β-glucanase activity in the seastar Pisaster ochraceus

Specimens of Pisaster ochraceus, an intertidal carnivorous seastar, were collected in northern Puget Sound at Post Point, near Bellingham, Washington, USA, in November 1993, and used in experimentation through September 1994. Ammonium sulfate fractions (60% precipitates) from excised pyloric caeca (digestive glands) possessed -glucanase activity directed toward such -linked substrates as carboxymethylcellulose (CMC), partially-digested cellulose (cellodextrins) and the synthetic substrate p-nitrophenyl--D-glucoside (PNDG). pH-activity and pH-stability maxima were at pH 6 and 5, respectively. No hydrolytic activity was detectable on native cellulose, native chitin or on cationic-modified celluloses such as DEAE-cellulose. Gel filtration (Sephadex G-100) of this 60% precipitate followed by DEAE-Sephadex column chromatography revealed heterogeneity of -glucanase activity, with an indication of up to 11 distinct fractions with variable activity directed toward the substrates indicated above (CMC, cellodextrins and PNDG). These fractions displayed differing ratios of hydrolytic activity on these substrates. None of the isolated fractions displayed activity toward either native cellulose or native chitin. Gut extracts from the edible mussel (Mytilus edulis), a common prey species of P. ochraceus, were shown to possess partially digested cellulose (cellodextrins). These cellodextrins were further degraded by seastar -glucanases in vitro. The digestion of the gut contents of ingested prey provides a possible adaptive explanation for the presence of -glucanase activity in a carnivore.     

Laboratory evidence of natural remobilization of multicomponent DNAPL pools due to dissolution

Mixtures of dense non-aqueous phase liquids (DNAPLs) trapped in the subsurface can act as long-term sources of contamination by dissolving into flowing groundwater. In general, the components of higher solubility are removed more quickly, thus altering the composition of the remaining DNAPL, and possibly leading to changes in its physical properties. Through the development of a simple compositional model, Roy et al. [J. Contam. Hydrol. 2002 (59) 163] showed that preferential dissolution of a mixed DNAPL could potentially result in changes in density and interfacial tension that could subsequently lead to remobilization of an initially static DNAPL pool. The laboratory experiments presented in this next paper provide a proof-of-concept for the previously presented theory, demonstrating and quantifying this process of remobilization. In addition, the experiments provide a data set for evaluation of the model presented by Roy et al. [J. Contam. Hydrol. 2002 (59) 163]. In the four experiments, a DNAPL pool comprised of tetrachloroethene and benzene was created as an open pool overlying glass beads within a water-saturated 2-D flow box. Experiments included rectangular and triangular pools. In each of the experiments, remobilization (as breakthrough) was observed more than 2 weeks after formation of the initial pool. During each experiment, the pool height declined as mass was lost by dissolution, while sampling indicated a decrease in the mole fraction of benzene, the more soluble component. Small protuberances formed along the bottom of the pool as its composition changed with time and the displacement pressure was achieved for various pore throats. Eventually one of the protuberances extended further, forming a finger (breakthrough). In general, the pool emptied as the finger proceeded further into the beads. It was also shown theoretically and experimentally that remobilization will occur sooner for pools with a triangular (pointing down), rather than rectangular, shape. The experimental results were simulated using the model developed by Roy et al. [J. Contam. Hydrol. 2002 (59) 163]. The model matched the observations well, suggesting that it accurately represents the primary mechanisms involved with natural remobilization under the conditions of the study.     

The use of sediment analogues to study the uptake of pollutants by chironomid larvae

A technique is described that uses artificial resin beads with known surface properties to investigate the factors influencing the bioaccumulation of pollutants from sediments. One advantage of this technique is that it provides a standard procedure against which it is possible to calibrate natural sediments with their diverse properties. The method has been used on third instar larvae of the midge Chironomus riparius and the results are compared with previous studies on the worm Lumbriculus variegatus. The use of a standard test using resin beads as a substitute for natural sediment allows comparisons to be made between species and substrates. Thus, the bioaccumulation factors for the midge larvae are much smaller than those of the worm and this correlates with the ability of the insect larva to detoxify many pollutants. It is also possible to use the test to identify if ingestion of the sediment increases the bioaccumulation of contaminants and whether this involves the release of pollutants by digestive processes or not.     

Nitrogen uptake and growth of the germlings and mature thalli of Fucus distichus

Fucus distichus L. was collected near Vancouver, Canada, in late fall and early winter, 1981. The effects of the forms of nitrogen (nitrate, ammonium or urea) and periodic exposure to air on growth, rhizoid development and nitrogen uptake in germlings was investigated. Gamete release, fertilization, germination and germling growth had no requirement for a specific form of nitrogen. Periodic exposure to air increased secondary rhizoid development twofold. Nitrate and ammonium uptake rates of the germlings were higher than for the mature thalli (20 to 40 times for nitrate and 8 times for ammonium), while the halfsaturation constant (K _s) values for nitrate were similar (1 to 5 M). The germlings showed saturable uptake kinetics but the mature thalli did not. When germlings were exposed to air it caused a 70% decrease in nitrate uptake, but not change in ammonium uptake. Ammonium uptake in the mature thalli was proportional to the ambient ammonium concentration. Nitrate uptake in the mature thalli appeared to follow saturation kinetics at low nitrate concentrations, but showed a non-saturable component at concentrations greater than 10 M. Presence of ammonium inhibited nitrate uptake by the mature plants but not by the germlings.     

Demonstration,using Desulfobacter sp., of two pools of acetate with different biological availabilities in marine pore water

Desulfobacter sp. (Strain 3ac10), an acetate-utilizing sulphatereducing bacteria, was added to sterile marine pore water spiked with ¹⁴C-acetate, and changes in both the natural acetate pool and the added ¹⁴C-acetate were measured over time. Initially, both the added ¹⁴C-acetate and the chemically measured acetate were rapidly mineralized, but then the rate of removal decreased and a significant amount (approximately 20%) of both ¹⁴C-acetate and chemically measured acetate remained unmetabolised. In a replicate experiment, approximately 50% of the acetate was not metabolised. Kinetic analysis of the data indicated that there were two pools of acetate in the original pore water, a biologically available pool (which is rapidly metabolised) and a recalcitrant pool (which is only very slowly metabolised). Addition of ¹⁴C-acetate after the biologically available acetate had been removed resulted in rapid removal of the added acetate but no change in the recalcitrant acetate pool. The implications of this data to radiolabelled techniques of measuring in situ acetate turnover are discussed.     

A ciliate red tide at Barrow,Alaska

In September 1968 the first occurrence of extensive red water in the Arctic Ocean in the vicinity of Point Barrow, Alaska, was recorded. The organism causing this water discoloration was a fairly large (100 to 150 ) ciliated protozoan, with chlorophyll-containing endosymbionts. This ciliate, which is not identical to the commonly reported Mesodinium or Cyclotrichium species, is described in this paper, but cannot be identified with any organism described in the literature. Concentrations of dissolved inorganic nutrients and trace metals were determined on water samples obtained from the red water and also in the clear water adjacent to it. These data are discussed relative to the hydrographic conditions in the Chuckchi Sea around Point Barrow. A direct species' analysis by microscopic methods indicated that the plankton were quite similar in all the water samples except for the aforementioned red ciliate, which accounted for over 90% of the total cellular organic carbon in the samples from the red water. Chemical analyses indicated that the red tide ciliate contained approximately 51% protein, 33% lipid, 8% carbohydrate, 0.2% chlorophyll a, and 1.2% DNA.