaria (–23.8 to – 24.2) and were similar to the values found for the fatty acids ofT. crunogena (–45); this suggests that the lipids ofS. velum are either derived directly from the endosymbionts or are synthesized using endosymbiontderived carbon.Woods Hole Oceanographic Institution Contribution No. 7356Please address all correspondence and reprint requests to Dr Conway at her present address: Department of Biological Sciences, University of Pittsburgh, Pennsylvania 15260, USA     

Utilization of a fumigated sediment by two benthic deposit-feeders:Abra alba (Mollusca: Bivalvia) andEupolymnia nebulosa (Annelida: Polychaeta)

The present study tested the utilization of dead microbial biomass by two benthic deposit-feeders:Abra alba (Wood) (Mollusca: Bivalvia) andEupolymnia nebulosa (Montagu) (Annelida: Polychaeta). Clams were collected in the Canet lagoon during spring 1989. Worms were collected in the Port-Vendres harbour during spring 1989. The¹⁴C-labelled (glutamic acid, 24 h) sediment used during the study was sterilized with 1% chloroform, washed with sterile seawater, and dried (60°C; 48 h). This sterilisation procedure, called fumigation is the least harmful to the sediment (Novitsky 1986). Both clams and worms were incubated in the presence of the fumigated sediment for 5, 10, 20, and 50 h. At the end of each experiment we recorded the radioactivity in four compartments: (1) sediment, (2) dissolved organic matter (DOM), (3) CO₂, and (4) animals. The radioactivity of the sediment was subdivided into five fractions: (i) soluble in 2N HCl, (ii) soluble in hot 5% trichloroacetic acid (TCA), (iii) soluble in 1N NaOH, (iv) soluble in hot 6N HCl, (v) residual (after combustion in a Leco carbon analyser). In the first set of experiments, after 20 h of incubation, 5.4 and 4.7% of the total radioactivity was taken up by clams and worms, respectively. However, a model revealed that this uptake could have been correlated with the release of radiolabelled DOM (33% of total radioactivity during the first 5 h). In order to test this assumption, we used the same protocol with three additional washes of the fumigated sediment. This resulted in a significantly lower uptake by the clams (1.9% of the total radioactivity byt = 50 h), whereas the worms exhibited an uptake similar to that in the initial experiment (5.1% of total radioactivity byt = 50 h). These results underline the importance of considering interactions with DOM when applying radiotracer techniques to the study of benthic food chains. The average ingestion rates of fumigated sediment byA. alba andE. nebulosa were 5.2 10^–2 mg sediment dry wt mg^–1 clam h^–1 and 3.5 10^–2 mg sediment dry wt mg^–1 worm h^–1, respectively, which is comparable to previous data reported for other deposit-feeding bivalves and polychaetes feeding on natural sediment or detritus. The low radioactivity recorded for CO₂ together with the similarity of the changes in the partitioning of the radioactivity within the sediment between control experiments and experiments carried out in the presence of clams or worms suggest low assimilation efficiencies. Therefore, the present study supports the fact that dead microbial biomass does not constitute an important food source for benthic deposit-feeders.     

Temperature acclimation of respiration and photosynthesis in the brown algaLaminaria saccharina

Sporophytes of the brown algaLaminaria saccharina (L.) Lamour grown at 15°C contained significantly more chlorophylla (chla) than did similar plants grown at 5°C. The increase in chla in 15°C plants was due to increased numbers of photosystem II reaction centes, and possibly to increased photosynthetic unit size, compared with 5°C plants. These changes were associated with increased values (photosynthetic efficiencies) in 15°C-grownL. saccharina relative to 5°C-grown plants. The changes in together with reduced respiration rates allowed 15°C-grownL. saccharina to achieve net photosynthesis and light-saturated photosynthesis at a lower photon fluence rate (PFR) than 5°C plants when both groups were assayed at the same temperature (15°C). The photon fluence rates necessary to reach the compensation point and achieve light-saturated photosynthesis (I _c andI _k , respectively) increased with increasing incubation temperature inL. saccharina grown at both 5 and 15°C. However, acclimation responses to growth temperature compensated for the short-term effect of temperature onI _c andI _k . Consequently, plants grown at 5 and 15°C were able to achieve similar rates of light-limited photosynthesis, and similarI _c andI _k values at their respective growth temperatures. These responses are undoubtedly important for perennial seaweeds such asL. saccharina, which frequently grow in light-limited habitats and experience pronounced seasonal changes in water temperature.Please address all correspondence and requests for reprints to I.R. Davison     

Effects of Sphagnum moss and urban runoff on bioavailability of lead and zinc from acidic wetlands of the New Jersey pinelands

The effects of Sphagnum and urban runoff on the bioavailability of metals were tested by adding PbCl2 and ZnCl2 to laboratory microcosms constructed of peat substrate with or without live Sphagnum spp. and planted with Acer rubrum L. seedlings or Vaccinium macrocarpon Ait. shoots. We hypothesized that Sphagnum would increase bioavailability through its acidifying action, while urban runoff would decrease availability through increases in alkalinity. Metals were more available to the test plants treated with acidic swampwater than with alkaline runoff. Sphagnum moss caused increases in tissue concentrations in V. macrocarpon, but not in A. rubrum. The latter species took up more metals when grown on substrate from sites receiving runoff, whereas the former took up more metals from substrate from undisturbed sites, despite the lower substrate metal concentrations. Differences in uptake by the two species may reflect differences in their ability to root in the Sphagnum mat. The results demonstrate that Sphagnum spp. exerts strong species-specific effects on the uptake of metals by vascular plants, and that plant species native to acidic wetlands vary widely in their response to metals in the substrate.     

Effects of elevated CO2 concentration on the polyamine levels of field-grown soybean at three O3 regimes

Effects of increased ozone (O3) and carbon dioxide (CO2) on polyamine levels were determined in soybean (Glycine max L. Merr. cv. Clark) grown in open-top field chambers. The chamber treatments consisted of three O3 regimes equal to charcoal filtered (CF), non-filtered (NF), and non-filtered plus 40 nl litre(-1) O3 and CO2 treatments equal to 350, 400 and 500 microl litre(-1) for a total of nine treatments. Leaf samples were taken at three different times during the growing season. Examination of growth and physiological characteristics, such as photosynthesis, stomatal resistance, and shoot weight, revealed that increasing CO2 ameliorated the deleterious effects of increased O3. Results from the initial harvest, at the pre-flowering growth stage (23 days of treatment), showed that increasing O3 at ambient CO2 caused increases in putrescine (Put) and spermidine (Spd) of up to six-fold. These effects were lessened with increased CO2. Elevated CO2 increased polyamines in plants treated with CF air, but had no effect in the presence of ambient or enhanced O3 levels. Leaves harvested during peak flowering (37 days of treatment) showed O3-induced increases in Put and Spd at ambient CO2 concentrations. However, increased CO2 levels inhibited this response by blocking the O3-induced polyamine increase. Leaves harvested during the pod fill stage (57 days of treatment) showed no significant O3 or CO2 effects on polyamine levels. Our results demonstrate that current ambient O3 levels induce the accumulation of Put and Spd early in the growing season and that further increases in O3 could result in even greater polyamine increases. These results are consistent with a possible antiozonant function for polyamines. The ability of increased CO2 to protect soybeans from O3 damage, however, does not appear to involve polyamine accumulation.     

Effect of mecoprop drift on some plant species of conservation interest when grown in standardized mixtures in microcosms

There has been an increasing awareness of potential impacts of herbicide drift on to vegetation in nature reserves and field margin habitats adjacent to treated areas. Previous work using single species bioassays has suggested that the effects of a single drift event are confined close to the sprayer (< 10 m). In the present study eight native dicotyledonous species, with and without a perennial grass (Lolium perenne), were grown in standardized microcosms in order to study (1) the effects of herbicide drift where plants were exposed to competition, and (2) the effects of a second spray application. The microcosms were arranged downwind (0-8 m) of a standard agricultural sprayer applying mecoprop at recommended rates. The effects of the herbicide drift on foliar symptoms of plant damage and end-of-season yield were assessed in each of two years. The main conclusions were that (1) growth of Stachys sylvatica and Lolium perenne (where sown) was enhanced near the sprayer and, (2) six other species (Digitalis purpurea, Galium mollugo, Hypericum hirsutum, Lychnis flos-cuculi, Primula veris and Ranunculus acris) showed some evidence of reduction in either performance (assessed non-destructively after a single exposure) or yield after two exposures. Three species (Lychnis flos-cuculi, Primula veris and Ranunculus acris) showed a reduction in flowering performance. Thus, mecoprop drift affected the aesthetic appearance, possibly the fecundity of some species and the balance between species in these microcosm experiments. The implications of these results for the persistence of attractive plant communities in sensitive areas are discussed.     

Development of monitoring technology for airborne particulate matter

Particulate matter suspended in the air has adverse effects onhuman health. Its level of concentration is an important parameter in evaluating the degree of hazard it poses to the atmosphere. Conventional methods used in measuring particulatematter are often filter-based, which indicates some disadvantagesbecause such a base requires labor and time. In this study, to achieve real-time measurements, a new electrical method was developed for measuring PM10 and PM2.5 concentrations. The basicprinciple is to electrically charge particles passing through thePM inlet using a corona charger and measure the currents createdby charged particles to obtain the number concentration of particulate matter. A new type inlet based on the particle cupimpactor configuration was designed and its performance was evaluated. A unipolar diffusion charger was developed and thecharger's efficiency was determined experimentally in terms ofPn, which represents the penetration through the charger,P, times the average charge number acquired by a particle,n, for different particle sizes. The correlation was constructed between the PM10 (or the PM2.5) mass concentrationsand the electrical currents due to particles, which were chargedby the diffusion charger.     

Recycling of petroleum-contaminated sand

The environmental impact of using petroleum-contaminated sand (PCS) as a substitute in asphalt paving mixtures was examined. An appreciable component of PCS is oily sludge, which is found as the dregs in oil storage tanks and is also produced as a result of oil spills on clean sand. The current method for the disposal of oily sludge is land farming. However, this method has not been successful as an oil content of < 1% w/w is required, and difficulty was encountered in reaching this target. The reuse of the sludge in asphalt paving mixtures was therefore considered as an alternative. Standard tests and environmental studies were conducted to establish the integrity of the materials containing the recycled sludge. These included physical and chemical characterization of the sludge itself, and an assessment of the mechanical properties of materials containing 0%, 5%, 22% and 50% oily sludge. The blended mixtures were subjected to special tests, such as Marshall testing and the determination of stability and flow properties. The experimental results indicated that mixtures containing up to 22% oily sludge could meet the necessary criteria for a specific asphalt concrete wearing course or bituminous base course. To maximize the assay from the recycled material, the environmental assessment was restricted to the 50% oily sludge mixture. Leachates associated with this particular mixture were assayed for total organic residue and certain hazardous metal contaminants. The results revealed that the organics were negligible, and the concentrations of the metals were not significant. Thus, no adverse environmental impact should be anticipated from the use of the recycled product. Our research showed that the disposal of oily sludge in asphalt paving mixtures could possibly yield considerable savings per tonne of asphalt concrete, and concurrently minimize any direct impact on the environment.