La neurotoxicité développementale associée au méthylmercure, au plomb et aux biphényles polychlorés : l’attention et le traitement visuel à l’étude

L’influence des contaminants environnementaux (CE) a été largement étudié en contexte d’exposition aigüe et d’intoxication. Qu’en est-il d’une exposition chronique à plus faibles doses sur le développement cognitif et sensoriel des enfants? Les études longitudinales de la présente thèse ont été réalisées au Nunavik. Bien que géographiquement éloignée des centres industriels, la communauté inuite est exposée aux CE via leur transport par les courants atmosphériques et océaniques ainsi que par leur bioaccumulation dans le poisson et les mammifères marins. Puisque l’alimentation autochtone traditionnelle, notamment la chair de béluga, est encore fréquemment consommée par la communauté, une proportion substantielle d’enfants inuits est exposée in utero aux CE. Ceux-ci sont également continuellement exposés à ces agents neurotoxiques durant leur développement postnatal. Or, la variation considérable dans l’adoption de l’alimentation traditionnelle au sein de la communauté représente une opportunité pour étudier la relation entre les niveaux d’exposition aux CE et le développement cognitif et sensoriel des enfants. Bien que certains déficits aient déjà été mis en lien avec l’exposition chronique aux CE dans la littérature, la présente thèse s’intéressa à cette relation plus spécifiquement chez les enfants inuits vivant dans le Grand Nord et plus exposés aux CE en raison de leur alimentation. Par ailleurs, les protocoles qui ont été développés pour cette thèse permettront d’évaluer des aspects qui ont été peu étudiés en lien avec les CE, soit l’attention visuospatiale et le traitement visuel cérébral précoce. Dans le premier volet de cette thèse, la relation entre trois CE et l’attention visuospatiale a été étudiée à l’aide d’une version adaptée de la tâche de Posner (M.I. Posner et al., 1980). Cette tâche psychophysique a été administrée à des enfants inuits (âge moyen = 11.2 ans) dont les niveaux d’exposition au mercure (Hg), au plomb (Pb) et aux biphényles polychlorés (BPCs) ont été documentés durant les périodes pré et postnatale. Les expositions in utero au Pb et aux BPCs ont été significativement associées à de l’impulsivité et de l’inattention, respectivement, alors que l’exposition postnatale au Pb a été associée à des temps de réaction plus longs. Bien qu’aucune relation spécifique avec l’attention visuospatiale n’ait été trouvée, les résultats de cette étude suggèrent que l’exposition aux CE est associée à une diminution des capacités attentionnelles générales chez les enfants résidant au Nunavik. Dans le second volet, le traitement cérébral précoce de l’information visuelle a été examiné à l’aide de potentiels évoqués visuels auprès d’enfants de la même communauté (âge moyen = 10.9 ans). La concentration de Hg dans le sang de cordon ombilical mesurée à la naissance a été associée à une réduction de l’amplitude et à une augmentation de la latence de la composante N75. L’exposition prénatale au Pb a quant à elle été associée à un délai dans la latence de la composante N150. Les résultats obtenus suggèrent ainsi que l’exposition aux métaux lourds, en particulier durant la période gestationnelle, serait associée à des altérations dans le développement visuel. Les résultats présentés dans cette thèse soutiennent l’hypothèse selon laquelle le cerveau en développement est vulnérable lors d’une exposition chronique aux CE, et cela même à des niveaux de concentration inférieurs aux limites recommandées par les organismes de santé publique. Les résultats permettent également d’apporter un éclairage nouveau sur les déficits cognitifs et sensoriels associés aux CE.

Nouvelles stratégies pour l’analyse des cyanotoxines par spectrométrie de masse

Les cyanobactéries ont une place très importante dans les écosystèmes aquatiques et un nombre important d’espèces considéré comme nuisible de par leur production de métabolites toxiques. Ces cyanotoxines possèdent des propriétés très variées et ont souvent été associées à des épisodes d’empoisonnement. L’augmentation des épisodes d’efflorescence d’origine cyanobactériennes et le potentiel qu’ils augmentent avec les changements climatiques a renchéri l’intérêt de l’étude des cyanobactéries et de leurs toxines. Considérant la complexité chimique des cyanotoxines, le développement de méthodes de détection simples, sensibles et rapides est toujours considéré comme étant un défi analytique. Considérant ces défis, le développement de nouvelles approches analytiques pour la détection de cyanotoxines dans l’eau et les poissons ayant été contaminés par des efflorescences cyanobactériennes nuisibles a été proposé. Une première approche consiste en l’utilisation d’une extraction sur phase solide en ligne couplée à une chromatographie liquide et à une détection en spectrométrie de masse en tandem (SPE-LC-MS/MS) permettant l’analyse de six analogues de microcystines (MC), de l’anatoxine (ANA-a) et de la cylindrospermopsine (CYN). La méthode permet une analyse simple et rapide et ainsi que la séparation chromatographique d’ANA-a et de son interférence isobare, la phénylalanine. Les limites de détection obtenues se trouvaient entre 0,01 et 0,02 μg L-1 et des concentrations retrouvées dans des eaux de lacs du Québec se trouvaient entre 0,024 et 36 μg L-1. Une deuxième méthode a permis l’analyse du b-N-méthylamino-L-alanine (BMAA), d’ANA-a, de CYN et de la saxitoxine (STX) dans les eaux de lac contaminés. L’analyse de deux isomères de conformation du BMAA a été effectuée afin d’améliorer la sélectivité de la détection. L’utilisation d’une SPE manuelle permet la purification et préconcentration des échantillons et une dérivatisation à base de chlorure de dansyle permet une chromatographie simplifiée. L’analyse effectuée par LC couplée à la spectrométrie de masse à haute résolution (HRMS) et des limites de détections ont été obtenues entre 0,007 et 0,01 µg L-1. Des échantillons réels ont été analysés avec des concentrations entre 0,01 et 0,3 µg L-1 permettant ainsi la confirmation de la présence du BMAA dans les efflorescences de cyanobactéries au Québec. Un deuxième volet du projet consiste en l’utilisation d’une technologie d’introduction d’échantillon permettant des analyses ultra-rapides (< 15 secondes/échantillons) sans étape chromatographique, la désorption thermique à diode laser (LDTD) couplée à l’ionisation chimique à pression atmosphérique (APCI) et à la spectrométrie de masse (MS). Un premier projet consiste en l’analyse des MC totales par l’intermédiaire d’une oxydation de Lemieux permettant un bris de la molécule et obtenant une fraction commune aux multiples congénères existants des MC. Cette fraction, le MMPB, est analysée, après une extraction liquide-liquide, par LDTD-APCI-MS/MS. Une limite de détection de 0,2 µg L-1 a été obtenue et des concentrations entre 1 et 425 µg L-1 ont été trouvées dans des échantillons d’eau de lac contaminés du Québec. De plus, une analyse en parallèle avec des étalons pour divers congénères des MC a permis de suggérer la possible présence de congénères ou d’isomères non détectés. Un deuxième projet consiste en l’analyse directe d’ANA-a par LDTD-APCI-HRMS pour résoudre son interférence isobare, la phénylalanine, grâce à la détection à haute résolution. La LDTD n’offre pas de séparation chromatographique et l’utilisation de la HRMS permet de distinguer les signaux d’ANA-a de ceux de la phénylalanine. Une limite de détection de 0,2 µg L-1 a été obtenue et la méthode a été appliquée sur des échantillons réels d’eau avec un échantillon positif en ANA-a avec une concentration de 0,21 µg L-1. Finalement, à l’aide de la LDTD-APCI-HRMS, l’analyse des MC totales a été adaptée pour la chair de poisson afin de déterminer la fraction libre et liée des MC et comparer les résultats avec des analyses conventionnelles. L’utilisation d’une digestion par hydroxyde de sodium précédant l’oxydation de Lemieux suivi d’une purification par SPE a permis d’obtenir une limite de détection de 2,7 µg kg-1. Des échantillons de poissons contaminés ont été analysés, on a retrouvé des concentrations en MC totales de 2,9 et 13,2 µg kg-1 comparativement aux analyses usuelles qui avaient démontré un seul échantillon positif à 2 µg kg-1, indiquant la possible présence de MC non détectés en utilisant les méthodes conventionnelles.

Standardization of Optimum Conditions for the Production of Glucosamine Hydrochiloride from Chitin

The study entitled standardization of optimum conditions for the production of glucosamine hydrochloride from chitin. Shellfish processing industries around the world turn out a significant quantity of head and shell as industrial waste. The waste must be removed immediately to prevent the contamination to the processing environment. The technique that are available for their disposal include ocean dumping, incineration or disposal of landfill sites. In this thesis the techniques and methods are used to process glucosamine hydrochloride from crustacean processing waste. Chitin is a nitrogenous polysaccharide, which is white, hard, inelastic, found in outer skeleton of insects, crabs, shrimp and lobsters and in the internal structures of other invertebrates. Glucosamine can be considered as a nutraceutical product by virtue of its properties. It is important for healthy skin, and plays a major role in the healing of surgical incisions and skin wounds. Deproteinisation of shrimp shell had significant effect on quality of chitin. Demineralization is also influences chitin quality. Solvents used for glucosamine hydrochloride affects the final yield and purity.

STUDIES ON TAXONOMY, DISTRIBUTION, ECOLOGY AND REPRODUCTIVE POTENTIAL OF ROTIFERS FROM SELECTED CENTRES IN COCHIN BACKWATER SYSTEM, KERALA

The studies were conducted in nine stations with varying ecological characteristics along Cochin backwaters and adjoining canals. Many workers opined that the distribution of rotifers is cosmopolitan. The significance of rotifers as first food for early larvae was indicated by Fujita. Aquaculture is a fast growing field in fisheries sector and it is gaining more importance as the fish landings and supply are getting irregular. A consistent supply of fish/shellfish can only be achieved through aquaculture. The success of any culture activity depends on the timely production of seeds of finfishes/shellfishes. The availability of wild seed is seasonal and erratic. So, a dependable source of seed of fishes and shellfishes is possible only through large scale production in hatchery. A successful seed production activity depends on the availability of a variety of suitable live feed organisms in sufficient quantities at the proper time for use in the larval stages. As the live feeds promote high growth rates, easy digestion, assimilation and the quality of not contaminating the culture water when compared to other artificial feeds, make the culture of live feed organisms the principal means of providing food for the larvae of finfishes and shellfishes. Rotifers are considered to be an excellent and indispensable food for larvae of many finfishes and crustaceans. It (1960) was the first to culture Brachionus plicatilis for feeding marine fish larvae, and now it is being extensively used as live feed in hatcheries all over the world. They are a group of microscopic organisms coming under the Phylum Rotifera which comprises of about 2000 species. Their slow swimming habits, ability to tolerate a wide range of salinities, parthenogenetic mode of reproduction and ability to get enriched easily, make rotifers an ideal live feed organism. The major factors such as temperature, salinity and food that influence the reproductive potential and thereby the population size of rotifer, Salinity is one of the most important aspect influencing the reproductive rate of rotifers. The feed type and feed concentration play a vital role in influencing the reproductive rate of rotifers. For culture of rotifers, the commonly used micro algae belong to Chlorella, Nannochloropsis, Isochrysis and Tetraselmis. While some studies have suggested that, algal diet has little effect on reproductive rates in 1979 while using the rotifer, Brachionus plicatilis as feed for the larvae of red sea bream, Pagrus major. It is generally accepted that rotifers play a pivotal role in the successful rearing of marine fish larvae.

Studies on the prevalence of algal blooms along Kerala coast, India

Algal blooms are naturally occurring phenomena in the aquatic environment. These blooms cause mass mortalities of wild and farmed fish and shellfish, human intoxications which sometimes result in death, alteration of marine trophic structure through adverse effects on larvae and other life history stages of commercially important species and death of marine animals. Occurrences of harmful algal blooms and associated mortality have been reported along the coastal waters of India since the early period of the last century. The present study was taken up to study the dynamics of major phytoplankton blooms, which occur along the Kerala coast. The results of quantitative and qualitative analysis of phytoplankton in the coastal waters of Vizhinjam and Chombala, their species diversity and community structure is presented and the major algal blooms recorded along the coast of Kerala during the study period is described and their occurrence is related to the hydrographic and meteorological variations. There is a clear evident from these works in the Indian region that the fishes avoid areas where these harmful algae bloom, either due to the toxicity or due to some irritant property of the chemicals secreted by the algae. Taxonomic diversity studies indicated a change in the community structure of commercial finfishes, crustaceans and molluscs due to the bloom of C.marina and funnel plots indicated the deviation in taxonomic distinctness during the bloom period from theoretical mean for the region.

lncidence of enterotoxigenic Staphylococcus aureus in relation to the microbial safety of seafood

A detailed study was made on the microbial quality, with special reference to food safety, of the fish and fishery products in the retail trade in Cochin and around. Also, farmed molluscan shellfishes like mussels and oysters were investigated for the microbial quality including the presence of pathogenic bacteria. Special stress has been given to monitor the incidence of coagulase positive as well as coagulase negative Staphylococcus in these products and their relative incidence have been recorded.In the next part, the investigation was centered mainly on toxigenic S.aureus. This is because among the Gram positive toxigenic bacteria, the Saureus with potential to produce thermostable enterotoxins are more relavent in food safety conceming seafoods in comparison with the Gram-negative pathogens like Salmonella and V.cholerae.The incidence, toxigenic potential and conditions of toxin production by S.aureus have been investigated in detail. An attempt has also been made to relate the toxigenisis with the presence of the concerned toxigenic genes in the genomes of S. aureus strains.

Effect of some toxic metals on selected phytoplankton of Kerala water

The extraction and use of metals has been the mainstay for the sustained development and progress of a nation. Metals, though fairly stable in the natural environment are found in trace quantities in water bodies. Attention has therefore been focused to identify the metals that impair the water quality. In the last few decades the concern about the fate of these metals in the aquatic system has been gaining momentum, particularly in the industrial belts. The disasters caused by metal poisoning in recent times have prompted an indepth study of the interaction of metals with aquatic biota. Kerala, basically an agriculture oriented state has witnessed the upsurgence of various industries as a part of the nationwide economic development programme. Cochin has been identified as the industrial capital of the state.The present study is an attempt towards a better understanding of the metal-phytoplankton interactions with special reference to the physiological changes in the species. various parameters such as temperature, salinity, pH, nutrients, number of cells, photosynthetic pigments, carbohydrates, protein and lipid are studied to highlight the complexity of metal..phytoplankton interaction

Chitinase production by.marine fungi

In this thesis an attempt is made to explore the potential of marine fungi for the production of chitinolytic enzymes and to recognize the ability to hydrolyse native chitin through submerged as well as solid substrate fermentation culture conditions, using wheat bran and shellfish processing waste such as ‘prawn waste’ as solid substrates. Attempt was made to isolate a potential chitinase producing fungus from marine environment and to develop an ideal bioprocess for the production ofchitolytic enzymes.Present study indicate scope for utilization of B. bassiana for industrial production of chitinase using prawn waste as solid substrate employing solid substrate fermentation.

Influence of collagen content on the texture andstorage quality of fish mince basedproducts

Objectives of the present study are to find out the proximate composition of 20 commercially important tropical fish species on the west coast of India. To determine the collagen content in these commercially important fish species and fractionation of collagen into acid soluble collagen (ASC) and hot water soluble (insoluble) collagen (ISC). To classify fishes according to its collagen content and To study the different storage characteristics in the mince based product—surimi, from different species of fishes. The researcher tries to find out a suitable collagen source to incorporate in surimi. and studies the different storage qualities in the mince based product, surimi at different levels of collagen in different species of fishes. The optimum collagen level to get desirable texture and storage quality for mince based product. The researcher aims to develop some products from surimi with desirable level of collagen. And compare the products prepared from surimi of lesser collagen content fish containing desirable level of collagen with surimi prepared with high collagen content fish without collagen. This study gains in importance as there is littleinformation on the collagen content of different species of fishes in India. So far no attempt was made to classify fishes according to its collagen content.

Nutritional Requirements Of The Fry Of Gold-Spot Mullet Liza Parsa (Hamilton)

Most tropical aquatic environments are naturally fertile and their natural fertility is renewed very rapidly. Natural food for many cultivable organisms can be grown to the maximum by proper management. However, enrichment of the environment can be done through rational fertilization. Still further increase in stocking rates, can yield increased crop if adequate feeding is done. Thus fish and shellfish nutrition is an important aspect of the multidisciplinary subject of aquaculture. The oldest and most classical studies in physiology have investigated the nutritional needs of the species of interest to aquaculture. The alimentary requirements for proteins, lipids, mineral salts and vitamins have been established for some temperate species. But, the nutritional requirements of only few tropical species have been studied. Before formulating a diet, a thorough knowledge of the nutrient requirement of the species is essential. It is against this background that the present area of investigation has been identified. "Nutritional requirements of the fry of gold-spot mullet Liza parsia" is a comprehensive attempt to quantify the nutritional factors that are essential for producing healthy fingerlings for stocking the farms. Aspects such as the protein and lipid requirements of the fry, the vitamin essentiality, nutritive evaluation of protein and lipid sources suitable for compounding diets were covered in this research project. The ultimate aim has been to evolve practical diets which could be applied in the nursery phase for juvenile production.

Chemical Evaluation of Selected Organics and Trace Metals in the Mangrove Macroflora Of Cochin

Mangroves are considered to play a significant role in global carbon cycling. Themangrove forests would fix CO2 by photosynthesis into mangrove lumber and thus decrease the possibility of a catastrophic series of events - global warming by atmospheric CO2, melting of the polar ice caps, and inundation of the great coastal cities of the world. The leaf litter and roots are the main contributors to mangrove sediments, though algal production and allochthonous detritus can also be trapped (Kristensen et al, 2008) by mangroves due to their high organic matter content and reducing nature are excellent metal retainers. Environmental pollution due to metals is of major concern. This is due to the basic fact that metals are not biodegradable or perishable the way most organic pollutants are. While most organic toxicants can be destroyed by combustion and converted into compounds such as C0, C02, SOX, NOX, metals can't be destroyed. At the most the valance and physical form of metals may change. Concentration of metals present naturally in air, water and soil is very low. Metals released into the environment through anthropogenic activities such as burning of fossils fuels, discharge of industrial effluents, mining, dumping of sewage etc leads to the development of higher than tolerable or toxic levels of metals in the environment leading to metal pollution. Of course, a large number of heavy metals such as Fe, Mn, Cu, Ni, Zn, Co, Cr, Mo, and V are essential to plants and animals and deficiency of these metals may lead to diseases, but at higher levels, it would lead to metal toxicity. Almost all industrial processes and urban activities involve release of at least trace quantities of half a dozen metals in different forms. Heavy metal pollution in the environment can remain dormant for a long time and surface with a vengeance. Once an area gets toxified with metals, it is almost impossible to detoxify it. The symptoms of metal toxicity are often quite similar to the symptoms of other common diseases such as respiratory problems, digestive disorders, skin diseases, hypertension, diabetes, jaundice etc making it all the more difficult to diagnose metal poisoning. For example the Minamata disease caused by mercury pollution in addition to affecting the nervous system can disturb liver function and cause diabetes and hypertension. The damage caused by heavy metals does not end up with the affected person. The harmful effects can be transferred to the person's progenies. Ironically heavy metal pollution is a direct offshoot of our increasing ability to mass produce metals and use them in all spheres of existence. Along with conventional physico- chemical methods, biosystem approachment is also being constantly used for combating metal pollution

Studies on The Eco Biology of The Indian Pearl Oyster, Pinctada Fucata (Gould)

The present work aims to study induced maturation of the pearl oyster for induced spawning experiments. The work on larval development was done with a view to developing techniques for the artificial rearing of commercially important pearl oyster P fucata, and also to elucidate the principles and problems of tropical bivalve larvae in general for detailed investigations in the future. The present study is designed to probe into the details of the basic aspects of the biology related to the hatchery technology of Pinctada fucata and the understanding of the factors which influence induction of maturation, spawning, larval rearing and spat settlement. This would go a long way in the upgradation of hatchery technology of the Indian Pearl oyster Pinctada fucata fora commercial level seed production..

“Stress Responses of Stinging Catfish Heteropneustes fassilis (Bloch) to Organophosphorus Insecticide Monocrotophos”

Poisoning by pesticides from agricultural fields is a serious water pollution problem and its environmental long-term effect may result in the incidence of poisoning of fish and other aquatic life forms (jyothi and Narayan, 1999). Fishes like Heteropneustesfbssilis and C/arius batrac/nus are especially prone to serious pesticide pollution as their habitat is mostly the agriculture area. Though only few studies are conducted in this area, it can be assessed from the local information that, population of such fish is on the verge of vulnerability due to extensive use of pesticides. The knowledge of sublethal effects of xenobiotic compounds on hematological parameters, enzyme activities and metabolite concentrations is very important to delineate the fish health status and provide a future understanding of ecological impacts. These pesticides act by causing inhibition of cholinesterase enzymes (ChE) by formation of enzyme inhibitor complex (O'Brien, 1976) and damaging the nervous system. These effects may result in metabolic disorders. Associated to cholinesterase activities, a study of other enzymes such as phosphatases and aminotransferases close to intermediary metabolite determination provides a wider view of metabolism. Interest in toxicological aspects has grown in recent years and research is now increasingly focused on mechanistic aspects of oxidative damage and cellular responses in biological system. The term ‘biomarker’ is generally used in a broad sense to include almost any measurement reflecting an interaction between a biological system and a potential hazard, which may be chemical, physical or biological (WHO, 1993). As biomarker stands for immediate responses, they are used as early warning signals of biological effects caused by environmental pollutants. The present work attempts to assess the toxicity of organophosphorus insecticide monocrotophos on the experimental organism selected for this study namely stinging catfish (Heteropneustesfossi/is) (Bloch), and to probe into the stress responses of the organism

Molecular Approaches For Characterization And Determination Of Pathogenicity Of Vibrios With Special Reference To Vibrio Harveyi From Penaeus Monodon Larval Production Systems

The present study focuses on vibrios especially Vibrio harveyi isolated from shrimp (P. monodon) larval production systems from both east and west coasts during times of mortality. A comprehensive approach has been made to work out their systematics through numerical taxonomy and group them based on RAPD profiling and to segregate the virulent from non- virulent isolates based on the presence of virulent genes as well as their phenotypic expression. The information gathered has helped to develop a simple scheme of identification based on phenotypic characters and segregate the virulent from non virulent strains of V. harveyi.