Influence of repeated applications of glyphosate on its persistence and soil bioactivity

Pesticide degradation studies are essential to evaluate its impact in the environment and on non-target organisms. The effect of repeated soil applications of the herbicide glyphosate on its dissipation and on soil microorganisms was studied by radiometric and microbial techniques. Results indicated fast dissipation of the [14C]-glyphosate or [14C]metabolites extractable residues (half-life of 0.92±0.29 month), but increasing half-lives of total mineralization ranging from 2.2 to 3.4 months as the number of applications increased from 1 to 4. No significant correlation was found between 14CO2 production and dehydrogenase activity.

Estudi de l'impacte ecotoxicològic de l'insecticida imidacloprid en diferents compartiments ambientals

En les darreres dècades la necessitat d’una major producció en l’agricultura ha implicat l’ús de productes químics per a millorar la producció. Entre aquests productes trobem els insecticides que, tot i ser específics per a determinades funcions, en molts casos acaben afectant també a altres organismes que no en són la diana. Els assajos d’ecotoxicitat són una eina clau per a determinar el grau d’afectació d’aquests insecticides. En aquest estudi es pretén determinar l’efecte de l’aplicació directa en el sòl de les dosis recomanades de l’insecticida Confidor 20SL (amb imidacloprid com a principi actiu) sobre Eisenia fetida i dels lixiviats d’aquests sòls sobre Daphnia magna i Selenastrum capricornutum simulant fenòmens d’escorrentia o lixiviació naturals. En el cas de E.fetida s’obté una LC50 de 24.71 mg/kg sòl i per la reproducció un valor de EC50 de 8.41 mg/kg sòl. S’observa allunyament en totes les dosis utilitzades i la EC50 és de 2.57 mg/kg sòl. No s’han pogut determinar efectes a nivell neurològic a partir de la determinació de l’activitat de l’ AChE. A l’exposar D.magna als lixiviats del sòl contaminat no s’han observat efectes clars ni en la mortalitat ni en la reproducció. El mateix succeeix amb S.capricornutum. Les dosis d’aplicació del pesticida Confidor, representen una amenaça per a E.fetida pel que fa a efectes subletals però no podem dir el mateix per als organismes aquàtics.

Influence of biocontrol strain Pseudomonas fluorescens CHA0 and its antibiotic overproducing derivative on the diversity of root colonizing pseudomonads

Non-target effects of biocontrol strains of Pseudomonas on the population of resident pseudomonads should be assessed prior to their large scale application in the environment. The rifampicin resistant bacterium P. fluorescens CHA0-Rif and its antibiotic overproducing derivative CHA0-Rif/pME3424 were introduced into soil microcosms and the population of resident pseudomonads colonizing cucumber roots was investigated after 10 and 52 days. Both CHA0-Rif and CHA0-Rif/pME3424 displaced a part of the resident pseudomonad population after 10 days. To investigate the population structure, utilization of 10 carbon sources and production of two exoenzymes was assessed for 5600 individual pseudomonad isolates and 1700 isolates were subjected to amplified ribosomal DNA restriction analysis of the spacer region (spacer-ARDRA). After 10 days, only the proportion of pseudomonads able to degrade -tryptophan was reduced in treatments inoculated with either biocontrol strain. In parallel the phenotypic diversity was reduced. These effects were only observed 10 days after inoculation, and they were similar for inoculation with CHA0-Rif and CHA0-Rif/pME3424. Changes in the population structure of resident pseudomonads on cucumber roots during plant growth were more pronounced than changes due to the inoculants. The inoculants did not affect the genotypic diversity detected with spacer-ARDRA, but the genotypic fingerprints corresponded only partially to the phenotypic profiles. Overall CHA0-Rif had a small and transient impact on the population of resident pseudomonads and the effect was essentially the same for the genetically engineered derivative CHA0-

The anticancer drug metabolites endoxifen and 4-hydroxy-tamoxifen induce toxic effects on Daphnia pulex in a two-generation study.

Although pharmaceutical metabolites are found in the aquatic environment, their toxicity on living organisms is poorly studied in general. Endoxifen and 4-hydroxy-tamoxifen (4OHTam) are two metabolites of the widely used anticancer drug tamoxifen for the prevention and treatment of breast cancers. Both metabolites have a high pharmacological potency in vertebrates, attributing prodrug characteristics to tamoxifen. Tamoxifen and its metabolites are body-excreted by patients, and the parent compound is found in sewage treatment plan effluents and natural waters. The toxicity of these potent metabolites on non-target aquatic species is unknown, which forces environmental risk assessors to predict their toxicity on aquatic species using knowledge on the parent compounds. Therefore, the aim of this study was to assess the sensitivity of two generations of the freshwater microcrustacean Daphnia pulex towards 4OHTam and endoxifen. Two chronic tests of 4OHTam and endoxifen were run in parallel and several endpoints were assessed. The results show that the metabolites 4OHTam and endoxifen induced reproductive and survival effects. For both metabolites, the sensitivity of D. pulex increased in the second generation. The intrinsic rate of natural increase (r) decreased with increasing 4OHTam and endoxifen concentrations. The No-Observed Effect Concentrations (NOECs) calculated for the reproduction of the second generation exposed to 4OHTam and endoxifen were <1.8 and 4.3μg/L, respectively, whereas the NOECs that were calculated for the intrinsic rate of natural increase were <1.8 and 0.4μg/L, respectively. Our study raises questions about prodrug and active metabolites in environmental toxicology assessments of pharmaceuticals. Our findings also emphasize the importance of performing long-term experiments and considering multi-endpoints instead of the standard reproduction outcome.

Imaging of Dopamine and Serotonin Transporters. Pre-clinical Studies with Radiotracers for Positron Emission Tomography

The action of the neurotransmitters dopamine (DA) and serotonin (5-HT) at synapses is terminated by their rapid reuptake into presynaptic nerve endings via plasma membrane dopamine (DAT) and serotonin (SERT) transporters. Alterations in the function of these transporters have been suggested as a feature of several neurological and neuropsychiatric diseases, such as Parkinson’s disease (PD), depression, and anxiety. A suitable clinical method for studying these transporters non-invasively in vivo is positron emission tomography (PET) utilizing radiopharmaceuticals (tracers) labelled with short-lived positron-emitting radionuclides. The aim of this study was to evaluate in rats two novel radiotracers, [¹⁸F]beta -CFT-FP and ¹⁸FFMe-McN, for imaging DAT and SERT, respectively, using in vitro, ex vivo and in vivo methods. Substituting an N-methyl in [¹⁸F]beta-CFT, a well known DAT tracer, with a ¹⁸Ffluoropropyl group significantly changed the properties of the tracer. [¹⁸F]beta- CFT showed slow kinetics and metabolism, and a high specific uptake in the striatum, whereas [¹⁸F]beta-CFT-FP showed fast kinetics and metabolism, and a moderate specific uptake in the striatum. [¹⁸F]betaCFT-FP was selective for DAT; but [¹⁸F]beta-CFT also bound to the noradrenaline transporter. [¹⁸F]beta-CFT-FP may be a suitable PET tracer for imaging the striatal DAT sites, but a tracer with a higher affinity is needed for imaging extrastriatal DAT sites. In rats, ¹⁸FFMe-McN showed high target-to-non-target ratios, specificity and selectivity for SERT, but slow kinetics. However, ¹⁸FFMe-McN reveals potential for imaging SERT, at least in pre-clinical studies. In addition, the sensitivities of [¹⁸F]beta CFT and [¹⁸ F]FDOPA (a precursor of DA) for detecting mild nigrostriatal hypofunction were compared in an animal model of PD. The uptake of [¹⁸F]FDOPA was significantly affected by compensatory effects in dopaminergic cells, whereas [¹⁸F]beta-CFT was more sensitive and therefore more suitable for PET studies of mild dopaminergic symptoms. In conclusion, both novel tracers, [¹⁸F]-CFT-FP and ¹⁸FFMe-McN, have potential, but are not optimal PET tracers for DAT and SERT imaging in rats, respectively. [¹⁸F]beta-CFT is superior to [18F]FDOPA for imaging mild nigral lesions in rat brains.

Characterisation of Cladonia rangiferina transcriptome and genome, and the effects of dehydration and rehydration on its gene expression

Lichens are symbiotic organisms, which consist of the fungal partner and the photosynthetic partner, which can be either an alga or a cyanobacterium. In some lichen species the symbiosis is tripartite, where the relationship includes both an alga and a cyanobacterium alongside the primary symbiont, fungus. The lichen symbiosis is an evolutionarily old adaptation to life on land and many extant fungal species have evolved from lichenised ancestors. Lichens inhabit a wide range of habitats and are capable of living in harsh environments and on nutrient poor substrates, such as bare rocks, often enduring frequent cycles of drying and wetting. Most lichen species are desiccation tolerant, and they can survive long periods of dehydration, but can rapidly resume photosynthesis upon rehydration. The molecular mechanisms behind lichen desiccation tolerance are still largely uncharacterised and little information is available for any lichen species at the genomic or transcriptomic level. The emergence of the high-throughput next generation sequencing (NGS) technologies and the subsequent decrease in the cost of sequencing new genomes and transcriptomes has enabled non-model organism research on the whole genome level. In this doctoral work the transcriptome and genome of the grey reindeer lichen, Cladonia rangiferina, were sequenced, de novo assembled and characterised using NGS and traditional expressed sequence tag (EST) technologies. RNA extraction methods were optimised to improve the yield and quality of RNA extracted from lichen tissue. The effects of rehydration and desiccation on C. rangiferina gene expression on whole transcriptome level were studied and the most differentially expressed genes were identified. The secondary metabolites present in C. rangiferina decreased the quality – integrity, optical characteristics and utility for sensitive molecular biological applications – of the extracted RNA requiring an optimised RNA extraction method for isolating sufficient quantities of high-quality RNA from lichen tissue in a time- and cost-efficient manner. The de novo assembly of the transcriptome of C. rangiferina was used to produce a set of contiguous unigene sequences that were used to investigate the biological functions and pathways active in a hydrated lichen thallus. The de novo assembly of the genome yielded an assembly containing mostly genes derived from the fungal partner. The assembly was of sufficient quality, in size similar to other lichen-forming fungal genomes and included most of the core eukaryotic genes. Differences in gene expression were detected in all studied stages of desiccation and rehydration, but the largest changes occurred during the early stages of rehydration. The most differentially expressed genes did not have any annotations, making them potentially lichen-specific genes, but several genes known to participate in environmental stress tolerance in other organisms were also identified as differentially expressed.

ERP and cardiovascular correlates of source memory discrimination in older and younger adults /

There is much evidence to support an age-related decline in source memory ability. However, the underlying mechanisms responsible for this decline are not well understood. The current study was carried out to determine the electrophysiological correlates of source memory discrimination in younger and older adults. Event-related potentials (ERPs) and continuous electrocardiographic (ECG) data were collected from younger (M= 21 years) and older (M= 71 years) adults during a source memory task. Older adults were more likely to make source memory errors for recently repeated, non-target words than were younger adults. Moreover, their ERP records for correct trials showed an increased amplitude in the late positive (LP) component (400-800 msec) for the most recently presented, non-target stimuli relative to the LP noted for target items. Younger adults showed an opposite pattern, with a large LP component for target items, and a much smaller LP component for the recently repeated non-target items. Computation of parasympathetic activity in the vagus nerve was performed on the ECG data (Porges, 1985). The resulting measure, vagal tone, was used as an index of physiological responsivity. The vagal tone index of physiological responsivity was negatively related to the LP amplitude for the most recently repeated, non-target words in both groups, after accounting for age effects. The ERP data support the hypothesis that the tendency to make source memory errors on the part of older adults is related to the ability to selectively control attentional processes during task performance. Furthermore, the relationship between vagal tone and ERP reactivity suggests that there is a physiological basis to the heightened reactivity measured in the LP response to recently repeated non-target items such that, under decreased physiological resources, there is an impairment in the ability to selectively inhibit bottom-up, stimulus based properties in favour of task-related goals in older adults. The inconsistency of these results with other explanatory models of source memory deficits is discussed. It is concluded that the data are consistent with a physiological reactivity model requiring inhibition of reactivity to irrelevant, but perceptually-fluent, stimuli.

Autonomic and electrocortical indices of performance monitoring and source memory discrimination in older and younger adults

Reduced capacity for executive cognitive function and for the autonomic control of cardiac responsivity are both concomitants of the aging process. These may be linked through their mutual dependence on medial prefrontal function, but the specifics ofthat linkage have not been well explored. Executive functions associated with medial prefrontal cortex involve various aspects ofperformance monitoring, whereas centrally mediated autonomic functions can be observed as heart rate variability (HRV), i.e., variability in the length of intervals between heart beats. The focus for this thesis was to examine the degree to which the capacity for phasic autonomic adjustments to heart rate relates to performance monitoring in younger and older adults, using measures of electrocortical and autonomic activity. Behavioural performance and attention allocation during two age-sensitive tasks could be predicted by various aspects of autonomic control. For young adults, greater influence of the parasympathetic system on HRV was beneficial for learning unfamiliar maze paths; for older adults, greater sympathetic influence was detrimental to these functions. Further, these relationships were primarily evoked when the task required the construction and use of internalized representations of mazes rather than passive responses to feedback. When memory for source was required, older adults made three times as many source errors as young adults. However, greater parasympathetic influence on HRV in the older group was conducive to avoiding source errors and to reduced electrocortical responses to irrelevant information. Higher sympathetic predominance, in contrast, was associated with higher rates of source error and greater electrocortical responses tq non-target information in both groups. These relations were not seen for 11 errors associated with a speeded perceptual task, irrespective of its difficulty level. Overall, autonomic modulation of cardiac activity was associated with higher levels of performance monitoring, but differentially across tasks and age groups. With respect to age, those older adults who had maintained higher levels of autonomic cardiac regulation appeared to have also maintained higher levels of executive control over task performance.

Age-related changes in ERPs associated with context-based and response-based interference

Age-related differences in information processing have often been explained through deficits in older adults' ability to ignore irrelevant stimuli and suppress inappropriate responses through inhibitory control processes. Functional imaging work on young adults by Nelson and colleagues (2003) has indicated that inferior frontal and anterior cingulate cortex playa key role in resolving interference effects during a delay-to-match memory task. Specifically, inferior frontal cortex appeared to be recruited under conditions of context interference while the anterior cingulate was associated with interference resolution at the stage of response selection. Related work has shown that specific neural activities related to interference resolution are not preserved in older adults, supporting the notion of age-related declines in inhibitory control (Jonides et aI., 2000, West et aI., 2004b). In this study the time course and nature of these inhibition-related processes were investigated in young and old adults using high-density ERPs collected during a modified Sternberg task. Participants were presented with four target letters followed by a probe that either did or did not match one of the target letters held in working memory. Inhibitory processes were evoked by manipulating the nature of cognitive conflict in a particular trial. Conflict in working memory was elicited through the presentation of a probe letter in immediately previous target sets. Response-based conflict was produced by presenting a negative probe that had just been viewed as a positive probe on the previous trial. Younger adults displayed a larger orienting response (P3a and P3b) to positive probes relative to a non-target baseline. Older adults produced the orienting P3a and 3 P3b waveforms but their responses did not differentiate between target and non-target stimuli. This age-related change in response to targetness is discussed in terms of "early selection/late correction" models of cognitive ageing. Younger adults also showed a sensitivity in their N450 response to different levels of interference. Source analysis of the N450 responses to the conflict trials of younger adults indicated an initial dipole in inferior frontal cortex and a subsequent dipole in anterior cingulate cortex, suggesting that inferior prefrontal regions may recruit the anterior cingulate to exert cognitive control functions. Individual older adults did show some evidence of an N450 response to conflict; however, this response was attenuated by a co-occurring positive deflection in the N450 time window. It is suggested that this positivity may reflect a form of compensatory activity in older adults to adapt to their decline in inhibitory control.

Analyse par spectrométrie de masse des antibiotiques vétérinaires liés à l’élevage porcin

L’élevage des porcs représente une source importante de déversement d’antibiotiques dans l’environnement par l’intermédiaire de l’épandage du lisier qui contient une grande quantité de ces molécules sur les champs agricoles. Il a été prouvé que ces molécules biologiquement actives peuvent avoir un impact toxique sur l’écosystème. Par ailleurs, elles sont aussi suspectées d’engendrer des problèmes sanitaires et de contribuer à la résistance bactérienne pouvant mener à des infections difficilement traitables chez les humains. Le contrôle de ces substances dans l’environnement est donc nécessaire. De nombreuses méthodes analytiques sont proposées dans la littérature scientifique pour recenser ces composés dans plusieurs types de matrice. Cependant, peu de ces méthodes permettent l’analyse de ces contaminants dans des matrices issues de l’élevage agricole intensif. Par ailleurs, les méthodes analytiques disponibles sont souvent sujettes à des faux positifs compte tenu de la complexité des matrices étudiées et du matériel utilisé et ne prennent souvent pas en compte les métabolites et produits de dégradation. Enfin, les niveaux d’analyse atteints avec ces méthodes ne sont parfois plus à jour étant donné l’évolution de la chimie analytique et de la spectrométrie de masse. Dans cette optique, de nouvelles méthodes d’analyses ont été développées pour rechercher et quantifier les antibiotiques dans des matrices dérivées de l’élevage intensif des porcs en essayant de proposer des approches alternatives sensibles, sélectives et robustes pour quantifier ces molécules. Une première méthode d’analyse basée sur une technique d’introduction d’échantillon alternative à l’aide d’une interface fonctionnant à l’aide d’une désorption thermique par diode laser munie d’une source à ionisation à pression atmosphérique, couplée à la spectrométrie de masse en tandem a été développée. L’objectif est de proposer une analyse plus rapide tout en atteignant des niveaux de concentration adaptés à la matrice étudiée. Cette technique d’analyse couplée à un traitement d’échantillon efficace a permis l’analyse de plusieurs antibiotiques vétérinaires de différentes classes dans des échantillons de lisier avec des temps d’analyse courts. Les limites de détection atteintes sont comprises entre 2,5 et 8,3 µg kg-1 et sont comparables avec celles pouvant être obtenues avec la chromatographie liquide dans une matrice similaire. En vue d’analyser simultanément une série de tétracyclines, une deuxième méthode d’analyse utilisant la chromatographie liquide couplée à la spectrométrie de masse à haute résolution (HRMS) a été proposée. L’utilisation de la HRMS a été motivée par le fait que cette technique d’analyse est moins sensible aux faux positifs que le triple quadripôle traditionnel. Des limites de détection comprises entre 1,5 et 3,6 µg kg-1 ont été atteintes dans des échantillons de lisier en utilisant un mode d’analyse par fragmentation. L’utilisation de méthodes de quantifications ciblées est une démarche intéressante lorsque la présence de contaminants est suspectée dans un échantillon. Toutefois, les contaminants non intégrés à cette méthode d’analyse ciblée ne peuvent être détectés même à de fortes concentrations. Dans ce contexte, une méthode d’analyse non ciblée a été développée pour la recherche de pharmaceutiques vétérinaires dans des effluents agricoles en utilisant la spectrométrie de masse à haute résolution et une cartouche SPE polymérique polyvalente. Cette méthode a permis l’identification d’antibiotiques et de pharmaceutiques couramment utilisés dans l’élevage porcin. La plupart des méthodes d’analyse disponibles dans la littérature se concentrent sur l’analyse des composés parents, mais pas sur les sous-produits de dégradation. L’approche utilisée dans la deuxième méthode d’analyse a donc été étendue et appliquée à d’autres classes d’antibiotiques pour mesurer les concentrations de plusieurs résidus d’antibiotiques dans les sols et les eaux de drainage d’un champ agricole expérimental. Les sols du champ renfermaient un mélange d’antibiotiques ainsi que leurs produits de dégradation relatifs à des concentrations mesurées jusqu’à 1020 µg kg-1. Une partie de ces composés ont voyagé par l’intermédiaire des eaux de drainage du champ ou des concentrations pouvant atteindre 3200 ng L-1 ont pu être relevées.

"|mpact of trawling on sea bottom and its living communities along the inshore waters of southwest coast of India'’

Mechanized fishing started in Indian waters in mid —fifties and large-scale operation of trawl fishing began in the mid sixties by the surfeit of individual entrepreneurs. The southwest coast of India especially the coastal waters of Kerala are the most productive area in the subcontinent and the state has been in the forefront in marine fish production (Kurup, 2001a). Though the coastline of Kerala is one tenth of the coastline of India, the state occupies the foremost position in the marine fish production of the country, accounting for more than 30% of the marine fish landings (Thomas, 2000). The coastal waters of Kerala have rich and diversified fishery resources, which are prone to heavy exploitation by a unprecedently high number of fishing gears, among them, mechanized bottom trawlers with a numerical strength of 4550 (Kurup, 2001a) against the permissible number of 1145 (Kalawar, et al., 1985) are the most destructive. Trawling operations during monsoon periods in Kerala has been a subject of controversy between traditional fishermen and trawl fishers on a subject that trawl fishing destroys large amount of juveniles and young ones of fishes since this period is the major breeding season of most of the fish and prawns (John, 1996). Therefore Government of Kerala imposed a ban on bottom trawling activities from 1988 onwards for a period varying from 21-70 days, which usually commences from June 15th. Though many studies revealed that large amount of non-target groups were destroyed in the commercial trawl fishing in the Indian waters, no concerted study has been conducted so far to evaluate the real impact of bottom trawling on the sea bottom and its living communities. The present study was conducted to assess the impact of excessive bottom trawling exerted on the sea bottom habitat and its living communities, which would be useful in impressing up on the seriousness of habitat degradation and biotic devastation, enabling the concerned to adopt relevant conservation and management steps to conserve the resources for sustainable exploitation

Soft Computing Approach for Optimization of siRNA Efficiency Prediction for Post-transcriptional Gene Silencing

Post-transcriptional gene silencing by RNA interference is mediated by small interfering RNA called siRNA. This gene silencing mechanism can be exploited therapeutically to a wide variety of disease-associated targets, especially in AIDS, neurodegenerative diseases, cholesterol and cancer on mice with the hope of extending these approaches to treat humans. Over the recent past, a significant amount of work has been undertaken to understand the gene silencing mediated by exogenous siRNA. The design of efficient exogenous siRNA sequences is challenging because of many issues related to siRNA. While designing efficient siRNA, target mRNAs must be selected such that their corresponding siRNAs are likely to be efficient against that target and unlikely to accidentally silence other transcripts due to sequence similarity. So before doing gene silencing by siRNAs, it is essential to analyze their off-target effects in addition to their inhibition efficiency against a particular target. Hence designing exogenous siRNA with good knock-down efficiency and target specificity is an area of concern to be addressed. Some methods have been developed already by considering both inhibition efficiency and off-target possibility of siRNA against agene. Out of these methods, only a few have achieved good inhibition efficiency, specificity and sensitivity. The main focus of this thesis is to develop computational methods to optimize the efficiency of siRNA in terms of “inhibition capacity and off-target possibility” against target mRNAs with improved efficacy, which may be useful in the area of gene silencing and drug design for tumor development. This study aims to investigate the currently available siRNA prediction approaches and to devise a better computational approach to tackle the problem of siRNA efficacy by inhibition capacity and off-target possibility. The strength and limitations of the available approaches are investigated and taken into consideration for making improved solution. Thus the approaches proposed in this study extend some of the good scoring previous state of the art techniques by incorporating machine learning and statistical approaches and thermodynamic features like whole stacking energy to improve the prediction accuracy, inhibition efficiency, sensitivity and specificity. Here, we propose one Support Vector Machine (SVM) model, and two Artificial Neural Network (ANN) models for siRNA efficiency prediction. In SVM model, the classification property is used to classify whether the siRNA is efficient or inefficient in silencing a target gene. The first ANNmodel, named siRNA Designer, is used for optimizing the inhibition efficiency of siRNA against target genes. The second ANN model, named Optimized siRNA Designer, OpsiD, produces efficient siRNAs with high inhibition efficiency to degrade target genes with improved sensitivity-specificity, and identifies the off-target knockdown possibility of siRNA against non-target genes. The models are trained and tested against a large data set of siRNA sequences. The validations are conducted using Pearson Correlation Coefficient, Mathews Correlation Coefficient, Receiver Operating Characteristic analysis, Accuracy of prediction, Sensitivity and Specificity. It is found that the approach, OpsiD, is capable of predicting the inhibition capacity of siRNA against a target mRNA with improved results over the state of the art techniques. Also we are able to understand the influence of whole stacking energy on efficiency of siRNA. The model is further improved by including the ability to identify the “off-target possibility” of predicted siRNA on non-target genes. Thus the proposed model, OpsiD, can predict optimized siRNA by considering both “inhibition efficiency on target genes and off-target possibility on non-target genes”, with improved inhibition efficiency, specificity and sensitivity. Since we have taken efforts to optimize the siRNA efficacy in terms of “inhibition efficiency and offtarget possibility”, we hope that the risk of “off-target effect” while doing gene silencing in various bioinformatics fields can be overcome to a great extent. These findings may provide new insights into cancer diagnosis, prognosis and therapy by gene silencing. The approach may be found useful for designing exogenous siRNA for therapeutic applications and gene silencing techniques in different areas of bioinformatics.

Contribution of antioxidant enzymes to toxicity assessment in fluvial biofilms

Per tal d’avaluar l’impacte de la contaminació en els ecosistemes aquàtics, aquesta tesi es centra en una aproximació multi-biomarcador en els biofilms. En complement dels biomarcadors clàssics, es va demostrar que les activitats dels enzims antioxidants (AEA): catalasa, ascorbat peroxidasa i glutatió reductasa eren biomarcardors d'estrès oxidatiu en els biofilms. Tot i que les AEA poden veure's influenciades amb la mateixa mesura per factors naturals (edat del biofilm, llum de colonització o d'exposició) i contaminants (herbicides i farmacèutics), aquestes AEA permeten entendre millor l'efecte dels contaminants. Cal remarcar que assajos de toxicitat aguda es poden utilitzar per comparar la capacitat antioxidant entre comunitats i conèixer la seva pre-exposició a l'estrès oxidatiu. Aquesta aproximació multi-biomarcador a nivell de comunitat és especialment interessant per avaluar la toxicitat dels contaminants emergents (β-blockers) sobre espècies no-diana. Per tal de millorar-la, també es va verificar la possibilitat de mesurar l'expressió gènica en biofilms.

Perception and modification of plant flavonoid signals by rhizosphere microorganisms

Flavonoids are a diverse class of polyphenolic compounds that are produced as a result of plant secondary metabolism. They are known to play a multifunctional role in rhizospheric plant-microbe and plant-plant communication. Most familiar is their function as a signal in initiation of the legume-rhizobia symbiosis, but, flavonoids may also be signals in the establishment of arbuscular mycorrhizal symbiosis and are known agents in plant defence and in allelopathic interactions. Flavonoid perception by, and impact on, their microbial targets (e.g. rhizobia, plant pathogens) is relatively well characterized. However, potential impacts on 'non-target' rhizosphere inhabitants ('non-target' is used to distinguish those microorganisms not conventionally known as targets) have not been thoroughly investigated. Thus, this review first summarizes the conventional roles of flavonoids as nod gene inducers, phytoalexins and allelochemicals before exploring questions concerning 'non-target' impacts. We hypothesize that flavonoids act to shape rhizosphere microbial community structure because they represent a potential source of carbon and toxicity and that they impact on rhizosphere function, for example, by accelerating the biodegradation of xenobiotics. We also examine the reverse question, 'how do rhizosphere microbial communities impact on flavonoid signals?' The presence of microorganisms undoubtedly influences the quality and quantity of flavonoids present in the rhizosphere, both through modification of root exudation patterns and microbial catabolism of exudates. Microbial alteration and attenuation of flavonoid signals may have ecological consequences for below-ground plant-microbe and plant-plant interaction. We have a lack of knowledge concerning the composition, concentration and bioavailability of flavonoids actually experienced by microbes in an intact rhizosphere, but this may be addressed through advances in microspectroscopic and biosensor techniques. Through the use of plant mutants defective in flavonoid biosynthesis, we may also start to address the question of the significance of flavonoids in shaping rhizosphere community structure and function.

Use of entomopathogenic bacterium Pseudomonas putida (Enterobacteriaceae) and it secretions against the Greater Wax Moth Galleria mellonella pupae

The bacterium from Pseudomonas putida from Steinernema abbasi and its metabolic secretions caused the mortality of the Galleria mellonella pupae. Experiments were conducted in sand and filter paper on time exposure, temperature, moisture, dose and time of penetration of bacterium in pupae and tested stored or dried toxic metabolites using G. mellonella pupae as a test target organism. Death of pupae was probably due to the toxic metabolites. Pseudomonas putida cells were recovered from the haemocoele when bacterial cells were applied to the G. mellonella pupae indicating that bacterial cells can enter the haemocoele in the absence of nematode vector. Penetration of bacterium was found rapidly after application on G. mellonella pupae. Pseudomonas putida or its toxic secretions can be used as a microbial control for insect control. The experimental results indicate that there is possibility of using P. putida and its toxic secretions as a biopesticide and can contribute in the development of new microbial and biological control against insect pests.