Among efforts to develop sustainable approaches towards the intensive rearing of finfish within open marine waters, is the development of integrated aquaculture techniques. Integrated Multi-Trophic Aquaculture (IMTA), has been promoted as a way to reduce unwanted environmental impacts associated with the intensive production of marine finfish within net-pens. The principle aim of this concept, is the bioremediation of nutrient discharges from fish aquaculture. This is to be achieved by integrating fish cultivation with the growing of species from lower trophic levels, which use the nutrient discharges as a food source. Many studies have been performed that investigate the ability of various species of macroalgae to remove dissolved nutrient discharges, and the ability bivalves to remove solid-bound nutrients, presented as either fish faeces, or an enhanced production of phytoplankton that may be promoted by nutrients emitted by fish-farms. IMTA has also been suggested as a means to improve overall productivity per unit of feed applied to fish, through the conversion of nutrient emissions into additional biomass, such as the tissues of macroalgae or bivalves. Within the research community which focuses upon the environmental impacts of aquaculture, there is a growing awareness that sustainable solutions to aquaculture production cannot be realised through a focus restricted to the growing-phase, and to a limited set of environmental impacts which may this activity may produce. This is because changes to a specific production phase often promote changes at phases located elsewhere along a products value chain. Life-Cycle Assessment (LCA), is a method employed for modelling the environmental impacts that may potentially be generated across the value chain of a product. It is particularly useful for identifying instances of environmental impact shifting; a term used to describe situations where efforts to reduce the contribution of a specific production phase towards one or more environmental impacts, has the effect of either displacing this contribution to another phase, or increases the contribution of production towards different environmental impacts. Despite its apparent suitability, LCA has not previously been fully applied to the environmental impact modelling of open-water IMTA systems. The work presented in the following thesis advances this research front, by using LCA to explore the potential for environmental problem shifting occurring as a consequence of replacing intensive monoculture production, with IMTA. Comprehensive datasets have been acquired from the Chilean aquaculture industry, describing the production of aquafeed and Salmo salar, as well as for the production of the Phaeophytic macroalga, Macrocystis pyrifera, and the molluscan bivalve, Mytilus chilensis. Using LCA methodology, the production of salmon feed, and the production of S.salar, M.pyrifera and M.chilensis, have been assessed for their capacity to contribute towards a variety of global-scale, environmental impacts. IMTA consisting of either S.salar and M.pyrifera, S.salar and M.chilensis, or all three of these species, and combined at ratios required for a bioremediation efficiency of 100 %, 50%, or 20 % of either nitrogen or phosphorous emission from fish, is compared to the monoculture production of S.salar. The comparison is based upon a standardised functional unit, with each species produced through IMTA, being modelled as part of the reference flow required to fulfil the functional unit. Monoculture is compared to IMTA upon the basis of nutritional function, by using a functional unit of mass-adjusted protein content, and mass-adjusted economic value. The use of economic value is based upon the ‘best-case’ assumption, that it serves as a proxy for the total nutritional function that each product offers to human society. The LCAs presented in this study have produced a number of results. Salmon ingredients derived from agricultural crops and animals account for the majority (between 71 % to 98 %) of contributions towards the impacts of compound salmon feed. In general, agricultural crops ingredients contribute more to these impacts than do agricultural animal ingredients, and account for between 31 % and 87 % of the contributions from all ingredients and inputs. In contrast, the combined supply of fish meal and fish oil from capture fisheries is responsible for between 0.13 % and 11 % of all impacts. Vegetable oil accounts for the vast majority of contributions from ingredients derived from agricultural crops. Vegetable oil is modelled as a 50 : 50 blend of sunflower oil and rapeseed, oil, but sunflower oil accounts not only for most of the contributions from vegetable oil, it is responsible for over 50 % of the contributions that all agricultural crop based ingredients contributes towards some impact categories. Replacing sunflower oil with rapeseed oil reduces the contributions of salmon feed by between 6 % and 24 % across 10 out of the 11 impact categories. When compared upon the basis of equal weight, the contributions of fish oil are between 18 % and 99 % lower than those from rapeseed oil. The production of feed is responsible for the majority of contributions (between 32 % and 86 % ) to all impacts of salmon grow-out production. The production of salmon-smolts accounts for between 3 % and 18 %. The majority (64 %) of contributions towards the eutrophication potential of salmon production are from nutrient emissions, which are the result of fish metabolism, whilst nutrients released through the production of feed, the majority of these being from the agricultural production of crop and animals, account for 32 %. Feed production is also a major contributor to the impacts of land-based smolt production, but these contributions (between 12 % and 37 % across all impact categories) are of a lower magnitude than those from the supply of feed to the grow-out phase. Inputs of salt, and inputs of both electricity produced in a diesel power generator and obtained from the national electricity network, are also notable contributors (between 5 % and 67 %, 4 % and 29 %, and 2 % 47 %, respectively) towards the impacts of smolt-production. The main contributors towards the potential impacts of kelp grow-out production (excluding eutrophication potential) are the supply of infrastructure (between 14 % and 89 %), operation of a diesel-powered motorboat for maintenance purposes (between 1 % and 89 %), and the supply ‘of seeded cartridges’ (between 9 and 49 %). The major contributors from the production of ‘seeded cartriges’ in a land-based facility are the supply of electricity from the national electricity network, the supply of fresh water, and the treatment of waste water. The impact potentials of producing seed in this facility might be reduced if the scale of operation is increased. Removal of nitrogen and phosphorous upon the harvesting of kelp is calculated based upon kelp tissue contents of these nutrients. The harvesting of 200 tonnes ha / yr-1, results in a eutrophic potential with a negative value (-376.51 kg of phosphate equivalents). The removal of such a quantity of nutrients might be beneficial if the local marine environment is at risk of hypernutrification, but when no such problem is present, the potential for undesirable consequences of nutrient sequestration should be considered. The major contributor towards the impacts of mussels is the provision of infrastructure (between 25 % and 99.5 %, excluding eutrophication potential). Infrastructure is also responsible for the majority of contributions from mussel seed production. The provision of cotton mesh bags, which are used to aid attatchment of seed to drop-ropes in the grow-out phase, account for between 37% and 99 % of the contributions from the infrastructure from the grow-out phase. This result suggest that either the impacts of mussel production can be reduced by using an alternative material with lower environmental impact potentials, or the inventory data describing the producing of cottonmesh bags requires some improvement. The outcomes of the LCAs of the different IMTA scenarios, are interesting. The results show that choice of species, and the ratios of their combination as required for the different efficiencies of bioremediation, can have a significant effect upon the comparison between IMTA and monoculture. / The study demonstrates a potential for environmental problem shifting as being a consequence of IMTA, especially when the functional unit is mass-adjusted economic value. As bioremediation efficiency increases, contributions towards eutrophication decrease. However, this reduction is achieved at the cost of increasing the contributions of IMTA towards those impact categories, such as ‘ozone layer depletion,’ for which it has a greater contribution than does monoculture. In general, it cannot be concluded from these results that open-water IMTA represents a more sustainable alternative to the monoculture production of Atlantic salmon. The sustainability of IMTA is shown to be dependent upon a variety of trade-offs, between individual environmental impacts, and between these impacts and the nutritional function that the system is capable of providing.

<p>Циљ истраживања је био да се утврди појава и преваленца паразита Marteilia<br />refringens у медитеранској дагњи (Mytilus galloprovincialis) гајеној у<br />Бококоторском заливу. Осим тога имали смо за циљ да утврдимо да ли<br />еколошки фактори (температура морске воде, салинитет, концентрација<br />кисеоника, pH вредност) и присуство бактерија (Escherichia coli и стрептококи<br />фекалног порекла) утичу на појаву овог паразита у медитеранској дагњи.<br />Вршили смо испитивања на шест различитих локација &ndash; узгајалишта<br />медитеранске дагње у Бококоторском заливу. Укупно је узорковано 960 јединки<br />медитеранске дагње. Marteilia refringens је утврђена на четири локације -<br />узгајалишта. Укупна преваленца паразита Marteilia refringens је износила<br />1,25%. Кретала се од 0% на две локације, 0,56% на једној, 1,67% на једној<br />локацији до максимално 3,33% на двe локације. Пратили смо евентуалну<br />појаву паразита током дванаестомесечног периода, а утврдили смо његово<br />присуство од септембра до децембра и у марту месецу. Од праћених<br />еколошких фактора утврдили смо да највећи утицај на појаву паразита има pH<br />вредност морске воде, с тим што се Marteilia refringens појављује при нижој pH<br />вредности од просечне. Утврдили смо и утицај концентрације кисеоника у<br />морској води, при чему се Marteilia refringens јављала при вишим<br />концентрацијама кисеоника. Нисмо утврдили утицај температуре на појаву<br />паразита, али смо забележили појаву паразита при минималној температури<br />од 11,2˚C. Нисмо статистички доказали утицај салинитета и појаве бактерија у<br />морској води на појаву Marteiliae refringens. Доказали смо негативан утицај<br />појаве паразита Marteilia refringens на индекс кондиције медитеранске дагње.<br />Све мартелије утврђене цитолошким и хистолошким испитивањима смо<br />потврдили и молекуларним испитивањима, при чему смо утвдили да наши<br />изолати паразита припадају М типу Marteiliae refringens. Сви н</p> / <p>Cilj istraživanja je bio da se utvrdi pojava i prevalenca parazita Marteilia<br />refringens u mediteranskoj dagnji (Mytilus galloprovincialis) gajenoj u<br />Bokokotorskom zalivu. Osim toga imali smo za cilj da utvrdimo da li<br />ekološki faktori (temperatura morske vode, salinitet, koncentracija<br />kiseonika, pH vrednost) i prisustvo bakterija (Escherichia coli i streptokoki<br />fekalnog porekla) utiču na pojavu ovog parazita u mediteranskoj dagnji.<br />Vršili smo ispitivanja na šest različitih lokacija &ndash; uzgajališta<br />mediteranske dagnje u Bokokotorskom zalivu. Ukupno je uzorkovano 960 jedinki<br />mediteranske dagnje. Marteilia refringens je utvrđena na četiri lokacije -<br />uzgajališta. Ukupna prevalenca parazita Marteilia refringens je iznosila<br />1,25%. Kretala se od 0% na dve lokacije, 0,56% na jednoj, 1,67% na jednoj<br />lokaciji do maksimalno 3,33% na dve lokacije. Pratili smo eventualnu<br />pojavu parazita tokom dvanaestomesečnog perioda, a utvrdili smo njegovo<br />prisustvo od septembra do decembra i u martu mesecu. Od praćenih<br />ekoloških faktora utvrdili smo da najveći uticaj na pojavu parazita ima pH<br />vrednost morske vode, s tim što se Marteilia refringens pojavljuje pri nižoj pH<br />vrednosti od prosečne. Utvrdili smo i uticaj koncentracije kiseonika u<br />morskoj vodi, pri čemu se Marteilia refringens javljala pri višim<br />koncentracijama kiseonika. Nismo utvrdili uticaj temperature na pojavu<br />parazita, ali smo zabeležili pojavu parazita pri minimalnoj temperaturi<br />od 11,2˚C. Nismo statistički dokazali uticaj saliniteta i pojave bakterija u<br />morskoj vodi na pojavu Marteiliae refringens. Dokazali smo negativan uticaj<br />pojave parazita Marteilia refringens na indeks kondicije mediteranske dagnje.<br />Sve martelije utvrđene citološkim i histološkim ispitivanjima smo<br />potvrdili i molekularnim ispitivanjima, pri čemu smo utvdili da naši<br />izolati parazita pripadaju M tipu Marteiliae refringens. Svi n</p> / <p>The aim of the research was to determine the presence and prevalence of the<br />parasite Marteilia refringens in Mediterranean mussels (Mytilus galloprovincialis)<br />cultivated in Boka kotorska Bay. In addition we aimed to determine wheather<br />environmental factors (temperature of seawater, salinity, oxygen concentration, pH<br />value) and the presence of bacteria (Escherichia coli and streptococci of faecal<br />origin) have influence on the occurrence of this parasite in the Mediterranean<br />mussels. We conducted tests at six different locations. In total 960 individuals of<br />Mediterranean mussels were sampled and included in the study. Marteilia refringens<br />was found at four locations. The overall prevalence of the parasite Marteilia<br />refringens was 1.25%. It ranged from 0% at two locations, 0,56% at one, 1.67% at<br />one to 3,33% at two locations. We traced the possible occurrence of the parasite<br />during a twelve month period, and we found its presence from September till<br />December and in March. We found that the pH value of seawater had the greatest<br />impact on the occurence of the parasite from all monitored environmental factors.<br />Marteilia refringens occurs at lower pH values than the average measured pH<br />values of seawater. We also found that Marteilia refringens occured at the higher<br />concentrations of oxygen in the seawater. We did not determine the effect of<br />temperature on the occurrence of the parasite, but we recorded the occurrence of<br />this parasite at a minimum temperature of 11,2˚C. We did not prove that salinity and<br />the presents of bacteria had an influence on the occurrence of Marteilia refringens.<br />We determined a negative impact of Marteilia refringens on the condition index of<br />Mediterranean mussel. All parasites, determined by cytologic and histologic<br />examination were confirmed by molecular methods. We determined that all our<br />isolates belonged to the M type of Marteilia refringens. All our isolates show genetic<br />uniformity, but also show genetic similarity with other isolates of Marteilia</p>

Le suivi de la contamination côtière par les métaux, au moyen de bivalves du genre Mytilus, est de pratique courante dans de nombreux programmes de surveillance à travers le monde. La bioaccumulation résulte d'une interaction entre facteurs physiologiques (croissance, perte de poids, absorption, stockage), chimiques (transferts métaux, spéciation, biodisponibilité) et environnementaux (température, MES, chlorophylle). L'étude et la modélisation de ces interactions est d'un intérêt considérable pour la compréhension des phénomènes et l'interprétation de la bioaccumulation des métaux traces chez la moule. Compte tenu des objectifs de l'étude, afin d'intégrer l'effet de l'état physiologique de l'organisme bioindicateur, le modèle de bioaccumulation choisi suit le concept d'un modèle cinétique à base énergétique, par un couplage d'un modèle de croissance (modèle DEB : à budget énergétique dynamique) à un modèle simple d'accumulation. Afin de le paramétrer et de le calibrer, une étude cinétique d'accumulation (6 mois) et de décontamination (3 mois) a été réalisée pour cinq métaux (Hg, Cd, Pb, Cu et Zn), par la technique des transplants, sur trois sites d'étude aux potentiels trophiques et chimiques différents (Baie du Lazaret, Etang de Bages et l'île de Port-Cros). En plus de traiter de l'accumulation des métaux traces à partir de la voie dissoute et particulaire, le modèle tient compte de la biologie de l'individu et permet de quantifier l'effet de la croissance et des changements environnementaux, tant chimiques que nutritifs, sur la bioaccumulation. L'accumulation des métaux traces se fait à partir de l'eau et/ou de la nourriture alors que l'élimination se fait lors d'étapes physiologiques telles que la reproduction ou directement par excrétion dans le milieu. La contribution de la physiologie de l'organisme détermine des variables d'état tels que le soma, les réserves et le compartiment reproductif, dont les variations dans le temps influencent les cinétiques de capture et d'élimination au sein de l'organisme. Reliant par une méthode explicative les concentrations dans l'organisme à celle du milieu, le modèle permet d'évaluer la contamination effective des sites en s'affranchissant des différences physiologiques dues au trophisme. Cette application a été réalisée sur le réseau de surveillance français RINBIO par analyse inverse.

Mytilus galloprovincialis is invasive on rocky shores on the west coast of South Africa where it has become the dominant intertidal mussel. The success of this species on the west coast and its superior competitive abilities, have led to concern that it may become invasive on the south coast at the expense of the indigenous mussel Perna perna. On shores where these species co-occur, there appears to be habitat segregation among zones occupied by mussels. M.galloprovincialis dominates the high-shore and P.perna the low-shore, with a mixed zone at mid-shore level. This study examined the factors responsible for these differences in distribution and abundance. The study was conducted in Plettenberg Bay and Tsitsikamma (70km apart) on the south coast of South Africa. Each site included two randomly selected locations (300-400m apart). A third mussel species, Choromytilus meridionalis, is found in large numbers at the sand/rock interface at one location in Plettenberg Bay. Aspects of settlement, recruitment, growth and mortality of juvenile and adult mussels were examined at different tidal heights at each site. Quantitative analysis of mussel population structure at these sites supported the initial observation of vertical habitat segregation. Post-larvae were identified to species and this was confirmed using hinge morphology and mitochondrial DNA analysis. Size at settlement was determined for each species to differentiate between primary and secondary settlement. Adult distribution of C.meridionalis was primarily determined by settlement, which was highly selective in this species. Settlement, recruitment and growth of P.perna decreased with increasing tidal height, while post-settlement mortality and adult mortality increased higher upshore. Thus all aspects of P.perna’s life history contribute to the adult distribution of this species. Presumably, the abundance of P.perna on the high-shore is initially limited by recruitment while those that survive remain prone to elimination throughout adulthood. M.galloprovincialis displayed the same patterns of settlement and recruitment as P.perna. However, post-settlement mortality in this species was consistently low in the low and high zones. Juvenile growth also decreased upshore, suggesting that M.galloprovincialis may be able to maintain high densities on the high-shore through the persistence of successive settlements of slow-growing individuals. The low cover of M.galloprovincialis on the lowshore appeared to be determined by adult interactions. M.galloprovincialis experienced significantly higher adult mortality rates than P.perna in this zone. There were seasonal variations in the competitive advantages enjoyed by each species through growth, recruitment or mortality on the low-shore. In summer, P.perna had higher recruitment rates, faster growth and lower mortality rates, while M.galloprovincialis had slightly higher recruitment rates and faster growth rates in winter. P.perna is a warm water species while M.galloprovincialis thrives on the cold-temperate west coast of South Africa. Therefore both species appear to be at the edge of their optimal temperature regimes on the south coast, which may explain the seasonal advantages of each. Nevertheless, P.perna has maintained spatial dominance on the low-shore suggesting that it may ultimately be the winner in competition between these species. M.galloprovincialis appears to have a refuge from competition with P.perna on the high-shore due to its greater tolerance of desiccation stress, while being competitively excluded from the low-shore. Warm water temperatures coupled with poor recruitment rates at most sites may limit the success of M.galloprovincialis on this coast.

Mussels are important ecological engineers on intertidal rocks where they create habitat that contributes substantially to overall biodiversity. They provide secondary substratum for other free-living, infaunal or epifaunal organisms, and increase the surface area for settlement by densely packing together into complex multilayered beds. The introduction of the alien invasive mussel Mytilus galloprovincialis has extended the upper limit of mussels on the south coast of South Africa, potentially increasing habitat for associated fauna. The aim of this study was to describe the structure of mussel beds, the general biodiversity associated with multi- and monolayered mussel beds of indigenous Perna perna and alien M. galloprovincialis, and to determine the relationship between mussels and epibiotic barnacles. This was done to determine the community structure of associated macrofauna and the role of mussels as biological facilitators. Samples were collected in Plettenberg Bay, South Africa, where M. galloprovincialis dominates the high mussel zone and P. perna the low zone. Three 15 X 15 cm quadrats were scraped off the rock in the high and low zones, and in the mid zone where the two mussel species co-exist. The samples were collected on 3 occasions. In the laboratory mussel-size was measured and sediment trapped within the samples was separated through 75 μm, 1 mm and 5 mm mesh. The macrofauna was sorted from the 1 mm and 5 mm sieves and identified to species level where possible. The epibiotic relationship between mussels and barnacles was assessed by measuring the prevalence and intensity of barnacle infestation and the condition index of infested mussels. Multivariate analysis was used on the mean abundance data of the species for each treatment (Hierarchical clustering, multidimensional scaling, analysis of similarity and similarity of percentages) and ANOVA was used for most of the statistical analyses. Overall, the results showed that tidal height influences the species composition and abundance of associated fauna. While mussel bed layering influenced the accumulation of sediments; it had no significant effect on the associated fauna. Time of collection also had a strong effect. While there was an overlap of species among samples from January, May and March, the principal species contributing to similarity among the March samples were not found in the other two months. The outcomes of this study showed that low shore mussel beds not only supported a higher abundance and diversity of species, but were also the most structurally complex. Although the condition index of mussels did not correlate to the percentage cover of barnacle epibionts, it was also evident that low shore mussels had the highest prevalence. The levels of barnacle infestation (intensity) for each mussel species were highest where it was common and lowest where it was least abundant. This is viewed as a natural artefact of the distribution patterns of P. perna and M. galloprovincialis across the shore. Mussels are more efficient as facilitators on the low mussel zone than the high mussel zone possibly because they provide habitats that are more effective in protecting the associated macrofauna from the effects of competition and predation, than they are at eliminating the effects of physical stress on the high shore. Although mussels create less stressful habitats and protect organisms from the physical stress of the high shore, there are clear limitations in their ability to provide ideal habitats. The biological associations in an ecosystem can be made weak or strong depending on the external abiotic factors and the adaptability of the affected organisms.

Recruitment of planktonic larvae into sedentary benthic populations regulates the population dynamics of marine invertebrates. The processes controlling recruitment, however, are poorly understood, and recruitment remains largely unpredictable, which complicates management of exploited shellfish resources. The mussels Perna perna, Choromytilus meridionalis and Mytilus galloprovincialis, found on the south coast of southern Africa, have planktonic larvae and sedentary adult stages. This thesis examines dispersal, settlement and early post-settlement growth and mortality, and their effect on recruitment and demography of intertidal mussel populations in the region of Algoa Bay on the south coast of southern Africa. Temporal and spatial variation in the body mass, density and size structure of mussels, the distribution of bivalve larvae on plankton grids in the nearshore zone and the distribution of a recently introduced invasive mussel, Mytilus galloprovincialis, were examined between 1989 and 1992. Furthermore, data on hourly or daily changes in wind strength and direction, air and sea surface temperatures and low and high tide levels in the study region, were obtained. There were 3-4 peaks in spawning (characterised by an abrupt decline in weight) and settlement activity annually. These peaks varied in exact timing, intensity and duration between sites and over time. However, at a site, spawning was followed by settlement 4-8 weeks later, and there was a significant (P < 0.05) direct correlation between spawning intensity prior to the appearance of a new cohort and the cohort density (settlement intensity). The stochastic spatial and temporal variation in breeding activity was superimposed on a more general pattern of a higher intensity of spawning and settlement in Algoa Bay than on the open coast, and a higher settlement intensity on coastal sandstone shores than on dune rock shores. Spawning was more frequent in winter and spring, and the probability of spawning and settlement peaked around the spring and autumn equinox, if temperature and wind conditions were suitable. Larval behaviour had little effect on their dispersal in the well-mixed nearshore region. Larvae were passively dispersed by currents, and their dispersal range and direction depended on prevailing winds and local topography. The sharp decline in density of recruit and adult M. galloprovincialis with increasing distance from the point of introduction, showed that some larvae were carried by wind generated currents over moderately long distances (-100 km). However, since most (76 %) M. galloprovincialis recruited within 4 km of the parent population, it is possible that larvae become trapped in small gullies and crevices around rocky shores, and have a limited dispersal range. This could explain the link between local patterns of spawning and settlement. The distribution and abundance of settlers on the shore was influenced by larval behaviour and the availability of settlement, substrata. Larvae preferred to settle primarily on foliose coralline algae and migrate to the adult mussel bed when they were larger (0.60-7 mm), but larvae also settled directly on adult mussels, possibly because the amount of coralline algae was limited. Both direct and secondary settlement were considered to be important in maintaining mussel populations since the rate of settlement was low(generally < 60 000.m-2). Cohort analyses showed that prior to maturity post-settlement growth (- 30 mm in 10 months) and mortality rates (60-100%) were high, but varied. When settlement intensity was low this variability uncoupled the relationship between spawning and recruitment intensity. Multiple regression analysis showed that together reproductive effort (gamete output), settlement intensity, growth and mortality prior to maturity, accounted for 76 % of the variance in recruitment into mature adult populations. The low settlement rate coupled with the short life span of mussels « 3 years), meant that populations underwent marked spatial and temporal variations in structure and abundance as settlement intensity varied, but there were consistent general differences between mussel populations on dune rock and sandstone shores in Algoa Bay and on the open coast. It was concluded from these results that, spawning intensity and post-settlement growth and mortality, rather than dispersal, regulated recruitment and the structure and abundance of intertidal P. perna and C. meridionalis populations along the south coast of southern Africa. On the basis of these results it is recommended that species with limited dispersal, variable recruitment and high natural mortality, such as P. perna, should be conserved by protecting a small part of the population in reserves, and controlling utilisation outside reserves to minimize disturbance to local brood stocks. Furthermore, since the potential for reseeding adjacent exploited areas is limited, several small reserves placed at regular intervals along the coast would be more effective than a single large reserve.

Epibiosis is an ecological relationship that has been described as one of the closest possible associations in marine ecosystems. In the space limited rocky intertidal, mussel beds provide important secondary space for barnacles. The epibiotic relationship between mussels and barnacles on the south-east coast of South Africa was considered at different scales, from large-scale, natural patterns of epibiosis on the rocky shore, to fine-scale settlement choices of barnacles and the effects on the condition and growth rates of individual mussels. Mussel and barnacle assemblages were generally stable over a 12-month period. The tracking of individual mussels with and without barnacle epibionts resulted in a significant increase in mortality rate of mussels with epibionts over 12 months (two-way ANOVA, p = 0.028). Barnacles on rocks, as well as on mussels, were also tracked with no significant effect of substratum on mortality of barnacles (two-way ANOVA, p = 0.119). Prevalence and intensity of barnacle infestations was also examined in relation to coastline topography on two co-occurring mussel species, the indigenous Perna perna and invasive Mytilus galloprovincialis. The results were complex, but bay status had significant effects on prevalence and intensity for both mussel species, depending on the time and zone. The effect of bay in relation to time was particularly relevant for M. galloprovincialis (four-way nested ANOVA, Season X Site(Bay): p = 0.0002), where summer prevalence was higher than that of winter in bays, regardless of zone, while in open coast sites, the effect of season was only significant in the mid zone. Patterns of intensity generally showed higher values in summer. Substratum preference by barnacles was investigated by recording settlement, survival and mortality of Chthamalus dentatus barnacles on various treatments. There was a strong preference for the rock-like plastic substratum by primary settlers (pair-wise tests of PERMANOVA: Dead < Rock mimic (p = 0.0001); Replica < Rock mimic (p = 0.019) and Live < Rock mimic (p = 0.0001)). This indicates that barnacles settle on mussel shells only as a secondary choice and that micro-topography is an important variable in barnacle settlement. The effect of barnacle epibiosis on condition index and growth of P. perna and M. galloprovincialis was also examined as a direct indication of the health of mussels subjected to the biological stress of epibiosis. Although not significant (PERMANOVA: P. perna: p(perm) = 0.890; M. galloprovincialis: p(perm) = 0.395), growth for both mussel species was slower for barnacle-infested individuals in summer, which is the main growing season for mussels in the region. Results from condition index calculations, however, showed no negative impacts of epibiotic barnacles (three-way ANCOVA: P. perna: p = 0.372; M. galloprovincialis: p = 0.762). Barnacle epibionts create a new interface between the mussel and its environment and this interaction can affect other members of the community. The possibility of the barnacle epibiont causing increased drag also needs further investigation. Biological processes operating within a wide range of physical stressors drive the interactions on the rocky shore, such as epibiosis. Overall, the results of this study suggest that the epibiotic relationship between mussels and barnacles on the south-east coast of South Africa does not significantly affect the mussel species present and that barnacles only use mussel shells as a secondary choice of substratum.

L’accroissement de la population humaine mondiale a des répercussions majeures, ce n’est donc pas surprenant, compte tenu de la nécessité de nourrir une population grandissante au niveau planétaire, que la production en conchyliculture ait augmenté au cours des dernières décennies. Or, les connaissances acquises concernant les divers facteurs du déterminisme du sexe et du rapport des sexes chez les bivalves sont très limitées et cela pourrait ralentir grandement le taux de production des éleveurs et leur capacité à intervenir si les stocks venaient à diminuer de façon inquiétante. Certains travaux mentionnent que certains facteurs environnementaux, comme la température, auraient un effet sur le rapport des sexes chez une variété de bivalves, incluant la moule bleue commerciale Mytilus edulis, quoiqu’aucune étude n’ait validé cette dernière possibilité. Cela dit, il est possible que l’environnement des adultes puisse aussi affecter le phénotype de la progéniture. En effet, une transmission intergénérationnelle a déjà été identifiée chez Mytilus, mais la possibilité que les conditions des parents affectent le rapport des sexes spécifiquement n’a jamais été abordée. Il est toutefois connu qu’un facteur maternel présent dans l’œuf affecte le sexe de la progéniture et que cette espèce de bivalve a un mode de transmission des mitochondries particulier. Ce mode de transmission appelé « transmission doublement uniparentale » a rendu possible l’identification du sexe chez les embryons. De cette façon, 1938 embryons provenant de 25 croisements artificiels réalisés à trois températures et effectués lors de trois différentes années ont été analysés. Nos analyses mettent en évidence une variation significative dans la proportion de larves femelles entre les années passant de 64 % à 98 %. Dans certains cas, la proportion de femelle varie de 0 à 100 % entre les différents traitements. Même si un effet général sur le rapport des sexes n’était pas significatif, chaque croisement s’est avéré avoir une norme de réaction qui lui est propre face aux 3 différentes températures. Cette étude met en valeur l’effet important de l’environnement sur le déterminisme du sexe chez M. edulis, autant chez les parents que lors du développement des embryons. / The factors affecting sex determination still remain unknown for most bivalve species. Some studies reported that environmental factors, such as temperature, influence sex determination in certain species, and this has been hypothesized also for the blue mussel Mytilus edulis, but not experimentally validated yet. Adult exposure to different environmental conditions during gametogenesis, which occurs seasonally, may also affect offspring phenotype, including sex determination. Intergenerational carryover effects have been reported in bivalves, but the impact of parental exposures on offspring sex determination has not been examined so far. To address these questions, artificial fertilizations were performed on individuals collected in three different years and their embryos and larvae were reared at three different temperatures to specifically test if the environment influence offspring sex ratio through effects on parental developing gametes and/or on developing embryos. We took advantage of the doubly uniparental inheritance of mitochondria in bivalves to determine the sex of the larvae. The analysis of 1938 larvae from 25 crosses revealed that the overall proportion of female larvae was significantly different among years, varying from 64 % to 98 %. While the proportion of female larvae across temperature ranged from 0 to 100 % in some cases, the reaction norms were cross-specific and there were no significant effects of rearing temperature on sex ratio. Taken together, our results suggested that sex determination in M. edulis occur during the gametogenesis according to the genotype of the parents, but could also be changed during the development. More importantly, both processes are strongly affected by environmental conditions.

Les mitochondries sont généralement transmises de façon strictement maternelle. Chez les animaux, il existe une seule exception à ce mode de transmission mitochondriale : la transmission doublement uniparentale (DUI). La DUI est retrouvée uniquement chez certaines espèces de bivalves. Les mâles possèdent dans leurs gamètes le génome mitochondrial paternel, alors que les femelles ont dans leurs oeufs le génome mitochondrial maternel. Ces génomes possèdent respectivement m-orf ou f-orf, un cadre de lecture supplémentaire (outre les 13 codant pour les protéines mitochondriales de référence) potentiellement codant. La présence de ces ORF étant liée au sexe de l’animal, l’hypothèse a été avancée que ces protéines pourraient jouer un rôle dans le maintien de la DUI ou dans le déterminisme sexuel chez ces espèces. Ce projet consiste donc à mieux cerner les fonctions potentielles de ces orfs chez la moule bleue Mytilus edulis. Pour caractériser leur expression, nous avons procédé à des tests d’immunobuvardage sur des lysats de tissus gamétiques et somatiques mâles et femelles, ainsi qu’à des tests d’immunofluorescence sur des cultures cellulaires des deux sexes. Aussi, nous avons effectué des co-immunoprécipitation et des essais pull-down pour préciser les fonctions des protéines par l’entremise des partenaires d’interaction. Nous avons pu observer la présence de M-ORF dans les gonades mâles uniquement, plus particulièrement dans les mitochondries des spermatozoïdes et dans l’acrosome, et ce, uniquement durant la saison de reproduction des moules. F-ORF, cependant, était produite dans tous les tissus à tous les moments de l’année, encore une fois dans les mitochondries des cellules. Les deux protéines ont de nombreux partenaires d’interactions possibles, dont plusieurs sont liés à des processus spécifiques au sexe ou encore aux acides nucléiques. Les protéines M-ORF et F-ORF sont donc bien fonctionnelles. Leurs partenaires potentiels sont multiples, et d’autres essais doivent être effectués afin de préciser les fonctions des protéines. La présence dans l’acrosome de M-ORF est toutefois d’un grand intérêt en lien avec son rôle potentiel dans le DUI et le déterminisme sexuel. / Mitochondria are usually transmitted by strict maternal inheritance. In animals, there is only one exception to this: doubly uniparental inheritance (DUI). DUI can be found only in some bivalve species. Males have in their sperm a paternal mitochondrial genome whereas females have in their eggs the maternal mitochondrial genome. Both genomes possess an orf (other than the 13 coding for annotated mitochondrial proteins) that can potentially code for a protein, called respectively m-orf and f-orf. These genes are sex-specific in gametes, which brought the possibility that there is a link between the orfs and the maintenance of DUI or with sex determination in DUI species. Therefore, this project aims to have a better understanding of the potential functions of these proteins in the blue mussel Mytilus edulis. To demonstrate the proteins' existence, we did Western blot assays on gametic and somatic tissues from males and females, along with immunohistochemistry on cellular cultures of both sexes. To look for possible interaction partners, we did co-immunoprecipitation assays and pull-downs assays. Our results show expression of M-ORF in the male mantle only, more specifically in sperm mitochondria and acrosome. This is found only during the reproductive season of Mytilus edulis. However, F-ORF is expressed in all tissues all year in both sexes, in cells mitochondria. Both proteins have numerous possible interaction partners. Several are linked to sex-specific processes or to interactions with nucleic acids. Both M-ORF and F-ORF are expressed. Potential partners are multiple, and other assays have to be done to further ascertain these proteins' functions. However, the presence of M-ORF in acrosome is of great interest toward a potential function in DUI or in sex determination.

L’épigénétique se rapporte à un ensemble de mécanismes qui modifient l'expression des gènes sans modifier la séquence nucléotidique sous-jacente, produisant une large gamme de variations phénotypiques et répondant aux fluctuations de l'environnement externe ou interne. La méthylation de l'ADN est le processus épigénétique le plus étudié dans les génomes nucléaires des mammifères. Il existe toutefois des lacunes significatives dans la littérature concernant la méthylation de l'ADN mitochondrial (ADNmt), surtout chez les invertébrés. Les bivalves constituent un modèle particulièrement intéressant pour étudier l’épigénétique mitochondriale ou « mitoépigénétique » puisqu’ils possèdent un système de transmission uniparentale double (DUI) de leur mitochondrie, par lequel les mâles héritent des ADNmt paternel (ou mâle ; M) et maternel (ou femelle ; F). La présente étude avait pour objectif de confirmer l’existence de méthylation dans l’ADNmt de la moule bleue (Mytilus edulis), une espèce à DUI. Notre étude a permis de localiser, par immunofluorescence, des cytosines (5mC) et des adénines (6mA) méthylées, ainsi que des méthyltransférases spécifiques aux adénines et aux cytosines, au sein des mitochondries de cette espèce. Ces résultats sont appuyés par une détection de 5mC et de 6mA dans l’ADNmt par digestions enzymatiques, et confirment la présence de méthylation de l'ADNmt chez M. edulis. Nos résultats en immunofluorescence ont également dévoilé un lien entre la présence de 5mC et le stade de développement des gamètes mâles, c’est-à-dire que les gamètes immatures (spermatides) étaient tous méthylés (au niveau mitochondrial) alors qu’une faible proportion de gamètes matures (spermatozoïdes) présentait ce statut de méthylation. Ceci vient corroborer nos résultats enzymatiques, lesquels ont démontré une plus grande variabilité de méthylation entre les mâles qu’entre les femelles. Enfin, nous avons détecté, par séquençage du méthylome de l’ADNmt, un pic de 5mC en contexte non-CpG conservé entre les ADNmt M et F chez les mâles au sein de la région de contrôle de la réplication, soutenant les résultats d’études antérieures chez les vertébrés. Notre étude est la première à démontrer la présence de méthylation dans l’ADNmt d’une espèce DUI, et met en lumière le rôle potentiel de la méthylation de l'ADN dans leur système de transmission mitochondriale. / Epigenetics refers to a set of mechanisms that modify gene expression without altering the underlying nucleotide sequence, producing a wide range of phenotypic variations, and responding to the external or internal environmental fluctuations. DNA methylation is the most extensively studied epigenetic process in mammalian nuclear genomes. However, there are significant gaps in the literature concerning mitochondrial DNA (mtDNA) methylation, especially in invertebrates. Bivalves are a particularly interesting model for studying mitochondrial epigenetics or “mitoepigenetics”, as they possess a doubly uniparental inheritance (DUI) system of their mitochondria, whereby males inherit both paternal and maternal mtDNAs. The aim of this study was to confirm the existence of methylation in the mtDNA of the blue mussel (Mytilus edulis), a DUI species. Using immunofluorescence, we localized methylated cytosines (5mC) and adenines (6mA), as well as adenine- and cytosine-specific methyltransferases, in mitochondria of this species. These results are supported by the detection of 5mC and 6mA in mtDNA by enzymatic digestions, and confirm the presence of mtDNA methylation in M. edulis. Our immunofluorescence results also revealed a link between the presence of 5mC and the developmental stage of male gametes, i.e. immature gametes (spermatids) were all methylated in their mitochondria while only a small proportion of mature gametes (spermatozoa) showed this methylation status. This corroborates our enzymatic results, which demonstrated greater variability in cytosine methylation status among males than among females. Finally, we detected, by mtDNA methylome sequencing, a 5mC peak in non-CpG context conserved between the paternal and maternal mtDNA in males in the region responsible for control of replication (control region), supporting the results obtained in previous studies on vertebrate. Our study is the first to demonstrate the presence of mtDNA methylation in a DUI species, and highlights the potential role of DNA methylation in their mitochondrial transmission system.