Physiological background for using freshwater mussels in monitoring copper and lead pollution

In studying the effect of copper (10 ± 0.57 µg Cu l^–1 and 100 ± 3.01 µg Cu l^–1) and lead (50 ± 1.12 µg Pb l^–1 and 500 ± 12.5 µg Pb l^–1) on the filtration activity of Anodonta cygnea L. it was found that both heavy metals resulted in significant shortening of the active periods, but little change occurred in the length of the rest periods. The concentrations of copper and lead were measured in the gill, foot, mantle, adductor muscle and kidney for 840 hours of exposure to 10.9 ± 5 µg Cu l^–1 and 57.0 ± 19 µg Pb l^–1 as well as during subsequent depuration. Uptake was observed after 72 hours of exposure. The highest copper concentration (59.1 ± 16.2 µg Cu g^–1) was measured at 672 h in the mantle, and the highest lead value (143 ± 26.1 µg Pb^–1) was obtained in the kidney. Depuration of copper was fastest from the foot, and from the adductor muscle for lead. The gill had the longest half-depuration time (> 840 h for copper and > 672 h for lead).     

Effects of diet and heavy metals on growth rate and fertility in the deposit-feeding snail Potamopyrgus jenkinsi (Smith) (Gastropoda: Hydrobiidae)

Evidence for the influence of food type and heavy metals on shell growth and fertility is presented for a freshwater population of the snail P. jenkinsi. When fed an excess of lettuce or lamb heart (protein source), growth rates were higher for lettuce. Highest growth rates occurred at a diet of lettuce plus lamb heart. Fertility was favoured by a diet of lamb heart. When fed an excess of lettuce, the EC₅₀ growth values were 16 µg Cd l^–1, 13 µg Cu l^–1, and 103 µg Zn l^–1 in lake water; snail fertility was inhibited at 25 µg Cd l^–1 and 30 µg Cu l^–1. A diet of lake detritus spiked with Cd or Cu resulted in a decrease of approximately 50% in growth rates, when compared with growth on non-spiked detritus. Spiked detritus lost metals into lake water. Food type positively interacted with metal stress, both for growth rate and fertility. The assessment of inhibitory effects of detritus contaminated either in the field or, notably, by spiking, and serving as food source for deposit feeders is hampered by sampling problems in the field and by redistribution processes of pollutants between particles and water in laboratory-scale experiments.     

Aspects toxiques du cuivre sur la biomasse et la productivité du phytoplancton de la rivière du Saguenay,Québec

In 1979 and 1980, batch culture experiments were conducted to observe the inhibitory effect of copper ion (concentrations of 10, 50, 100, 200 and 400 µg Cu · l^–1) on the standing crops and photosynthesis of phytoplankton of the Saguenay River (for 124 hours) and in Chlorella vulgaris (for 8 days). These algal assays were carried out using the surface water of the Saguenay River. In natural populatoins of phytoplankton, it was found that photosynthesis was more sensitive than growth: at the lowest concentrations, such as 10 µg Cu · 1^–1, copper seemed to increase the chlorophyll concentrations whereas the rates of primary production show a decrease of 60% with respect to the control. At higher concentrations of copper, the effect is weak in chlorophyll concentrations and more pronounced in the rates of primary production (decrease of 86 to 90%). The pennate diatoms are dominant (in all the samples) and these organisms are known as relatively resistant to copper. In Chlorella vulgaris, it was observed that with 100 µg Cu · 1^–1, chlorophyll concentrations and rates of photosynthesis respectively decrease by 63 and 99% with respect to the control. At higher concentrations of copper, a maximum decrease of 70% and 99% respectively for chlorophyll concentrations and rates of primaryproduction are observed.

     

Heterotrophic glucose uptake and respiration in Lake Kinneret

The turnover times of glucose, averaged for 0–10 m in the upper waters of Lake Kinneret and measured by the addition of single or multiple concentrations of substrate, ranged from 23 to 188 hours and 1 to 87 hours respectively. Potential uptake rates (estimated as V_max) ranged from 0.095 to 1.94 µg glucose l^–1h^–1, while measured uptake rates varied from 0.09 to 1.1 µg glucose l^–1h^–1. Concentrations of dissolved carbohydrates and glucose averaged 0.71 mg glucose equivalents l^–1 and 39 µg glucose l^–1 respectively. No evident relationships between glucose cycling and any fractions of dissolved organic matter, phytoplankton biomass or primary productivity were found. Turnover times were generally most rapid immediately after the decline of the spring Peridinium bloom. The respiration percentage of incorporated glucose ranged from 25% to 61% with highest values during the summer months. Respiration may be influenced by the nature of the indigenous bacterial population as well as by temperature. Daily heterotrophic glucose carbon uptake was about 9% of the photosynthetic incorporation and could provide a bacterial yield of about 7 × 10⁴ ml^–1d^–1.     

Batch xylitol production from wheat straw hemicellulosic hydrolysate using Candida guilliermondii in a stirred tank reactor

Batch production of xylitol from the hydrolysate of wheat straw hemicellulose using Candida guilliermondii was carried out in a stirred tank reactor (agitation speed of 300 rpm, aeration rate of 0.6 vvm and initial cell concentration of 0.5 g l^–1). After 54 h, xylitol production from 30.5 g xylose l^–1 reached 27.5 g l^–1, resulting in a xylose-to-xylitol bioconversion yield of 0.9 g g^–1 and a productivity of 0.5 g l^–1 h^–1.     

A laboratory study of feeding and assimilation in Euchlanis dilatata lucksiana

Feeding of Euchlanis dilatata lucksiana, a brachionid rotifer isolated from the Loosdrecht Lakes (The Netherlands), was examined in the laboratory using ¹⁴C-labelled food. The gut-filling time at a food concentration of 9.6 µg C ml^–1 was about 15 minutes. Animals which were fed on 3 size fractions of the lake seston (< 7 µm, 7–15 µm, and 15–33 µm) showed a clear preference (Jacobs' selectivity index) for the largest size fraction. This fraction was composed predominantly of filamentous cyanobacteria. For animals weighing 0.37–0.49 µm C ind.^–1 the daily ration (daily food consumption per unit body weight × 100) ranged from 50 to 100% at food levels of 2 mg C l^–1 and below, but increased to 150–250% at food concentrations of 5 mg C l^–1 and above. The assimilation efficiency was 100% up to 5 mg C l^–1 of food, but decreased to about 80% at higher food levels.     

Effects of a plume front on the distribution of inorganic and organic matter off the Rhône River

Hydrological and chemical structures off the Rhône River estuary resulting from the introduction of the river flow into the Mediterranean Sea are described. The effect of the fresh-water/sea-water interface on the distribution of inorganic and organic matter off the Rhône river is investigated. Strong vertical gradients of inorganic and dissolved organic matter such as lipids characterized the first few meters in this area (from 83.7 to 0.6 N-NO₃ µgat l^–1, from 6.39 to 0.92 N-NH₄ µgat l^–1 and from 299 to 73 µg l^–1 of total dissolved lipids). At the interface, substantial increases of particulate organic (PON: from 45 µg l^–1 at surface to 118 µg l^–1 at the interface, POC: from 462 to 876 µg l^–1, total particulate lipids: from 33 to 648 µg l^–1) and suspended matter in general (from 18 to 22.2 mg l^–1) were observed. High phytoplanktonic production may account for some of this enrichment, although passive accumulation might also be involved.     

Effects of copper sulphate on Tisbe holothuriae Humes (Copepoda) and development of tolerance to copper

A study was made of the toxicity of copper to the marine copepod Tisbe holothuriae Humes and the development of increased tolerance due to acclimation. In sublethal concentrations, copper causes a prolongation of maturation time and a reduction in offspring production. These effects increased proportionally to the copper concentration used, while the respiratory rate (measured at the highest concentration used: 0.008 mg l^–1 Cu) increased. A tendency for higher tolerance due to acclimation was also observed. The copper induced delay in maturation time observed in the F₂ generation, became less pronounced from F₃ onwards. In animals exposed at 0.008 mg l^–1 Cu the maturation time of F₃ was not statistically different from that of untreated animals. The LC50 48 h also presented an increase due to acclimation in animals exposed at 0.004 (F₅) and 0.008 mg l^–1 Cu (F₃ and F₅ generations), but this increase was not statistically significant.     

A comparison of the responses of two microcosm designs to a toxic input of copper

Two microcosm designs were compared for their sensitivity to toxic concentrations of copper. One design simulated a littoral zone, including macrophytes, sediment, and associated organisms. The other design used a periphyton community collected on polyurethane foam artificial substrata. Microcosms were dosed with copper sulfate (0–300 µg Cu 1^–1, nominal concentrations) and monitored for changes in several structural and process variables. Coefficients of variation of responses measured from the littoral microcosms were greater than from responses measured from the artificial-substrata microcosms. Effects were detected more frequently at lower concentrations of copper in the artificial-substrata microcosms than in the littoral microcosms. Lowest observable effect concentrations (LOECs) for measures of community structure ranged from 20.2–42.8 µg Cu 1^–1 in the artificial-substrata microcosms and from 24.0–98.5 µg Cu 1^–1 in the littoral microcosms. LOECs for measures of community process ranged from 42.8–310.3 µg 1^–1 in the artificial substrata microcosms. Significant differences from controls for community process were detected only at 304.7 µg Cu 1^–1 in the littoral microcosms. While there were differences between the two microcosm designs in the concentrations of copper that resulted in adverse effects, response trends were similar. Often, dose-response relationships between variables and copper concentrations were not log-linear, but showed stimulations at intermediate concentrations of copper (10–100 µg 1^–1, nominal concentrations). The choice of microcosm design should be dependent on the particular research question, as the designs differ in complexity and in the ease of construction and maintenance.     

On the dynamics of exploited populations ofTisbe holothuriae (Copepoda,Harpacticoidae)

A total of 90, weekly exploited populations of the harpacticoid copepodTisbe holothuriae Humes were exposed to 148, 222, 333, 500, 750 or 1125µg Cd⁺⁺ l^–1, combined with exploitation rates of 10, 30, 50, 70 or 90% under conditions of surplus food supply at 22° C and 30 S. All populations exposed to concentrations down to 500µg Cd⁺⁺ l^–1 and 3 populations (out of 15) exposed to 333µg l^–1 became extinct within the experimental period of 30 weeks. Survival time significantly depended on concentration. A recovery phase from an initially high mortality preceded eventual population extinction after adding 500µg Cd⁺⁺ l^–1. In the initial phase, higher nauplii mortality prevailed. During these experiments on acute intoxication, no relationship could be established between survival and exploitation rate. However, experiments on the effects of stepwise increases in Cd concentration (results not yet published) produced evidence of such a relationship. In spite of increased mortality, no significant numbers of dead copepods were detected in weekly samples because of their rapid decomposition and cannibalism, which depends on the amount of food available. Sampling regimes of 5 times per week yielded significant numbers of dead individuals.     

Poly(3-hydroxybutyrate) synthesis in fed-batch culture of Ralstonia eutropha with phosphate limitation under different glucose concentrations

Poly(3-hydroxybutyrate) (PHB) was produced by fed-batch cultures of Ralstonia eutropha with phosphate limitation under different glucose concentrations. When glucose was kept at 2.5 g l^–1, cell growth and PHB synthesis were limited due to the shortage of carbon source but a higher PHB content occurred in the cell-growth stage. This shows that a low glucose concentration is favorable for PHB accumulation in R. eutropha. PHB obtained with glucose at 9 g l^–1 is 1.6 times that obtained with 40 g l^–1. When glucose was in the range of 9 to 40 g l^–1, PHB concentration and productivity decreased significantly with the increase of glucose concentration. The highest PHB productivity was obtained with glucose at 9 g l^–1.     

Total organochlorine and organobromine in Finnish water courses

Methods were developed for analyzing total organochlorine (TOCl) and total organobromine (TOBr) in different kinds of water, sediment and biological samples. Sediment samples were extracted with alkaline propanone and biological samples with a neutral propanone + water solution. Neutron activation analysis (NAA) was used for the halogen determination. The detection limits for TOCl and TOBr are respectively 5 and 0.5 µg l^–1 for water (sample size 100 ml) and 0.3 and 0.1 µg g^–1 dw (sample size 1 g) for sediment and biological samples. Recoveries for common organochlorine compounds were usually over 90%.TOCl concentrations measured in waters in a natural condition were below 15 µg l^–1; and in polluted waters, from 100 to 500 µg l^–1, and occasionally to 2000 µg l^–1. TOBr concentrations were generally well below 10 µg 1^–1.     

Selective production of bikaverin in a fluidized bioreactor with immobilized Gibberella fujikuroi

The best culture medium composition for the production of bikaverin by Gibberella fujikuroi in shake-flasks, i.e. 100 g glucose l^–1; 1 g NH₄Cl l^–1; 2 g rice flour l^–1; 5 g KH₂PO₄ l^–1 and 2.5 g MgSO₄ l^–1, was obtained through a fractional factorial design and then scaled-up to a fluidized bioreactor. The effects of carbon and nitrogen concentrations, inoculum size, aeration, flow rate and bead sizes on batch bikaverin production using immobilized G. fujikuroi in a fluidized bioreactor were determined by an orthogonal experimental design. Concentrations of up to 6.83 g bikaverin l^–1 were obtained when the medium contained 100 g glucose l^–1 and 1 g NH₄Cl l^–1 with an inoculum ratio of 10% v/v, an aeration rate of 3 volumes of air per volume of medium min^–1, and a bead size of 3 mm. Based on dry weight, the bikaverin production was 30–100 times larger than found in submerged culture and approximately three times larger than reported for solid substrate fermentation.     

Accumulation,regulation and toxicity of copper,zinc, lead and mercury in Hyalella azteca

Zinc, lead and mercury accumulation in the amphipod Hyalella azteca increases with increasing exposure to metals. During 10 week chronic toxicity tests, metal accumulated at the highest non-toxic/lowest toxic concentration was 126/136 µg Zn g^–1, 7.1/16 µg Pb g^–1 and 56/90 µg Hg g^–1 dry weight. Concentrations of lead and mercyry in control animals were substantially lower (1.3 µg Pb g^–1 and 0.4 µg Hg g^–1), but concentrations of zinc in controls (74 µg g^–1) were about one half those of the lowest toxic concentration. Copper was completely regulated. Accumulated copper concentrations after 10 weeks exposure to all waterborne copper concentrations resulting in less than 100% mortality were not significantly different from controls (79 µg g^–1). Lead and mercury concentrations in wild H. azteca should be useful indicators of potential toxicity. Zinc accumulation may also be a useful indicator of zinc toxicity, but careful comparison with control or reference animals is necessary because of the small differences between toxic and control concentrations. Copper is not accumulated by H. azteca under chronic exposure conditions and body burdens of field animals cannot be used as an indicator of exposure or potential toxic effects. Short term exposures to copper, however, result in elevated copper concentrations in H. azteca, even at concentrations below those causing chronic toxicity. Short term bioaccumulation studies might, therefore, provide a useful indication of potential chronic copper toxicity.     

Optimisation of the cell release from immobilised cells of Bacillus simplex cultivated in culture media enriched with Cd2+: influence of Cd2+, inoculum size, culture medium and alginate beads characteristics

Calcium alginate beads were used to entrap a Bacillus sp. that has the ability to biosorb cadmium. During the batch incubation of alginate beads in a `rich' or a `poor' liquid medium, cell release out of the beads was noticed with a lag phase which was inversely proportional to the inoculum size (2×10⁷ or 2×10⁸ cells ml^–1 alginate), to the medium content, and proportional to the alginate concentration (10 or 15 g l^–1) and to the cadmium concentration (1, 5 or 10 mg l^–1). In addition, the cell release occurred more quickly when the medium was renewed. When the concentration was below 5 mg l^–1, the alginate matrix seemed to protect the bacteria against Cd²⁺ toxicity.     

Dynamics of bacterioplankton in a mesotrophic French reservoir (Pareloup)

Bacterioplankton abundance, biomass and production were studied at a central station (35 m depth) from April 1987 to September 1988 in a mesotrophic reservoir. Bacterial production was calculated by the (³H) thymidine method.For the water column, integrated estimates of bacterioplankton abundance ranged from 2.3 10⁹ to 4.6 10⁹ cells l^–1, and carbon biomass from 0.037 to 0.068 mg C l^–1; the thymidine incorporation rates ranged from 0.8 to 17.2 picomoles l^–1 h^–1, leading to net bacterial production estimates of less than 0.7 µg C l^–1 d^–1 in winter to 18 µg C l^–1 d^–1 in summer. About 55% of the production occurred in the euphotic layers.Over the year, the bacterial carbon requirement represented 90% of the autotrophic production for the whole lake. It was five times lower than autotrophic production in spring, but twice as high in summer. This important temporal lack of balance suggests that not all the spring primary production products are consumed immediately and/or that other carbon sources probably support bacterial growth in summer.     

Improvement of cephalomanine production in Taxus chinensis cells by a combination of sucrose and methyl jasmonate

Cell suspension cultures of Taxus chinensis, supplemented with 25 g sucrose l^–1, produced 11 mg cephalomanine l^–1, 21 g biomass l^–1 and 19 nkat geranylgeranyl diphosphate (GGPP) synthase activity g protein^–1. Supplementation of the cultures with 100 M methyl jasmonate (MJA) produced 17 mg cephalomanine l^–1, 6 g biomass l^–1 and 78 nkat GGPP synthase activity g protein^–1. Addition of sucrose and MJA together produced 24 mg cephalomanine l^–1, 18 g biomass l^–1 and 55 nkat GGPP synthase activity g protein^–1.     

Effects of sublethal exposure to copper on diel activity of sea catfish,Arius felis

The effect of sublethal exposure to copper on the diel activity of sea catfish, Arius felis, was examined after 72 h static exposure to copper (0.0, 0.05, 0.1, or 0.2 mg l^–1 ). The locomotor behavior of 28 individual fish was electronically monitored in a 16-compartment rosette tank. Activity was quantified by the number of entries into compartments per hour. Controls and copper-exposed fish (0.05 mg l^–1) behaved similarly after treatment and displayed no significant effects of handling or copper exposure on diel activity. However, fish exposed to 0.1 and 0.2 mg l^–1 copper were hyperactive immediately following exposure; these fish had significantly greater numbers of entries into compartments per hour for entire 24-h monitoring periods (P < 0.01) and for the 12 h of photophase (P < 0.001) and of scotophase (P < 0.001) than fish in the other two experimental groups. The normal diel activity cycle was abolished. Additionally, exposure of fish to copper at 0.1 and 0.2 mg l^–1 resulted in significantly less variability in activity (P < 0.05) compared to control fish and those exposed to 0.05 mg Cu l^–1.     

Plant Regeneration from Mesophyll Protoplasts of Echinacea Purpurea

An efficient plant regeneration system was developed from isolated protoplasts of Echinacea purpurea L. using an alginate block/liquid culture system. Viable protoplasts could be routinely isolated from young leaves of Echinacea seedlings in an isolation mixture containing 1.0% cellulase Onozuka R-10, 0.5% pectinase and 0.3 mol l^–1 mannitol. Purified protoplasts were embedded in 0.6% Na-alginate block at a density of 1 × 10⁵/ml and cultured in a modified MS medium containing 0.3 mol l^–1 sucrose, 2.5 µmol l^–1 BA and 5.0 µmol l^–1 2,4-D. Cell colonies were observed after 4 weeks of culture, and the protoplast-derived colonies formed calluses when transferred onto 0.25% gellan gum-solidified MS medium supplemented with 1.0 µmol l^–1 BA and 2.0 µmol l^–1 IBA. Shoot organogenesis from protoplast-derived callus was induced on MS medium supplemented with 5.0 µmol l^–1 BA and 2.0 µmol l^–1 IBA. Complete plantlets were obtained from the regenerated shoots on MS basal medium. The protoplast to plant regeneration protocol developed in this study provides the prerequisite for creating novel genotypes of this valuable medicinal species through genetic manipulation.     

2-Propanol degradation by Sulfolobus solfataricus

Sulfolobus solfataricus used 2-propanol and 2-propanone (acetone) when grown in static cultures at 78 °C with or without glucose at 10 g l^–1. The presence of 3.92 g 2-propanol l^–1 in both cases inhibited growth. However, acetone accumulation following 2-propanol depletion suggested that 2-propanol was co-metabolized via the acetone metabolic pathway. Glucose at 10 g l^–1 increased 2-propanol and acetone utilization from 0.93 g l^–1 to 1.77 g l^–1 and from 0.11 g l^–1 to 1.62 g l^–1, respectively. Without glucose, immobilized S. solfataricus cells increased the 2-propanol removal rate to 0.035 g l^–1 h^–1, compared to 0.0012 g l^–1 h^–1 by its suspended counterpart. The results suggest the establishment of an immobilized reactor configuration is preferential for the treatment of high temperature solvent waste streams by this acidothermophilic Crenarchaeon.