The study of marine antifreeze proteins has provided new findings. The blue mussel (Mytilus edulis) was known to have antifreeze activity; however, the antifreeze protein or other molecule responsible has never been characterized. Activity was evident in mussels from each of the Maritime provinces, Canada. The antifreeze molecule was shown to alter ice crystal morphology. It functioned over a wide range of pH values and it showed protease resistance. Nonetheless, its purification was not achieved. A winter flounder (Pseudopleuronectes americanus) ?-helical antifreeze protein, wflAFP6, has been shown to form amyloid-like fibrils during freezing. Separation of different aspects of the freezing process demonstrated that equilibrium freezing with an ice template is necessary for conversion of the wflAFP6 to the amyloid-like conformation. Amyloid-like conformation was determined by dye binding and electron microscopy. The effects of wflAFP6 concentration and solution properties were determined in order to better understand the process of conversion.

The current explanation for the absence, or low abundance, of filter-feeding invertebrates from some rocky shores is that because of local variation in nearshore oceanographic conditions, larvae do not arrive in sufficient numbers to establish populations. One putative consequence of this is that macroalgae are able to establish dominance in areas where filter-feeders (especially mussels) do not recruit well. While macroalgae have been transplanted to mussel-dominated shores with varying success, the survival, growth and reproduction of transplanted mussels has not been tested in areas dominated by macroalgae. To determine specifically what tips the balance between shores dominated by filter-feeding invertebrates and those dominated by macroalgae, I monitored the recruitment of intertidal mussels at four sites on the Kaikoura coast: two with mussels present and two algal-dominated. No significant differences in mussel recruitment rates were found between habitats and recruitment intensity at all sites was found to be very low. Recruitment limitation is not the reason for the absence of mussels from algal dominated shores but some form of limitation does occur to reduce the number of arriving mussels. Predation effects were examined by transplanting juvenile mussels into caged, uncaged and control treatments. No significant differences in predation rates between habitats were found and transplanted mussels in open cages at all sites were removed within 3 days. Mobile fish predators appeared to be the most likely cause of this intense predation. Growth of transplanted mussels into algal and mussel habitats was found to be significantly different. Mussels grew faster in mussel dominated habitats and after 6 months in algal dominated habitats, all mussels had died. The outcome of these experiments indicates that there is a close relationship between recruitment, survival and growth which tips the balance and allows the existence of mussel beds along the Kaikoura coastline.

Dissertação (mestrado) - Universidade Federal de Santa Catarina, Centro de Ciências Biológicas, Programa de Pós-Graduação em Bioquímica, Florianópolis, 2010 / Made available in DSpace on 2012-10-25T07:35:59Z (GMT). No. of bitstreams: 1 284569.pdf: 1495105 bytes, checksum: b4504c0a28e414d6b42d9ecd0498ac9b (MD5) / As espécies reativas de oxigênio (ERO), metabólitos secundários do metabolismo do oxigênio, são moléculas geradas e reguladas constantemente pela célula. O aumento na sua produção pode levar a inúmeros processos patológicos, e para evitar esses efeitos tóxicos das ERO a célula conta com um complexo sistema de defesas celulares, incluindo defesas antioxidantes. Esse sistema é bem estudado e caracterizado em eucariotos mais complexos, como os mamíferos, e em eucariotos mais simples, como as leveduras, mas ainda pouco se sabe sobre esse sistema e sua regulação em invertebrados marinhos. Desta forma, o presente trabalho tem como objetivo estudar o sistema antioxidante de bivalves marinhos em situação de estresse oxidativo, e propor possíveis mecanismos de modulação e regulação redox nesses modelos animais. O trabalho foi dividido em dois experimentos: no primeiro, mexilhões azuis Mytilus edulis foram pré-expostos ao selênio (4µg/L) por 3 dias, e subseqüentemente expostos por mais 3 dias a cobre (56 µg/L). Neste experimento, objetivamos detectar se os efeitos oxidativos do cobre podem ser atenuados pela pré-exposição ao selênio. Vários parâmetros antioxidantes e de dano celular foram avaliados. Nesta primeira etapa, observou-se que o cobre causa oxidação de tióis protéicos (PSH) e inibição da enzima tioredoxina redutase (TrxR) na brânquia e induz danos no DNA em hemócitos. O selênio aumenta a atividade da enzima glutationa peroxidase dependente de selênio (GPx-Se) e os níveis de glutationa (GSH). Quando os animais são pré-expostos a selênio e expostos em seguida ao cobre, a pré-exposição ao selênio é capaz de neutralizar os efeitos deletérios desse metal, revertendo a oxidação dos PSH, evitando a indução de danos no DNA e revertendo parcialmente a inibição da TrxR. No segundo experimento, mexilhões Perna perna foram expostos por até 21 dias ao zinco (10 µM), um metal conhecido por afetar o metabolismo da glutationa. A exposição aguda (2 dias) ao zinco causou consumo de GSH e inibição da GR, porém estes valores retornam aos níveis basais após 7 e 21 dias de exposição. Os animais expostos por 7 e 21 dias ao zinco demonstram uma amplificação do sistema antioxidante, com aumento na atividade de diversas enzimas auxiliares (TrxR, GR) ou de detoxificação de ERO (catalase, glutationa peroxidase total, superóxido dismutase). Apesar disso, essa amplificação não impediu um aumento nos níveis de peróxidos citosólicos e nos índices de peroxidação lipídica evidenciados após a exposição por 7 ou 21 dias. O conjunto de dados desses dois experimentos demonstram a importância do sistema antioxidante para esses animais em situações de estresse oxidativo. O aumento dos níveis de GSH e da atividade da GPx-Se parecem ser uma importante via de proteção contra os danos oxidativos em mexilhões M. edulis, mediados pelo cobre. Já com relação ao zinco, o restabelecimento do metabolismo da glutationa, o qual é afetado pelo zinco, é fundamental para as células branquiais de mexilhões P. perna, o qual responde ao insulto oxidativo com uma forte resposta adaptativa de amplificação das defesas antioxidantes celulares.

The Mediterranean mussel Mytilus galloprovincialis is the most successful marine invasive species in South Africa. Its presence has had significant ecological consequences on the intertidal communities of the west coast. On the south coast, M galloprovincialis co-exists and competes with the indigenous intertidal mussel Perna perna in the lower balanoid zone, where they show partial habitat segregation. The upper and the lower mussel zones are dominated by M. galloprovincialis and P. perna respectively while they co-occur in the mid zone. In this thesis M. galloprovincialis and P. perna are compared in terms of their population genetics and their ecophysiology. The success of an invader depends on its ability to react to new environmental factors, especially when compared to indigenous species. The distribution and diversity of intertidal species throughout the world are strongly influenced by periodic sand inundation and hydrodynamic stress. Occupying the lower intertidal zone, P. perna is more strongly influenced by sand (burial and sand in suspension) than M. galioprovincialis. Despite this, P. perna is more vulnerable to the effects of sand, showing higher mortality rates under experimental conditions in both the laboratory and the field. M. galioprovincialis has longer labial palps than P. perna, indicating a better ability to sort particles. This, and a higher tolerance to anoxia, explains its lower mortality rates when exposed to burial or suspended sand. Habitat segregation is often explained by physiological tolerances, but in this case, such explanations fail. The ability of a mussel to withstand wave-generated hydrodynamic stress depends mainly on its byssal attachment strength. The higher attachment strength of P. perna compared to M. galioprovincialis and of solitary mussels compared to mussels living within a bed (bed mussels) can be explained by more and thicker byssal threads. M galloprovincialis also has a wider shell, is subjected to higher hydrodynamic loads than P. perna and shows a higher theoretical probability of dislodgement, this is borne out under field conditions. The attachment strength of both species increased from higher to lower shore, in parallel to a gradient of a stronger wave action. Monthly measurements showed that P. perna is always more strongly attached than M. galloprovincialis and revealed seasonal fluctuations of attachment strength for both species in response to wave height. The gonad index of both species was negatively cross-correlated with attachment strength. The results are discussed in the context of the evolutionary strategy of the alien mussel, which directs most of its energy to fast growth and high reproductive output, apparently at the cost of reduced attachment strength. This raises the prediction that its invasive impact will be more pronounced at sites subjected to low or moderate wave action at heavily exposed sites. The potential of a species for invasion is also determined by the ability of the invader to disperse. Population genetics provide indirect information about dispersal through a direct measurement of gene flow. The low genetic divergence (measured as mtDNA) of M. galloprovincialis confirms its recent arrival in South Africa. In contrast, the population genetics structure of P. perna revealed strong divergence on the south-east coast, resulting in a western lineage (straddling the distributional gap of the Benguela System), and an eastern lineage, with an overlap region of the two on the south coast between Kenton-on-Sea and Haga Haga. This genetic disjunction may be caused by Agulhas Current acting as an oceanographic barrier to larval dispersal, or by different environmental selective forces acting on regional populations. Over the last ten years, M. galloprovincialis has shown a decrease or cessation of its spread to the east in exactly the region of the genetic disjunction in P. perna, again suggesting either an oceanographic barrier to larval dispersal, or increasing selection driven by sharp gradients in environmental conditions.

No description available.

Organisms inhabit environments that have many dimensions, each of which can vary temporally and spatially. The spatial-temporal variations of environmental stressors and disturbances may have major but different effects on indigenous and invasive species, favouring either of them at different times and places. The invasive mussel Mytilus galloprovincialis invaded the South African coast 30 years ago and, on the south coast of South Africa, it now competes and co-exists with the indigenous Perna perna in the lower eulittoral zone (referred to here as the mussel zone) The invasive and indigenous species dominate the upper and the lower mussel zones respectively, while the two co-exist in the mid-zone. My results show that intertidal mussels experience, and respond to, spatial and temporal fluctuations of several biotic and abiotic stressors. The invasive and the indigenous species adopt different strategies when reacting to environmental factors and their physiological and behavioural responses vary in time and in different habitats as different pressures become of overriding importance. Attachment strength of both species decreased in summer and increased in winter, and was higher on the open coast than in bays for both species, showing a strong positive correlation with wave force in time and space. P. perna had significantly higher attachment strength than M. galloprovincialis but, contrary to previous studies, the difference in gonad index between the two species varied according to the habitat. In bay habitats, M. galloprovincialis had a higher maximum reproductive effort than P. perna, however, on the open coast, there was no significant difference between the two species, suggesting that for the invasive species wave action is a limiting factor not only in terms of the attachment strength but also of energy availability for reproductive tissue development. Major spawning events occurred during periods of low wave action while minor spawning coincided with periods of intense hydrodynamic stress. On the open coast, gonad index was negatively correlated with attachment strength for both species while, in bays, there was no correlation between these two factors for either. The two species also showed different behaviour. In the field, M. galloprovincialis moved significantly more than P. perna over a period of six months. The higher mobility of the invasive species was also confirmed in the laboratory where, in general, M. galloprovincialis formed clumps more readily than P. perna. Taken collectively, these results suggest that channelling more energy into attachment strength limits reproductive tissue development and that, while the indigenous species invests more in byssal production, the invasive species adopts a more dynamic strategy looking for aggregation or a safer arrangement. Higher endolithic infestation and a greater expression of heat shock proteins (Hsps) in mussel populations on the open coast than in bays indicate that this habitat is a more stressful environment not only in terms of wave action. Endolith damaged mussels had significantly lower attachment strengths and condition indices than clean mussels, probably due to the need to channel energy into shell repair. The constant shell repair and expression of Hsps typical of open coast populations are energetically demanding processes. These observations suggest that on the open coast, mussels are subjected to more severe energetic constraints than in bay habitats. Wave and sand stress fluctuated seasonally with the former having a greater effect on mussel mortality on the open coast and the latter a higher impact on bay populations. Overall, mussel mortality rates were higher on the open coast than in bays. My results show that populations on the open coast had fewer private haplotypes and less genetic endemism than those inside bays. Gene flow analysis showed the relatively stable bay habitats act as source populations with greater genetic migration rates out of bays than into them. These differences in genetic structure on scales of las of kilometers show that coastal configuration strongly affects selection, larval dispersal and haplotype diversity. Environmental gradients that are key factors in species distribution over large geographical scales can also be responsible for micro-scale distributions. My results show that M. galloprovincialis colonizes the upper mussel zone where temperature is high, but is less tolerant to this stressor and has to maintain a high expression of Hsps. This suggests that temperature is probably a limiting factor in its invasion towards the sub-tropical east coast. There are inter- and intra-specific differences in responses to the environment which highlight the efforts of M. galloprovincialis and P. perna to optimize resource utilization for survival and reproduction. Determining these differences is crucial to understanding patterns of co-existence between competing indigenous and invasive species.

Dans un contexte de réchauffement climatique et d'anthropisation, les organismes colonisant les espaces côtiers sont, et seront soumis, à des variations importantes de leur milieu de vie. Parmi ces organismes, la moule bleue Mytilus edulis constitue une espèce clé des écosystèmes intertidaux et est utilisée comme sentinelle. Au cours de cette thèse, des moules collectées sur des sites présentant des niveaux de contamination contrastés ont été conditionnées à deux niveaux de température différents avant une exposition à un stress thermique aigu. Des analyses protéomiques ont été réalisées pour explorer les effets de ces stress sur l'homéostasie des protéines. Une forte mortalité est observée exclusivement pour les individus issus du site pollué et conditionnés à des températures modérées. Chez les individus issus du site propre, une abondance de protéines de stress thermique et une sur-expression d'acteurs indiquant la mise en place d'un métabolisme anaérobie est observée. Les individus conditionnés à des températures plus élevées ont une meilleure réponse vis-à-vis des individus conditionnés aux températures modérées. Pour les moules provenant du site contaminé les réponses protéiques sont bien moins nettes et tendent à démontrer un effet additif délétère de la contamination et de la température. Là encore, les individus conditionnés aux températures les plus élevées répondent mieux. En conclusion, les individus au trait de vie non contaminés affichent de meilleures réponses physiologiques que les individus contaminés. De plus, les organismes dont l'historique thermique est favorable, i.e. conditionnés à des températures plus élevées, ont également des réponses améliorées. / Intertidal organisms live in a fluctuating environment. The blue mussel Mytilus edulis is a key species of those ecosystems and are largely use as sentinel species. Global warming associated with anthropization will expose mussels to contaminations together with increased temperatures. In addition, more frequent heatwaves are expected. In this work, mussels were collected at two sites depicting contrasted levels of contamination and thermal exposure in microcosm were conducted. Two acclimation scenarios were set up prior to exposure to an identical acute thermal stress. In order to decipher joint effects of acclimation and contamination on protein homeostasis, gill proteome comparisons were performed. High mortality was observed only for mussels collected at the contaminated site and acclimated to current temperatures. Concerning gill proteome analysis, organisms from the pristine site exhibit high abundance of thermal stress proteins. Proteoforms involved in anaerobic metabolism were also up-regulated. Interestingly, mussels acclimated to the higher temperatures show an enhanced response compare to the one acclimated to current temperatures. Concerning mussels from the contaminated site, the response appears more confusing, excepted for heat stress protein response. This may indicate deleterious effects of combined contamination and heat stress. Therefore, organisms acclimated to higher temperature display improved responses. In conclusion, mussels with a clean life history show better physiological abilities than individuals with contaminated life history. Moreover, organisms prepared to heat stress by higher acclimation temperatures also develop a more effective response.

Mollusks growth is a very important and sensitive response to environmental stresses sincethey are good indicators of the available amount of contaminants in the water; reduced growth represents adverse environmental effects and possible effects on the population.Sweden has about 3,000 Natural Reserves and 28 National Parks. Kosterhavets is the first National Marine Park and it is located on the west coast of Sweden, it is also considered one of the most vulnerable areas since 6000 marine species can be found here and about200 are found nowhere else. It is not only a touristic destination; it is also a home and a work place to many people. The type of pollutants and environmental impacts that areproduced by recreation activities on marinas on such park depend very much on the amount of boats. Mytilus edulis has been widely used to monitor the biological effects of contamination by different ways, such as chemical analysis and biological responses. Themain goal of this project was to assess the status of three different marinas with high and low boat traffic and use the shell length of the blue mussel M. edulis as a potential bioindicator to detect effects from boating activities pressures. The main findings arising from this study are that the sizes of the mussels from the three areas with high boat traffic are significantly smaller than the area with little boat traffic. / <p>www.ima.kth.se</p>

The ability to acclimate to changing temperature has consequences for the biogeographic range of a species and their potential for surviving ocean warming. Using a tide simulator, which recreates tidal conditions by controlling water levels, water and air temperatures, light levels, and food availability, we explored how temperature and tidal zone (i.e. intertidal and subtidal) influences the abundance of sirtuin 5 (SIRT5) protein in a California native mussel (Mytilus californianus). We compared how gill tissue abundance of SIRT5, a key regulator of the cellular stress response and energy metabolism responded in M. californianus exposed to different temperature conditions (13, 16, 19 and 22°C) over a period of four weeks. Two SIRT5 isoforms, one a putative cytosolic form and the other a mitochondrial form were found to be expressed in mussel gill tissue. The mitochondrial isoform increased during acclimation to warm temperatures. This finding is the first to show how SIRT5 protein abundance changes with temperature acclimation. Surprisingly, we did not identify any differences in gill SIRT5 abundance between mussels from intertidal and subtidal locations. Our results suggest that characterizing the responses of SIRT isoforms may lead to a better understanding of the physiological diversity of sirtuins.

La génétique des populations a révélé que la diversité génétique des espèces marines était très souvent distribuée de façon discrète dans l’espace, en mosaïque de patchs populationnels génétiquement homogènes délimités par des discontinuités appelées barrière au flux génique. L’objectif de cette thèse était de contribuer à mieux comprendre les processus expliquant l’origine, le maintien et la position des barrières génétiques au niveau de la zone de transition entre l’Atlantique et la Méditerranée. Dans un premier temps a été étudiée la structure génétique de la moule Mytilus galloprovincialis. Contrairement au cline abrupt et étroit reporté en Espagne, nous avons découvert en Algérie une vaste zone hybride mosaïque sur 600 km de côtes à l'Est du front océanique Almeria-Oran. Dans un deuxième temps a été menée une étude de la structure génétique du gastéropode marin Stramonita haemastoma. Nous avons découvert deux lignées cryptiques différentiellement fixées pour des haplogroupes mitochondriaux, et différenciées sur 3 marqueurs microsatellites développés dans cette thèse. La distribution spatiale en mosaïque est étonnante avec un patch de la lignée atlantique enclavé au nord de la Méditerranée occidentale et bordé par une zone hybride au sud dans la région de Valence. Ces deux études mettent en avant l’importance de l’isolement reproductif intrinsèque dans l’explication de la distribution mosaïque de la diversité génétique marine. Bien que les frontières entre patchs correspondent à des barrières physiques à la dispersion ou à des écotones, l’hydrographie et l’environnement n’expliquent sans doute que la position des discontinuités génétiques mais ni leur origine ni leur maintien. / Population genetics has revealed the genetic diversity of marine species is often subdivided into a mosaic of discrete patches, within which populations are genetically homogeneous, delineated by discontinuities called barriers to gene flow. The aim of this thesis was to contribute to better understand the processes explaining the origin, maintenance and location of genetic barriers at the Atlantic/Mediterranean transition zone. First, we studied the genetic structure of the mussel Mytilus galloprovincialis. In contrast to the abrupt narrow cline reported in Spain, we discovered along the Algerian coastline a 600 km wide mosaic hybrid zone eastward of the Almeria-Oran oceanic front. Second, we studied the genetic structure of a marine gastropod Stramonita haemastoma. We discovered two cryptic lineages differentially fixed for alternative mitochondrial haplogroups, and differentiated at three microsatellite markers developed in this PhD work. Surprisingly, the spatial distribution proved to be an unusual mosaic with a patch of the Atlantic lineage enclaved in the north of the Western Mediterranean Sea, bordered in the South by a hybrid zone in eastern Spain around Valencia. These two studies highlight the importance of intrinsic reproductive isolation in explaining the mosaic distribution of the marine genetic diversity. Although boundaries between patches coincide with physical barriers to dispersal or ecotones, hydrography and environment mainly explain the position of the genetic discontinuities but neither their origin nor their maintenance.