Metallothionein gene identification and expression in the cockle ( Cerastoderma edule) under parasitism (trematodes) and cadmium contaminations

Parmi les organismes benthiques, les bivalves sont souvent utilisés comme bioindicateurs des pollutions environnementales, du fait de leur importante capacité de bioaccumulation des métaux lourds conduisant à l'induction des métallothionéines (MT). Les MT sont de petites protéines cytosoliques capables de fixer les métaux et qui sont impliquées dans l'homéostasie et la détoxication de ces métaux chez les organismes. Ces protéines peuvent être induites par un grand nombre de facteurs tels que les hormones, les stress physiques, le parasitisme. La quantification des MT en relation avec le parasitisme est rarement reportée dans la littérature, bien que le parasitisme soit omniprésent et très délétère chez les bivalves. De plus, seulement un petit nombre de gènes de MT ont été identifiés chez les mollusques. Cette étude décrit la caractérisation de la séquence partielle d'un gène de MT (Cemt1) chez la coque Cerastoderma edule, qui présente plus de 80 % d'homologie avec de nombreuses séquences de MT de bivalves. A partir de cette séquence, des amorces spécifiques, utilisables en PCR quantitative en temps réel, ont été déterminées. Le niveau d'expression du gène Cemt1, ainsi que la quantité de protéine MT ont été analysés chez des individus soumis à différents paramètres : sans parasites ou infestés par le trématode digène Himasthla elongata, et lors d'expositions au cadmium à 15 µg Cd L−1. Des résultats concordants ont été obtenus lors de la quantification des protéines MT et de l'analyse de l'expression du gène Cemt1. Ceux-ci mettent en évidence que les concentrations en MT augmentent significativement par les deux types de traitement (infestation parasitaire et exposition au cadmium).Among benthic organisms, bivalves are often used as bioindicators of environmental pollution because of their high bioaccumulation capacities for heavy metals leading to metallothioneins (MT) induction. MT are small cytosolic metal-binding proteins involved in metal homeostasis and detoxification in living organisms. These proteins can also be induced by a wide range of factors, such as hormones, physical stress, parasitism. MT quantification in relation to parasitism is rarely reported in literature, while parasites are omnipresent and have deleterious impacts on bivalves. Moreover, only a few number of MT genes have been characterized in molluscs. This study describes the partial sequence of the MT gene (Cemt1) in the edible cockle Cerastoderma edule. The cockle's MT cDNA was sequenced and showed over 80% homology to several other bivalve MT sequences. This sequence was then used to determine MT specific primers which can be used in quantitative real time PCR. MT protein and gene expression levels were quantified for individuals selected under different conditions: free from or infected by the digenean trematode Himasthla elongata, and under cadmium exposure at 15 mu g Cd L-1. Results evidenced that MT concentrations were significantly increased by both treatments; parasite infection and Cd exposure. Moreover, congruent results between MT protein and gene expression levels were obtained

Cockle infection by Himasthla quissetensis – II. The theoretical effects of climate change

Numerous marine populations experience parasite pressure. This is the case of the cockles Cerastoderma edule which are often highly infected by trematode macroparasites. These parasites display a complex life cycle, with a succession of free-living and parasitic stages. Climate, and in particular temperature, is an important modulator of the transmission dynamics of parasites. Consequently, global change is thought to have implications for the epidemiology of infectious diseases. Using Himasthla quissetensis, a dominant parasite of cockles as 2nd intermediate host in Arcachon Bay (France), we used mathematical models of parasite emergence (cercariae) and parasite infection (metacercariae) in cockles as a function of water temperature, in order to study different scenarios of temperature increases. Globally, with a + 0.5 °C to + 6.0 °C simulation, cumulated emergence of cercariae and accumulation of metacercariae tended to decrease or stagnate, respectively. This is the consequence of a trade-off between sooner (spring) and later (autumn) cercariae emergence/infestation on one hand, and a longer inhibition period of cercariae emergence/infestation during the hottest days in summer. Using sea water temperature in Oualidia (Morocco) where mean annual sea temperature is 3 °C higher than in Arcachon Bay, our model predicted infestation all year long (no seasonality). The model gave a correct estimation of the total number of parasites that was expected in cockles. Conversely, observed infestation in Oualidia followed a seasonal pattern like in Arcachon Bay. These results suggest that, if temperature is a strong driver of parasite transmission, extrapolation in the framework of climate change should be performed with caution