Influence de divers facteurs écologiques sur la bioaccumulation d'éléments métalliques (Cd, Cu, Pb, Zn) chez de jeunes palourdes (Ruditapes philippinarum) au cours du prégrossissement en nourricerie

La bioaccumulation des métaux chez les mollusques peut être une conséquence de la désorption des éléments métalliques fixés sur les particules inertes ou vivantes en suspension dans l'eau qui leur servent de nourriture. Les meilleures expériences réalisées jusqu'ici semblent indiquer que les matières en suspension ont un rôle mineur dans la contamination des mollusques. Mais quelles que soient les précautions prises, les expériences ne simulent jamais parfaitement les phénomènes naturels. Aussi, nous avons choisi de reprendre cette question à l'aide d'une expérience en grand volume où le nombre de paramètres contrôlés est restreint mais où les organismes sont dans des conditions aussi proches que possible de la réalité.Au cours d'un prégrossissement expérimental, les jeunes palourdes reçoivent des quantités variables contrôlées de nourriture phytoplanctonique. La fourniture d'une nourriture plus abondante a pour conséquence d'augmenter les quantités de cuivre et de zinc et au contraire de diminuer celles de plomb contenues dans chaque individu. Elle entraîne également une diminution des concentrations en cadmium et plomb traduisant une « dilution biologique » de ces métaux. L'augmentation de la densité de la population expérimentale a un effet négatif sur les quantités de cuivre et de zinc contenues dans les individus. A âge identique, les individus les plus grands présentent des concentrations plus faibles en cuivre et zinc et plus élevées en cadmium et plomb.L'utilisation des eaux marines souterraines présente deux avantages: une production algale intense à un coût économique négligeable et la possibilité par échange thermique de réchauffer en hiver les eaux marines naturelles et ainsi de maintenir une croissance des mollusques toute l'année. L'emploi des eaux de forage n'entraîne aucun effet néfaste dans les phénomènes de bioaccumulation des métaux, du moins pour ceux étudiés ici : cadmium, cuivre, plomb et zinc.Metal bioaccumulation in bivalves may occur as a consequence of the ingestion of inert or living particles with fixed trace elements. The best experiments like those carried out by BORCHARDT (1983, 1985) concerning Cd in mussels have shown that the role suspended matter plays in the contamination of molluscs is insignificant. But, however reliable the experimental methodologies, laboratory conditions never reproduce perfectly natural phenomena. We planned therefore to restudy the problem by using a largescale experiment where the procedure was characterized by a restricted number of controlled parameters and the organisms as close as possible to the real conditions.During an experimental nursing of young carpet shells, we assessed the influence of various controlled quantifies of phytoplankton and of experimental population density on the transfer of metal from their environment to molluscs.Carpet-shell brood (Ruditapes philippinarum) was distributed in several cylindrical containers the bottom of which consisted in a sieve. Food and seawater were renewed continuously by means of an ascending current (BAUD et BACHER, 1990). Nursing assays were carried out during summer over a period of 74 days. Eight groups of carpet shells were constituted according to food supplies (0, lx, 2x and 4x of Skeletonema costatum grown upon underground seawater plus natural phytoplankton) and population density (25 000 or 50 000 individuals per experimental container). Young molluscs were fed according to a cycle of 3 h-feeding periods and 2 h-periods with no food alternately. The average concentrations of algal cells in mollusc breeding seawater were 17.5, 35 and 70.103 cells/L. This seawater was renewed at a flow rate of 3 m3/h.At the end of the nursing period, molluscs exposed to different experimental conditions were separated by using sieves of different mesh-size (6, 8 and 10 mm). Young carpet shells were purged for 36 h in order to limit the overvaluation of bioaccumulated metal levels due to ingested matter (AMIARD-TRIQUET et al., 1984; KENNEDY, 1986). In each experimental and size-related categories, 90 individuals were sampled and divided into 3 groups of 30 specimens.In these groups, soft tissues were separated from the shells and oven-dried at 80 °C for 48 h. The dry samples were powdered and three aliquot parts of about 100 mg each were digested with 1 ml of concentrated nitric acid (HNO3, Suprapur) at 95 °C for 1 h. Then the trace element analyses were performed in this solution diluted with deionized water by dame (Zn) or by flameless (Cd, Cu, Pb) atomic absorption spectrophotometry using the Zeeman effect (AMIARD et al., 1987).The influence of both food supplies and experimental population density on the dry weight of soil tissues of young carpet shells, their metal concentration and body burdens were examined by means of multi-linear regression analysis.Increasing body burdens of Cu (4) and Zn (5) and decreasing body burden of Pb (3), corresponded to more abundant food supplies. The increase of phytoplankton supplies induced a decrease of Cd (6) and Pb (7) concentrations as a consequence of a « biological dilution » of these metals. Increasing density induced a depletion of Cu (4) and Zn (5) body burdens. Among individuals of the same age, the biggest ones exhibited the lowest concentrations of Cu and Zn (8 and 9) and the highest concentrations of Cd and Pb (6 and 7).Increased food supplies induce a biological dilution of Cd and Pb in young carpet-shells. These results are in agreement with previous data concerning Cd, Cu, Pb and Zn in different species (MACKAY et al., 1975; BOYDEN, 1971; PHELPS et al., 1985; BERTHET, 1986). Thus front a sanitary point of view, the use of ground seawater for algal culture is not a risk since metal concentrations in molluscs are not enhanced

Spatial variation of trace metals within intertidal beds of native mussels (Mytilus edulis) and non-native Pacific oysters (Crassostrea gigas): implications for the food web?

Abstract Pollution is of increasing concern within coastal regions and the prevalence of invasive species is also rising. Yet the impact of invasive species on the distribution and potential trophic transfer of metals has rarely been examined. Within European intertidal areas, the non-native Pacific oyster (Crassostrea gigas) is becoming established, forming reefs and displacing beds of the native blue mussel (Mytilus edulis). The main hypothesis tested is that the spatial pattern of metal accumulation within intertidal habitats will change should the abundance and distribution of C. gigas continue to increase. A comparative analysis of trace metal content (cadmium, lead, copper and zinc) in both species was carried out at four shores in south-east England. Metal concentrations in bivalve and sediment samples were determined after acid digestion by inductively coupled plasma-optical emission spectrometry. Although results showed variation in the quantities of zinc, copper and lead (mg m-2) in the two bivalve species, differences in shell thickness are also likely to influence the feeding behaviour of predators and intake of metals. The availability and potential for trophic transfer of metals within the coastal food web, should Pacific oysters transform intertidal habitats, is discussed

G-quadruplex formation at the 3′ end of telomere DNA inhibits its extension by telomerase, polymerase and unwinding by helicase

Telomere G-quadruplex is emerging as a promising anti-cancer target due to its inhibition to telomerase, an enzyme expressed in more than 85% tumors. Telomerase-mediated telomere extension and some other reactions require a free 3′ telomere end in single-stranded form. G-quadruplex formation near the 3′ end of telomere DNA can leave a 3′ single-stranded tail of various sizes. How these terminal structures affect reactions at telomere end is not clear. In this work, we studied the 3′ tail size-dependence of telomere extension by either telomerase or the alternative lengthening of telomere (ALT) mechanism as well as telomere G-quadruplex unwinding. We show that these reactions require a minimal tail of 8, 12 and 6 nt, respectively. Since we have shown that G-quadruplex tends to form at the farthest 3′ distal end of telomere DNA leaving a tail of no more than 5 nt, these results imply that G-quadruplex formation may play a role in regulating reactions at the telomere ends and, as a result, serve as effective drug target for intervening telomere function

A CLINICAL STUDY OF INHALANT ANAESTHESIA IN DOGS

A clinical trial was undertaken using three different inhalant anaesthetic agents and one intravenous anaesthetic agent in dogs undergoing routine desexing surgery. Healthy adult dogs undergoing either ovariohysterectomy or castration were assessed as to their demeanour, with the more excitable dogs being placed in groups receiving premedication with acepromazine and morphine. All dogs were then randomly assigned an anaesthetic agent for induction of general anaesthesia. The agents were the inhalants halothane, isoflurane and sevoflurane, and the intravenous agent propofol. Inhalant inductions were undertaken using a tight fitting mask attached to a standard anaesthetic machine with a rebreathing circuit, with the maximum dose of inhalant available from a standard vaporiser. Propofol inductions were undertaken via intravenous catheter. Dogs induced with propofol were randomly assigned one of the three inhalant agents for maintenance. Those induced by inhalant agent were maintained using the same agent. The surgical procedure was undertaken in standard fashion, as was recovery from anaesthesia. All dogs received the non-steroidal anti-inflammatory agent meloxicam. Data collection was divided into three stages: induction, maintenance, and recovery from anaesthesia. Variables measured at induction of anaesthesia were time to intubation, number of intubation attempts, tolerance of mask, quality of induction and quality of transfer to the maintenance stage. Standard variables for monitoring of anaesthesia were recorded throughout the maintenance of anaesthesia. Variables measured at recovery were time to righting, time to standing and quality of recovery. The mean time to intubation when using the newer inhalant sevoflurane (196.2 ± 14.8sec, mean ± SE) was not significantly different to that for halothane (221.4 ± 14.0sec) or isoflurane (172.4 ± 15.0sec). Time to intubation with isoflurane was significantly faster than with halothane. Mean time to intubation with propofol (85.4 ± 7.7sec) was significantly faster than that for any of the three inhalants. Choice of inhalant had no effect on quality of induction. The use of premedication significantly improved the quality of induction. The use of propofol for induction likewise significantly improved the quality of induction. Standard cardiorespiratory variables measured during the maintenance phase of anaesthesia remained within normal clinical ranges for all three inhalants, and were therefore not further analysed. Choice of inhalant agent had no significant effect on the time to righting or standing in recovery. The use of propofol for induction had no effect on these variables. Animals placed in groups receiving premedication had significantly longer times to righting and standing. The oesophageal temperature at the end of the procedure had a significant effect on times to righting and standing, with lower temperatures contributing to slower recoveries. Independent of procedure time, male dogs had shorter times to righting than female dogs

The influence of salinity on the effects of Multi-walled carbon nanotubes on polychaetes

Salinity shifts in estuarine and coastal areas are becoming a topic of concern and are one of the main factors influencing nanoparticles behaviour in the environment. For this reason, the impacts of multiwalled carbon nanotubes (MWCNTs) under different seawater salinity conditions were evaluated on the common ragworm Hediste diversicolor, a polychaete species widely used as bioindicator of estuarine environmental quality. An innovative method to assess the presence of MWCNT aggregates in the sediments was used for the first time. Biomarkers approach was used to evaluate the metabolic capacity, oxidative status and neurotoxicity of polychaetes after long-term exposure. The results revealed an alteration of energy-related responses in contaminated polychaetes under both salinity conditions, resulting in an increase of metabolism and expenditure of their energy reserves (lower glycogen and protein contents). Moreover, a concentration-dependent toxicity (higher lipid peroxidation, lower ratio between reduced and oxidized glutathione and activation of antioxidant defences and biotransformation mechanisms) was observed in H. diversicolor, especially when exposed to low salinity. Additionally, neurotoxicity was observed by inhibition of Cholinesterases activity in organisms exposed to MWCNTs at both salinities.publishe

The Telomere Binding Protein TRF2 Induces Chromatin Compaction

Mammalian telomeres are specialized chromatin structures that require the telomere binding protein, TRF2, for maintaining chromosome stability. In addition to its ability to modulate DNA repair activities, TRF2 also has direct effects on DNA structure and topology. Given that mammalian telomeric chromatin includes nucleosomes, we investigated the effect of this protein on chromatin structure. TRF2 bound to reconstituted telomeric nucleosomal fibers through both its basic N-terminus and its C-terminal DNA binding domain. Analytical agarose gel electrophoresis (AAGE) studies showed that TRF2 promoted the folding of nucleosomal arrays into more compact structures by neutralizing negative surface charge. A construct containing the N-terminal and TRFH domains together altered the charge and radius of nucleosomal arrays similarly to full-length TRF2 suggesting that TRF2-driven changes in global chromatin structure were largely due to these regions. However, the most compact chromatin structures were induced by the isolated basic N-terminal region, as judged by both AAGE and atomic force microscopy. Although the N-terminal region condensed nucleosomal array fibers, the TRFH domain, known to alter DNA topology, was required for stimulation of a strand invasion-like reaction with nucleosomal arrays. Optimal strand invasion also required the C-terminal DNA binding domain. Furthermore, the reaction was not stimulated on linear histone-free DNA. Our data suggest that nucleosomal chromatin has the ability to facilitate this activity of TRF2 which is thought to be involved in stabilizing looped telomere structures

Metal Bioavailability in the Sava River Water

Metals present one of the major contamination problems for freshwater systems, such as the Sava River, due to their high toxicity, persistence, and tendency to accumulate in sediment and living organisms. The comprehensive assessment of the metal bioavailability in the Sava River encompassed the analyses of dissolved and DGT-labile metal species of nine metals (Cd, Co, Cr, Cu, Fe, Mn, Ni, Pb, and Zn) in the river water, as well as the evaluation of the accumulation of five metals (Cd, Cu, Fe, Mn, and Zn) in three organs (liver, gills, and gastrointestinal tissue) of the bioindicator organism, fish species European chub (Squalius cephalus L.).This survey was conducted mainly during the year 2006, in two sampling campaigns, in April/May and September, as periods representative for chub spawning and post-spawning. Additionally, metal concentrations were determined in the intestinal parasites acanthocephalans, which are known for their high affinity for metal accumulation. Metallothionein concentrations were also determined in three chub organs, as a commonly applied biomarker of metal exposure. Based on the metal concentrations in the river water, the Sava River was defined as weakly contaminated and mainly comparable with unpolluted rivers, which enabled the analyses of physiological variability of metal and metallothionein concentrations in the chub organs, as well as the establishment of their constitutive levels