Muriel Raveton

Interactions between immune and biotransformation systems in fish: A review

In animals biotransformation and immune system are not totally independent, there are numerous functional interrelationships between these two systems. They are both implicated in the capacity of organisms to resist to a wide variety of environmental components such as viruses, bacteria and xenobiotics. It is known for a long time that the immune system functions as a physiologic system and interacts with all the other components of the organism including nervous or endocrine ones. In the same manner, the biotransformation system (especially the cytochrome P450 monooxygenases) is involved in the regulation of numerous hormone productions. In this way, many studies in mammals have revealed the possible interaction between immune and biotransformation systems. Among these interactions, the capacity of the activation of host defense mechanisms to down-regulate microsomal cytochrome P450 and the role of biotransformation system in the xenobiotic-mediated immunotoxicity have been underlined. Advances in the basic knowledge of fish immune and biotransformation systems should lead to a better understanding of the possible interactions between both systems and should improve fish health monitoring which is a crucial ecotoxicological goal.

A comparative study on the uptake and translocation of organochlorines by Phragmites australis

Organochlorines (OCs) are persistent chemicals found in various environmental compartments. The differences in the uptake of (14)C-labeled 1,4-dichlorobenzene (DCB), 1,2,4-trichlorobenzene (TCB) and γ-hexachlorocyclohexane (γHCH) by Phragmites australis were investigated under hydroponic conditions. The first step in sorption appears to be correlated with the hydrophobic nature of the compounds, since log-linear correlations were obtained between root concentration factor and partition coefficient (LogK(ow)). After 7 days of exposure, plant uptake of DCB, TCB, γHCH was significant with bioconcentration factors reaching 14, 19 and 15, respectively. Afterwards, uptake and translocation were seen to be more complex, with a loss of the simple relationship between uptake and LogK(ow). Linear correlations between the bioconcentration/translocation factors and the physico-chemical properties of OCs were shown, demonstrating that translocation from roots to shoots increases with solubility and volatility of the OCs. This suggests that OC-translocation inside plants might result from the combination of two processes, xylem sap flow and vapor fluxes. (14)C-phytovolatilization was measured and was correlated with the volatility of the compounds; the more volatile OCs being most the likely to be phytovolatilized from foliar surfaces (p=0.0008). Thus, OC-uptake/translocation appears to proceed at a rate that depends mostly on the OCs hydrophobicity, solubility and volatility.

Atrazine metabolism in corn seedlings

Atrazine is a herbicide widely used in corn culture. Corn seedlings metabolize this active ingredient readily in three ways. Two of them are catalysed by enzymes and another is purely chemical. In seedlings, the precise role played by each of these was still not clearly understood. Our work demonstrates that the chemical pathway leading to the formation of the inactive OH-2 atrazine is the pre-eminent form of metabolization inside the roots and, during the first week, inside the leaves. As shown by the kinetics of accumulation of the benzoxazinones responsible for this metabolization, a high potential of atrazine hydroxylation remains effective inside the leaves for at least 1 month. The OH-2 atrazine, which seems to accumulate inside the cell vacuoles, cannot move freely inside the plant and therefore cannot be abundantly transported from roots to leaves. The formation of a glutathione-atrazine conjugate, due to the activity of a glutathione-S-transferase (GST) isoenzyme, represents only a very small part of the total GST in corn. It is mostly present in the aerial parts and is highly effective only after a 1-week culture. When analysing the mixture of the metabolites formed in isolated corn leaves, which is very complex (nine products, among which six were major products), it seems that the metabolization pattern of corn leaves is the combined result of the three pathways and of the transformation of the conjugate into more simple derivatives.

Concentration responses to organochlorines in Phragmites australis.

Phragmites australis shows potential for the phytoremediation of chlorinated chemicals. Also there has been some attempt to determine the phytotoxic effects of organochlorines (OC). This study reports for lindane (HCH), monochlorobenzene (MCB), 1,4-dichlorobenzene (DCB) and 1,2,4-trichlorobenzene (TCB), a no-observed-effect-concentration (NOEC(7d)) that was 1000-300,000 times higher than environmental concentrations. Nevertheless, the combined OC mixture (NOEC(7d) level of each congener) induced a synergistic toxic effect, causing a severe drop (70%) in chlorophyll concentration. The mixture 0.2 mg L(-1) MCB+0.2 mg L(-1) DCB+2.5 mg L(-1) TCB+0.175 mg L(-1) HCH, that was 15 times more concentrated than environmental OC mixture, did not cause phytotoxicity during 21 days. Antioxidant enzymes were affected immediately after the start of exposure (3 days), but the plants showed no signs of stress thereafter. These data suggest that environmental OC mixtures do not pose a significant risk to P. australis.

Toxicokinetic of benzo[a]pyrene and fipronil in female green frogs (Pelophylax kl. esculentus)

A general consensus that an increased logK(ow) led to an increase in xenobiotic uptake and bioaccumulation is accepted. In this study we compared the toxicokinetics of two chemically different xenobiotics, i.e. benzo[a]pyrene and fipronil in female green frogs. Surprisingly, the uptake rates and the bioconcentration factors (BCF) of the two contaminants were not predicted by their logK(ow). The uptake rates obtained were of the same order of magnitude for the two contaminants and the BCFs measured for fipronil were about 3-fold higher than those obtained for benzo[a]pyrene. Fipronil appeared to be more recalcitrant than benzo[a]pyrene to detoxification processes leading to the accumulation of sulfone-fipronil especially in the ovaries. This phenomenon may explain reproductive influence of this contaminant described in other studies. Detoxification processes, including metabolism and the excretion of pollutants, are of importance when considering their persistence in aquatic organisms and trying to quantify their risks.

Kinetics of Uptake and Metabolism of Atrazine in Model Plant Systems

The present work concerns atrazine absorption and metabolism by corn (Zea mays.) seedlings immersed in an aqueous medium in comparison with Acer pseudoplatanus cell cultures. At the point of equilibrium, the apparent concentration inside the A. pseudoplatanus cells (with a moderate lipid content: 0.17% of dry weight) was about twice that of the medium. This equilibrium was probably due to a simple partition process; part of the atrazine was dissolved in the cell water and reached the same concentration as in the external medium while the rest was concentrated inside the cellular lipids. The theoretical calculation of the lipid/water partition, taking into account the value of log P measured not with the lipids but with octanol (log P = 2.5), gave a value of 1.5 for concentration inside the plant material. Such an equilibrium, resulting from a partition process between water and lipids, was also obtained in non-living corn seedlings. In living seedlings, an over-concentration of radioactivity due to [ 14 C]atrazine derivatives was rapidly obtained inside roots and shoots giving concentrations respectively 7- and 12-fold higher than that of atrazine in the external medium. This was due to very rapid chemical transformation of atrazine into its hydroxy derivatives, especially hydroxyatrazine. This hydrolysis of atrazine in corn was due to the presence of high levels of benzoxazinone derivatives in corn seedling cells. The hydroxylated metabolites were able to concentrate in the cells very rapidly and were unable to diffuse freely into the external medium. As a consequence, this process facilitated the penetration of large quantities of atrazine which became rapidly hydroxylated, allowing therefore the passive penetration of atrazine to be further improved, since the concentration C 1 in the receiver compartment was always close to zero. The passive transfer of atrazine, following Ficks law: dq/dt = - Pa (C 0 -C 1 ), was therefore optimized.

Impaired liver function in Xenopus tropicalis exposed to benzo[a]pyrene: transcriptomic and metabolic evidence

BackgroundDespite numerous studies suggesting that amphibians are highly sensitive to cumulative anthropogenic stresses, the role pollutants play in the decline of amphibian populations remains unclear. Amongst the most common aquatic contaminants, polycyclic aromatic hydrocarbons (PAHs) have been shown to induce several adverse effects on amphibian species in the larval stages. Conversely, adults exposed to high concentrations of the ubiquitous PAH, benzo[a]pyrene (BaP), tolerate the compound thanks to their highly efficient hepatic detoxification mechanisms. Due to this apparent lack of toxic effect on adults, no studies have examined in depth the potential toxicological impact of PAH on the physiology of adult amphibian livers. This study sheds light on the hepatic responses of Xenopus tropicalis when exposed to high environmentally relevant concentrations of BaP, by combining a high throughput transcriptomic approach (mRNA deep sequencing) and a characterization of cellular and physiological modifications to the amphibian liver.ResultsTranscriptomic changes observed in BaP-exposed Xenopus were further characterized using a time-dependent enrichment analysis, which revealed the pollutant-dependent gene regulation of important biochemical pathways, such as cholesterol biosynthesis, insulin signaling, adipocytokines signaling, glycolysis/gluconeogenesis and MAPK signaling. These results were substantiated at the physiological level with the detection of a pronounced metabolic disorder resulting in a possible insulin resistance-like syndrome phenotype. Hepatotoxicity induced by lipid and cholesterol metabolism impairments was clearly identified in BaP-exposed individuals.ConclusionsOur data suggested that BaP may disrupt overall liver physiology, and carbohydrate and cholesterol metabolism in particular, even after short-term exposure. These results are further discussed in terms of how this deregulation of liver physiology can lead to general metabolic impairment in amphibians chronically exposed to contaminants, thereby illustrating the role xenobiotics might play in the global decline in amphibian populations.

How human practices have affected vector-borne diseases in the past: a study of malaria transmission in Alpine valleys

BackgroundMalaria was endemic in the Rhône-Alpes area of eastern France in the 19th century and life expectancy was particularly shortened in Alpine valleys. This study was designed to determine how the disease affected people in the area and to identify the factors influencing malaria transmission.MethodsDemographic data of the 19th century were collected from death registers of eight villages of the flood-plain of the river Isère. Correlations were performed between these demographic data and reconstructed meteorological data. Archive documents from medical practitioners gave information on symptoms of ill people. Engineer reports provided information on the hydraulic project developments in the Isère valley.ResultsDescription of fevers was highly suggestive of endemic malaria transmission in the parishes neighbouring the river Isère. The current status of anopheline mosquitoes in the area supports this hypothesis. Mean temperature and precipitation were poorly correlated with demographic data, whereas the chronology of hydrological events correlated with fluctuations in death rates in the parishes.ConclusionNowadays, most of the river development projects involve the creation of wet areas, enabling controlled flooding events. Flood-flow risk and the re-emergence of vector-borne diseases would probably be influenced by the climate change. The message is not to forget that human disturbance of any functioning hydrosystem has often been linked to malaria transmission in the past.

The possible role of hydroxylation in the detoxification of atrazine in mature vetiver (Chrysopogon zizanioides nash) grown in hydroponics

The resistance mechanism of vetiver (Chrysopogon zizanioides) to atrazine was investigated to evaluate its potential for phytoremediation of environment contaminated with the herbicide. Plants known to metabolise atrazine rely on hydroxylation mediated by benzoxazinones, conjugation catalyzed by glutathione-S-transferases and dealkylation probably mediated by cytochromes P450. All three possibilities were explored in mature vetiver grown in hydroponics during this research project. Here we report on the chemical role of benzoxazinones in the transformation of atrazine. Fresh vetiver roots and leaves were cut to extract and study their content in benzoxazinones known to hydroxylate atrazine, such as 2,4-dihydroxy-2H-1,4-benzoxazin-3(4H)- one (DIBOA), 2,4-dihydroxy-7-methoxy-2H-1,4-benzoxazin-3(4H)-one (DIMBOA) and their mono- and di-glucosylated forms. Identification of benzoxazinones was performed by thin layer chromatography (TLC) and comparison of retention factors (Rf) and UV spectra with standards: although some products exhibited the same Rf as standards, UV spectra were different. Furthermore, in vitro hydroxylation of atrazine could not be detected in the presence of vetiver extracts. Finally, vetiver organs exposed to [14C]-atrazine did not produce any significant amount of hydroxylated products, such as hydroxyatrazine (HATR), hydroxydeethylatrazine (HDEA), and hydroxy-deisopropylatrazine (HDIA). Altogether, these metabolic features suggest that hydroxylation was not a major metabolic pathway of atrazine in vetiver.

Ubiquitous Water-Soluble Molecules in Aquatic Plant Exudates Determine Specific Insect Attraction

Plants produce semio-chemicals that directly influence insect attraction and/or repulsion. Generally, this attraction is closely associated with herbivory and has been studied mainly under atmospheric conditions. On the other hand, the relationship between aquatic plants and insects has been little studied. To determine whether the roots of aquatic macrophytes release attractive chemical mixtures into the water, we studied the behaviour of mosquito larvae using olfactory experiments with root exudates. After testing the attraction on Culex and Aedes mosquito larvae, we chose to work with Coquillettidia species, which have a complex behaviour in nature and need to be attached to plant roots in order to obtain oxygen. This relationship is non-destructive and can be described as commensal behaviour. Commonly found compounds seemed to be involved in insect attraction since root exudates from different plants were all attractive. Moreover, chemical analysis allowed us to identify a certain number of commonly found, highly water-soluble, low-molecular-weight compounds, several of which (glycerol, uracil, thymine, uridine, thymidine) were able to induce attraction when tested individually but at concentrations substantially higher than those found in nature. However, our principal findings demonstrated that these compounds appeared to act synergistically, since a mixture of these five compounds attracted larvae at natural concentrations (0.7 nM glycerol, <0.5 nM uracil, 0.6 nM thymine, 2.8 nM uridine, 86 nM thymidine), much lower than those found for each compound tested individually. These results provide strong evidence that a mixture of polyols (glycerol), pyrimidines (uracil, thymine), and nucleosides (uridine, thymidine) functions as an efficient attractive signal in nature for Coquillettidia larvae. We therefore show for the first time, that such commonly found compounds may play an important role in plant-insect relationships in aquatic eco-systems.