Effects of copper enrichment on survival,growth and photosynthetic pigment of seedlings and young plants of the eelgrass Zostera marina

Copper (Cu²⁺) is an essential nutrient for plants but toxic at high concentrations. We subjected seedlings and young plants of eelgrass Zostera marina to different seawater Cu concentrations (3, 4, 5, 10, 30 and 50?µg?l^?1) for over 30 days under controlled laboratory conditions. Natural seawater without added Cu (3?µg?l^?1) was used as reference seawater. We measured plant response in terms of survivorship, morphology, growth, productivity and leaf pigment concentration. Survival analysis combined with morphological, dynamic and productive assessment suggested that the optimum seawater Cu concentration for the establishment of Z. marina seedlings and young plants is 4?μg?l^?1. The photosynthetic response of young plants to copper enrichment, including an increase in chlorophyll content under low Cu concentration treatment but significant decrease when treated with high concentrations of Cu, is similar to those reported for other seagrass species. NOEC (no observed effect concentration), LOEC (lowest observed effect concentration) and LC₅₀ (lethal concentration that caused an increase in mortality to 50% of that of the control) values of seedlings were significantly lower than those of young plants, implying a reduced Cu tolerance to high concentrations (>10?μg?l^?1). This study provides data that could prove helpful in the development of successful eelgrass restoration and conservation.     

The plasma zinc/serum copper ratio as a biomarker in children with autism spectrum disorders

The frequency of zinc deficiency, copper toxicity and low zinc/copper in children with autism spectrum disorders (ASDs) may indicate decrement in metallothionein system functioning. A retrospective review of plasma zinc, serum copper and zinc/copper was performed on data from 230 children with autistic disorder, pervasive developmental disorder-NOS and Asperger’s syndrome. The entire cohort’s mean zinc level was 77.2 μg dl^?1, mean copper level was 131.5 μg dl^?1, and mean Zn/Cu was 0.608, which was below the 0.7 cut-off of the lowest 2.5% of healthy children. The plasma zinc/serum copper ratio may be a biomarker of heavy metal, particularly mercury, toxicity in children with ASDs.     

Life-history traits of a tube-dwelling corophioid amphipod, Paracorophium excavatum, exposed to sediment copper

This is the first demonstration that sediment contaminants can influence the reproduction of amphipods. Groups of Paracorophium excavatum from a slightly contaminated estuarine site were held within laboratory mesocosms containing four copper-spiked estuarine sediments (Cu 14-46 μg g⁻¹ dry weight) and a control sediment (Cu 5 μg g⁻¹ dry weight) at 15 °C for 28 days. Copper sediment concentration did not affect the amphipod sex ratio. Female maturation was inhibited within copper-spiked sediments but female length was similar. Juvenile recruitment occurred only in sediments containing less than 20 μg g⁻¹. Males were significantly larger than females in the control sediment (Cu 5 μg g⁻¹ dry weight) and male length decreased linearly with increasing copper concentration. The copper concentration within whole body tissues increased with dry body dry weight in all sediments except the highest copper concentration. Following 28 days of exposure, none of the female amphipods from the copper-dosed sediments was brooding embryos. In contrast, brood size of females in the control sediment (Cu 5 μg g⁻¹) was similar to field samples. Because low concentrations of sediment copper affect the maturation and growth rates of male and female amphipods differently, these life-history traits could affect the population structure of amphipods exposed to copper contaminated sediments.     

Production of juvenile Atlantic salmon, Salmo salar L., and brown trout, Salmo trutta L., within different sections of a small enriched Norwegian river

Growth, density and production of juvenile Atlantic salmon and brown trout were studied in three different sections of the Kvassheimsåna River in south-western Norway from 1979 to 1983. Section 1. in the upper part of the river, is located above a waterfall impassable for migratory salmonids and is surrounded by grazing land. Sections 2 and 3, in the middle and lower parts of the river, are influenced by agricultural activity. Total nitrogen concentration varied between 250 and 1000 μg l ^?1 in section 1 and 1500 and 2500 μg l^?1 in sections 2 and 3. Total phosphorus (Tot-P) concentrations also increased with decreasing altitude: 19–46 μg l^?1 in section I and 31–101 μg l ^?1 in sections 2 and 3. The number of 0 + salmon in sections 2 and 3 varied between 30.1 and 167.8 specimens 100 m ^?2, with means 90.2 and 95.2 specimens 100 m ^?2:, respectively; the density of 1 + salmon, with mean values of 16.3 and 51.0 specimens 100m^?2 was significantly correlated with the original fry density. The growth rate of 0+ salmon was not inversely related to cohort density, but was significantly so for 1 + salmon. Mean annual salmon production in section 2 was 1595 g 100 m^?2 year ¹, and in section 3 was 841 g 100m^?2 year ¹. A logarithmic function gave the best curve fit between salmon production and mean annual biomass. Thus, production levelled off for the highest values recorded in section 2, and perhaps approached the carrying capacity of the stream. A multiple regression analysis showed that yearly variation in 1 + salmon density was the single factor accounting for most of the total variability in production (60%). Variation in water temperature and nutrient content were not significantly related to variation in fish production. Densities of brown trout were low in all sections (<20 specimens 100m ^?2). Fry density was highest in section 3 and parr density in section 1. All age groups of sympatric brown trout grew significantly faster in sections 2 and 3 compared with allopatric brown trout in section 1.     

Effects of stocking density on growth and skin color of juvenile darkbarbel catfish Pelteobagrus vachelli (Richardson)

A 3‐month experiment was conducted to investigate the effects of stocking density on growth performance and skin color of juvenile darkbarbel catfish. Experimental fish (5.0 ± 0.6 g; n = 160) were stocked in triplicate in 2 m² concrete tanks in a greenhouse at the initial density of 50, 150, 400 and 650 fish m^?2, respectively (0.42, 1.25, 3.33 and 5.42 kg m^?3). At the end of the experiment, 60 fish from each tank were sampled to record body weight and total length, and six fish from each tank were sampled to measure skin color by instrumental color analysis. Results showed that stocking density affected growth performance significantly (P < 0.05). Final mean weight (W), final mean total length (L) and specific growth rate (SGR) were significantly higher at 150 fish m^?2 treatment and lower in both the highest and lowest (650 and 50 fish m^?2) density treatments (P < 0.05). The condition factor (CF) was higher in 150 and 50 fish m^?2 and lowest at 650 fish m^?2 treatment; the coefficient of variation of weight (CV) tended to increase with the increase in stocking density, which was lowest in 50 fish m^?2 and highest in 650 fish m^?2. Stocking density also affected the skin color parameters, L* (lightness), a* (redness), b* (yellowness) and h* (hue), significantly (P < 0.05). The values of these parameters tended to decline with the increase in stocking density. Results of the present study suggest that growth performance and skin color responses to stock density exhibit different patterns for darkbarbel catfish juveniles: highest and lowest densities impaired growth rate, and higher density had a darkening effect on skin color.     

Influence of available dietary carbohydrate content on tolerance of waterborne copper by rainbow trout,Salmo gairdneri Richardson*

The effect of increased available dietary carbohydrate on the tolerance of waterborne copper by rainbow trout was investigated. Trout were reared for six to eight weeks on one of four isocaloric and isonitrogenous diets which varied in available carbohydrate content due to either processing (extrusion v. steam-pelleting) or supplementation with cerelose (0 or 21%). The incipient lethal level (ILL) of copper for trout reared on the low-available-carbohydrate steam-pelleted diet was 350 μg I^?1, significantly higher than the 276 μg I^?1 shown for trout reared on the high-available-carbohydrate extruded diet. The ILL of copper for the low-carbohydrate group (0% cerelose) was 408 μg I^?1, significantly higher than the ILL of 246 μg I^?1 of trout reared on the high-carbohydrate diet (21% cerelose). Increased available dietary carbohydrate resulted in increased liver glycogen content and liver-body weight ratios and decreased liver protein content. Significant correlations were evident between waterborne copper tolerance and liver glycogen, liver-body weight ratios and liver protein. Relative to those on high carbohydrate diets, livers of fish on low-carbohydrate diets contained higher concentrations of copper after seven days of exposure to 0 or 196 μg I^?1 of copper. The elevated liver glycogen levels, resulting from increased dietary carbohydrate, are thought to have impaired liver function, thus reducing copper tolerance.     

ADSORPTION AND UPTAKE OF COPPER BY THE GREEN ALGA SCENEDESMUS SUBSPICATUS (CHLOROPHYTA)1

Copper (II) accumulation has been investigated in the green alga Scenedesmus subspicatus G. Brinkmann considering both adsorption and uptake kinetics. Experiments were conducted in a Cu- and PH-buffered medium at different free Cu²⁺ concentrations that were neither growth limiting nor toxic. We distinguished between adsorption on the cell surface and intracellular uptake by extracting copper from the cells with EDTA. Data from short-term experiments were compared with data obtained from experiments under steady state conditions. The accumulation of Cu can be described by two processes, an initial fast adsorption occurring within a minute followed by a slower intracellular uptake. Metal uptake followed Michaelis-Menten kinetics and is mediated by two systems, one with low and the other with high affinity. The maximum uptake rates (1.30 × 10^?-¹⁰ mol·[g dry wt algae]^?1· min^?1, 3.67 × 10^?-⁹ mol·[g dry wt algae]^?1·min^?1), and the half-saturation constants (6.84 × 10^?-¹⁴ M, 2.82 × 10^?-¹² M) for the two uptake systems were determined using the Lineweaver-Burk plot. The calculated maximum concentration of binding sites on the surface of the algae is initially higher (9.0 × 10^?-⁶ mol Cu.[g dry wt algae]^?1) than under steady state conditions (2.9 × 10^?-⁶ mol Cu·[g dry wt algae]^?1). This suggests that the initial binding to the algal surface comprises the binding to specific transport ligands as well as to inert adsorption sites. The conditional stability constant of the Cu binding to surface ligands was calculated as log K_Cu= 11.0 at pH 7.9. This freshwater alga has a high ability to accumulate Cu, reflecting its adaptation to the bioavailable concentration of copper.     

Assessment of heavy metal accumulation in Anzali wetland,Iran, using a submerged aquatic plant,Ceratophyllum demersum

The objective of this study in 2009 was to examine whether levels of cadmium (Cd), copper (Cu), zinc (Zn), lead (Pb) and chromium (Cr) were higher in the leaves than in the stems of a submerged aquatic plant Ceratophyllum demersum in Anzali wetland. Cadmium, Pb and Cr concentrations were highest in the leaves. The mean concentrations of Cd and Cr in the leaves at all the sampling sites ranged between 0.94–1.26 μg g^?1 and 1.03–2.71 μg g^?1, respectively. Lead also had its highest concentrations in the leaves. The mean concentration of Pb in the leaves at all sampling sites ranged between 7.49–11.88 μg g^?1. Copper and Zn concentrations were highest in the stems. The mean concentrations of Cu and Zn in the stems at all sampling sites ranged between 10.79–17.91 μg g^?1 and 19.89–40.01 μg g^?1, respectively. Cadmium and Pb concentrations were higher in the leaves than in the stems, while Zn concentration was higher in the stems than in the leaves. Accumulation of Cu and Cr in the organs of C. demersum was in descending order of leaf ～ stem, since there was no significant difference between their mean concentrations in the leaves and stems.     

Effects of copper and other metals on fertilization,embryo development,larval survival and settlement of the polychaete Nereis (Neanthes) virens

Summary

Nectochaete larvae of the ecologically and economically important ragworm, Nereis virens, were exposed to cadmium, chromium, copper, lead and zinc dissolved in seawater to nominal concentrations ranging from 0 to 5000 μg l^?1. Copper was the most toxic (mean LC50 of 76.5 μg l^?1 ± 95% CI 73.8–79.2 after 96 h exposure) and so was used for subsequent experiments. Exposure of gametes to greater than 500 μg l^?1 copper for 2 or 4 h at 10°C prior to fertilization, or a 10 min exposure during fertilization, significantly reduced embryo developmental success. The effect of copper on larval settlement was also assessed using sediment spiked to a range of concentrations (0, 50, 250, 500, 1000 mg kg!1 dry weight). Significantly fewer larvae were found in sediment of $250 mg kg!1 in comparison to the control or the 50 mg kg!1 treatment. Assessment of living larvae also confirmed a significant reduction in settlement, but in all treatments compared to the control, although the number of dead larvae also increased as the concentrations increased. These effects may have important implications for reproductive success and recruitment of N. virens to polluted sediments.     

Inactivation of Cronobacter spp. (Enterobacter sakazakii) in infant formula using lactic acid,copper sulfate and monolaurin

Aims: To investigate the effect of lactic acid (LA), copper (II), and monolaurin as natural antimicrobials against Cronobacter in infant formula. Methods and Results: The effect of LA (0·1, 0·2 and 0·3% v/v), copper (II) (10, 50 and 100 μg ml^?1) and monolaurin (1000, 2000, and 3000 μg ml^?1) suspended into tween‐80? or dissolved in ethanol against Cronobacter in infant formula was investigated. Reconstituted infant formula and powdered infant formula were inoculated with five strains of Cronobacter spp. at the levels of c. 1 × 10⁶ CFU ml^?1 and 1 × 10³ CFU g^?1, respectively. LA at 0·2% v/v had a bacteriostatic effect on Cronobacter growth, whereas 0·3% v/v LA resulted in c. 3 log₁₀ reduction. Copper (II) at the levels of 50 μg ml^?1 and 100 μg ml^?1 elicited c. 1 and 2 log₁₀ reductions, respectively. The combination of 0·2% LA and 50 μg ml^?1 copper (II) resulted in a complete elimination of the organism. Monolaurin exhibited a slight inhibitory activity against Cronobacter (c. 1·5 log₁₀ difference) compared to the control when ethanol was used to deliver monolaurin. Conclusions: A complete elimination of Cronobacter was obtained when a combination of sublethal concentrations of LA (0·2%) and copper (II) (50 μg ml^?1) was used. Significance and Impact of the Study: The use of the synergistic interactive combination of LA and copper (II) could be beneficial to control Cronobacter in the infant formula industry.     

Changes in vulnerability of Daphnia to an insecticide application depending on the population phase

1. Laboratory populations of cloned Daphnia pulex at three population phases (growing phase, density peak, declining phase) were exposed to the insecticide carbaryl at 15 μg L^?1, which killed juveniles but not adults. 2. The population decline was largest and recovery was slowest when the chemical was applied at the density peak, indicating that the population was most sensitive to the chemical at that stage. Populations in the declining phase were least affected by the treatment. 3. We propose that food condition, which varies with population density, was a major factor affecting vulnerability to carbaryl. In that case, Daphnia populations in lakes would be most vulnerable to toxic chemicals during food‐limited condition such as the spring clear‐water phase.     

EFFECT OF CYSTEINE AND METHIONINE ON SULFATE UPTAKE AND PRIMARY PRODUCTIVITY BY AXENIC ALGAL CULTURES AND LAKE MICROPLANKTON COMMUNITIES1

The presence of up to 500 μg sulfur·l^?1 of an equimolar mixture of cysteine and methionine had virtually no effect on the SO₄^2- uptake rate of Navicula pelliculosa, (Bréb.) Hilse whereas the rate of Ankistrodesmus falcatus (Corda) Ralfs was decreased by the presence of 500 μg S· l^?1 and Anabaena flos-aquae (Lyngbye) Bréb. by 50 μg S·l^?1. Primary productivity in these axenic cultures was affected (decreased) only in A. falcatus. The C:S uptake ratio was lowest in N. pelliculosa and highest in A. falcatus. Considering these species as representative of groups of naturally occurring algae, patterns of SO₄^2- uptake and primary productivity in a eutrophic and a moderately oligotrophic lake reflected the results of the algal culturing experiments: SO₄^2- uptake rates, relative to primary productivity, were higher in the presence of diatoms and bluegreen algae and lower when green algae were present; the addition of the cysteine I methionine mixture to the lake waters decreased the rate of microplankton SO₄^2- uptake in correlation with the makeup of the algal community; primary productivity decreased upon the addition of cysteine I methionine when green algae were relatively abundant. It is concluded that, in most fresh water systems, the effects of organic sulfur pollution on algal SO₄^2- uptake and primary productivity are insignificant as compared to other ecological changes that occur due to that pollution.     

The geographic variation and potential risk of DDT in the blood of Pied Kingfishers from northern KwaZulu-Natal,South Africa

Evans, S.W. & Bouwman, H. 2000. The geographic variation and potential risk of DDT in the blood of Pied Kingfishers from northern KwaZulu-Natal, South Africa. Ostrich 71 (1 & 2): 351–354.

DDT has, since 1946 been used in the intradomicilliary control of malaria in northern KwaZulu-Natal. The Pied Kingfisher was selected as representative for organisms in relatively high trophic levels. Blood was obtained from Pied Kingfishers at Kosi Bay (n = 5), Pongolo Floodplain (n = 13), Mkuzi Nature Reserve (n = 4), Ndumu Nature Reserve (n = 4) and St Lucia (n = 3), extracted and analysed [SWEl] via gas chromatography. The highest blood DDE and σDDT concentrations were obtained for the birds from the Pongolo Floodplain (means of 95.92 μg 1^?1/107.01 μg μg 1^?1) and Kosi Bay Nature Reserve (means of 189.09 μg 1^?l/241.8 μg 1^?1). DDT was detected in the blood of Pied Kingfishers from Kosi Bay (mean 47.14 μg 1^?1) and Pongolo Floodplain (mean 44.34 pg 1^?1) only. This indicated their proximity to DDT application and the greater influx of DDT and its metabolites into the water component of these systems. The EDDT plasma concentrations in the Pied Kingfisher blood were calculated by multiplying the blood values of σDDT by 1.8. Using the regression, log₁₀Y = 0.7785 + 0.8593 (log₁₀X), relating the σDDT in eggs to σDDT in plasma of American Kestrel Falco sparverius it was possible to calculate the mean Pied Kingfisher egg σDDT concentration. The approximate mean Pied Kingfisher egg concentration of σDDT was calculated at 2.26 mg kg^?1 for Kosi Bay and 1.24 mg kg^?1 for the Pongolo Floodplain. Using the highest calculated plasma value of σDDT for Kosi Bay and the Pongolo Floodplain indicated that egg σDDT concentrations could be as high as 4.01 mg kg^?1 and 4.17 mg kg^?1 respectively. These calculated levels may be significant when compared to levels of DDE, known to have a detrimental effect, in the eggs of the Brown Pelican Pelecanus occidentalis, where a concentration of 2.5 to 3 mg kg^?1 was associated with substantially impaired reproductive success. The highest calculated egg concentration was nearing this level and it is therefore possible that the Pied Kingfisher population may be at risk.     

NaCS‐PDMDAAC immobilized cultivation of recombinant Dictyostelium discoideum for soluble human Fas ligand production

Dictyostelium discoideum is a promising eukaryotic host for the expression of heterologous proteins requiring post‐translational modifications. However, the dilute nature of D. discoideum cell culture limits applications for high value proteins production. D. discoideum cells, entrapped in sodium cellulose sulfate/poly‐dimethyl‐diallyl‐ammonium chloride (NaCS‐PDMDAAC) capsules were used for biosynthesis of the heterologous protein, soluble human Fas ligand (hFasL). Semi‐continuous cultivations with capsules recycling were carried out in shake flasks. Also, a scaled‐up cultivation of immobilized D. discoideum for hFasL production in a customized vitreous airlift bioreactor was conducted. The results show that NaCS‐PDMDAAC capsules have desirable biophysical properties including biocompatibility with the D. discoideum cells and good mechanical stability throughout the duration of cultivation. A maximum cell density of 2.02 × 10⁷ cells mL^?1 (equivalent to a maximum cell density of 2.22 × 10⁸ cells mL^?1 in capsules) and a hFasL concentration of 130.40 μg L^?1 (equivalent to a hFasL concentration of 1434.40 μg L^?1 in capsules) were obtained in shake flask cultivation with capsules recycling. Also, a maximum cell density of 1.72 × 10⁷cells mL^?1 (equivalent to a maximum cell density of 1.89 × 10⁸ cells mL^?1 in capsules) and a hFasL concentration of 106.10 μg L^?1 (equivalent to a hFasL concentration of 1167.10 μg L^?1 in capsules) were obtained after ～170 h cultivation in the airlift bioreactor (with a working volume of 200 mL in a 315 mL bioreactor). As the article presents a premier work in the application of NaCS‐PDMDAAC immobilized D. discoideum cells for the production of hFasL, more work is required to further optimize the system to generate higher cell densities and hFasL titers for large‐scale applications. © 2014 American Institute of Chemical Engineers Biotechnol. Prog., 31:424–430, 2015     

GROWTH AND MORPHOLOGY OF YOUNG GAMETOPHYTES OF PORPHYRA ABBOTTAE (RHODOPHYTA): EFFECTS OF ENVIRONMENTAL FACTORS IN CULTURE1

Growth, blade shape and blade thickness of young gametophytes of Porphyra abbottae Krishnamurthy cultured from conchospores were determined at various combinations of temperature (8, 10, 12° C), photon flux density (17.5, 70, 140 μmol·m-^?2·S^?1), nutrient concentration (5, 25, 50, 100% f medium) and water motion (0, 50, 100, 150 rpm). Growth (as surface area) was light-saturated at 70 μmol· m^?2· S^?1, light-inhabited at 140 μmol·m^?2· S^?1, and nutrient-saturated an 25% f medium. Temperature had no significant effect on growth. Water motion and nutrients had an interactive effect on growth, with water motion having the greatest effect at the lowest nutrient concentrations. Water motion enhanced growth even at saturating nutrient concentrations. Blade length / width ratio was greater in low light (2.5) than in saturating light (1.9); with increasing water motion the ratio increased from 1.2 to 2.4. Blade thickness (53-88 μm) was greatest at the highest nutrient concentrations and at the lowest water motion levels. Temperature and light did not have a consistent effect on blade thickness.     

Response of juvenile tench Tinca tinca (L.) to the anaesthetic 2-phenoxyethanol

The response of tench Tinca tinca aged 40–171 days post‐hatch (22–49 mm TL) to the anaesthetic 2‐phenoxyethanol was studied at 25°C. The lowest effective concentration of 2‐phenoxyethanol increased with age, while the highest safe concentration decreased. The fish aged 40 days post‐hatch required a significantly (P ≤ 0.05) shorter time to become anaesthetized than did older fish. The recovery time after 15 min of exposure to 2‐phenoxyethanol at 0.45 g dm^?3 was significantly shorter in the 40‐day‐old fish than in older fish. In juveniles of the same age, induction time or recovery time did not depend on their size or condition (Fulton's coefficient). At 25°C, 2‐phenoxyethanol at 0.5 g dm^?3 may be used to efficiently and safely anaesthetize T. tinca juveniles.     

Lead Crystal: An Important Potential Source of Lead Exposure

Abstract

We have examined the potential of alcoholic beverages to elute Pb from crystal wine decanters and glasses. Port wine containing 89 Pb L^?1 was aliquoted into three lead crystal wine decanters. The wine Pb content rose steadily to a mean of 3,518 μg L^?1 after 4 months. We then analyzed wines and liquors which had been stored in crystal decanters in the homes of our colleagues for periods of six months or longer. In the first four homes, eight decanters contained beverages with a mean Pb concentration of 7,781μg L^?1 In a short-term experiment, white wine was found to elute small quantities of Pb from lead crystal glasses within minutes; the mean wine Pb concentration rose from 33 μg L^?1 at time zero to a mean of 99 μg L^?1 after four hours. We conclude that, in particular, lead crystal wine decanters are potentially an important source of Pb exposure for a segment of the population. In addition, lead crystal baby bottles, although not as widely available, were found to elute Pb into apple juice and infant formula; their sale should be banned.     

Comparative effects of adenine analogs upon metabolic cooperation between Chinese hamster cells with different levels of adenine phosphoribosyltransferase activity

Colony formation by variant Chinese hamster cells highly resistant to adenine analogs and deficient in adenine phosphoribosyltransferase (APRT) activity was measured after co-cultivation with APRT⁺, CHO-K1 cells in medium containing one of three different adenine analogs. Depending upon the density of APRT⁺ cells and the specific adenine analog, large differences in the recovery of APRT^? colonies were observed. The particular adenine analog and APRT⁺ cell density were more significant factors in the recovery of APRT^? colonies than the concentration of the analog or the level of APRT activity. The number of wild-type cells (CHO-K1) required to inhibit formation of APRT^? colonies by 50% (mean lethal density; MLD₅₀) with 65 μg/ml 8-aza-adenine (AzA) as the selective drug was 8.0 × 10⁵ cells/100 mm dish (1.5 × 10⁴/cm²). With 100 μg/ml 2,6-diaminopurine (DAP) the MLD₅₀ for CHO-K1 was 4.0 × 10⁵ cells/100 mm dish (7.3 × 10³/cm²). The MLD₅₀ for CHO-K1 when the DAP concentration was decreased to 50 μg/ml was only slightly higher, 5 × 10⁵ cells/100 mm dish (9.1 × 10³/cm²). The most toxic effect was observed with 2-fluoroadenine (FA). The MLD₅₀ for CHO-K1 in 2 μg/ml FA was 4.5 × 10⁴ cells/100 mm dish (8.2 × 10²/cm²), a cell density which permits minimal direct contact between APRT⁺ and APRT^? cells. The toxic effects of FA on individually resistant, APRT^? cells were found to be mediated by metabolites released into the medium by dying APRT⁺ cells. This metabolite toxicity to APRT^? cells was also demonstrated in mixtures with cells having only 8% of wild-type APRT activity. The MLD₅₀ for these APRT⁺ (8%) cells in 2 μg/ml FA was 7.5 × 10⁴ cells/100 dish (1.4 × 10³/cm²), a small difference from the MLD₅₀ for cells with wild-type levels of APRT activity. The differences in the recovery of APRT^? colonies from mixtures with APRT⁺ cells in these three adenine analogs are critical to the design of procedures for the selection of APRT^? cells from populations of APRT⁺ cells and emphasize the importance of establishing the parameters of metabolic cooperation, not only in terms of cell density but also with regard to the particular selective agent, in any experiment designed to determine precise mutation rates or to test putative mutagens upon mammalian cells in culture.     

The effects of copper on photosynthesis and biomolecules yield in Chlorolobion braunii

Our knowledge of the effects of copper on microalgal physiology is largely based on studies conducted with high copper concentrations; much less is known when environmentally relevant copper levels come into question. Here, we evaluated the physiology of Chlorolobion braunii exposed to free copper ion concentrations between 5.7 × 10^?9 and 5.0 × 10^?6 mol · L^?1, thus including environmentally relevant values. Population growth and maximum photosynthetic quantum yield of PSII were determined daily during the 96 h laboratory controlled experiment. Exponentially‐growing cells (48 h) were analyzed for effective quantum yield and rapid light curves (RLC), and total lipids, proteins, carbohydrates, chlorophyll a and carotenoids were determined. The results showed that growth rates and population density decreased gradually as copper increased in experiment, but the photosynthetic parameters (maximum and effective quantum yields) and photochemical quenching (qP) decreased only at the highest free copper concentration tested (5.0 × 10^?6 mol · L^?1); nonphotochemical quenching (NPQ) increased gradually with copper increase. The RLC parameters Ek and rETRmax were inversely proportional to copper concentration, while α and Im decreased only at 5.0 × 10^?6 mol · L^?1. The effects of copper in biomolecules yield (mg · L^?1) varied depending on the biomolecule. Lipid yield increased at free copper concentration as low as 2.5 × 10^?8 mol · L^?1, but proteins and carbohydrates were constant throughout.     

Extracts and phenolic compounds from Zuccagnia punctata as fungicide seed protectants for corn

The ethanolic leaf extract (TZP) of Zuccagnia punctata, its ethereal fraction (Eet), 2′,4′‐dihydroxychalcone (DC), 2′,4′‐dihydroxy‐3′‐methoxychalcone (DMC) and 7‐hydroxy‐3′,4′‐dimethoxyflavone (HF) were evaluated as fungicide seed protectants on corn. Microdilution assays on a set of Fusarium strains showed minimum inhibitory concentrations (MICs) of 400–800 μg mL^?1 (TZP), 50–100 μg mL^?1 (Eet), 25–50 μg mL^?1 (DC), 50–100 μg mL^?1 (DMC) and 200–400 μg mL^?1 (HF), with minimum fungicidal concentration (MFC)/MIC = 1. Suspensions of TZP, Eet, DC and DMC at MIC × 20 incorporated to the grains at rates of 1920, 240, 120 and 240 mg dry matter kg^?1 of grain, respectively, increased the elongation of the primary roots (24–44%) and the number of seminal roots (44–50%). TZP also increased the number of secondary roots. HF was phytotoxic. Suspensions of TZP, Eet, DC and DMC suppressed the endogenous grain mycoflora at levels similar to those recorded for a thiram + carbendazim‐based fungicide. Grains treated with TZP (1920 mg kg^?1), Eet (240 mg kg^?1), DC (120 mg kg^?1) and DMC (240 mg kg^?1) stimulated the growth of the seedling root system both because of fungal suppression and hormetic effects in greenhouse curative and preventive assays against Fusarium verticillioides on a sand/soil substrate. Eet and its chalcones also reduced the severity of seedling blight more than the thiram + carbendazim‐based fungicide in preventive assays and led to the same disease severity observed for the fungicide treatment in the curative assays. Our results show that Eet and its chalcones not only were effective seed protectants against F. verticillioides and other seedborne fungi, but also improved the early performance of maize seedlings.