Time-series incubations in a coastal environment illuminates the importance of early colonizers and the complexity of bacterial biofilm dynamics on marine plastics.

The problematic of microplastics pollution in the marine environment is tightly linked to their colonization by a wide diversity of microorganisms, the so-called plastisphere. The composition of the plastisphere relies on a complex combination of multiple factors including the surrounding environment, the time of incubation along with the polymer type, making it difficult to understand how the biofilm evolves during the microplastic lifetime over the oceans. To better define bacterial community assembly processes on plastics, we performed a 5 months spatio-temporal survey of the plastisphere in an oyster farming area in the Bay of Brest (France). We deployed three types of plastic pellets in two positions in the foreshore and in the water column. Plastic-associated biofilm composition in all these conditions was monitored using 16 S rRNA metabarcoding and compared to free-living and attached bacterial members of seawater. We observed that bacterial families associated to plastic pellets were significantly distinct from the ones found in seawater, with a significant prevalence of filamentous Cyanobacteria on plastics. No convergence towards a unique plastisphere was detected between polymers exposed in the intertidal and subtidal area, emphasizing the central role of the surrounding environment on constantly shaping the plastisphere community diversity. However, we could define a bulk of early-colonizers of marine biofilms such as Alteromonas, Pseudoalteromonas or Vibrio. These early-colonizers could reach high abundances in floating microplastics collected in field-sampling studies, suggesting the plastic-associated biofilms could remain at early development stages across large oceanic scales. Our study raises the hypothesis that most members of the plastisphere, including putative pathogens, could result of opportunistic colonization processes and unlikely long-term transport.

Cg-IkappaB, a new member of the IkappaB protein family characterized in the pacific oyster Crassostrea gigas.

Inhibitors of NF-kappaB (IkappaBs) have been implicated as major components of the Rel/NF-kappaB signaling pathway, which is an important mediator of immune responses throughout much of the animal kingdom. Here we report the first characterization of a bivalve mollusc cDNA that encodes a novel IkappaB member called Cg-IkappaB showing the conservation of most IkappaB protein characteristics. Sequences as well as phylogenetic analyses reveal a high level of identity between not only Cg-IkappaB and other mollusc and insect IkappaB-like proteins but also similarities to vertebrate IkappaB-alpha and -epsilon isoforms. Expression analyses demonstrated that the transcript is widely expressed in all oyster tissues. This work is consistent with our previous discovery of several members of an NF-kappaB pathway in Crassostrea gigas and further sustains the hypothesis of a conserved scheme of immune gene regulation through most of the metazoans.

Evidence for the involvement of pathogenic bacteria in summer mortalities of the Pacific oyster Crassostrea gigas.

A study was conducted to investigate the involvement of bacteria in oyster mortalities during summer. Moribund and apparently healthy oysters were sampled during mortality events along the French coast and in rearing facilities, usually when temperature reached 19 degrees C or higher, and oysters were in the gonadal maturation phase. Hemolymph samples were aseptically withdrawn and submitted to bacteriological analysis. In healthy oysters, bacteria colonized hemolymph at low concentrations depending on the location. In most moribund oysters, bacteria were present in hemolymph and other tissues. These bacterial populations were more often diverse in oysters originating from the open sea than from facilities where animals were generally infected by a single type of bacterium. Only the dominant colonies were identified by phenotypic and genotypic characters (RFLP of GyrB gene and partial sequence of 16S rRNA gene). They belonged to a limited number of species including Vibrio aestuarianus, members of the V. splendidus group, V. natriegens, V. parahaemolyticus, and Pseudoalteromonas sp. The most frequently encountered species was V. aestuarianus (56% of isolates), which was composed of several strains closely related by their 16S rRNA gene but diverse by their phenotypic characters. They appeared intimately linked to oysters. The species within the V. splendidus group were less prevalent (25% of isolates) and more taxonomically dispersed. A majority of the dominant strains of V. aestuarianus and V. splendidus group injected to oysters induced mortality, whereas others belonging to the same species, particularly those found in mixture, appeared innocuous.

Response of Penaeus indicus females at two different stages of ovarian development to a lethal infection with Vibrio penaeicida.

An association between vitellogenesis and the immune system was suggested in crustaceans from studies on plasma lipoproteins. The present research studies the effect of an experimentally induced bacterial infection on vitellogenesis in females of the shrimp Penaeus indicus, as a model for penaeid species. Pre-vitellogenic and vitellogenic P. indicus females were experimentally infected with an extremely pathogenic bacterium, Vibrio penaeicida. The peak in mortality occurred earlier in pre-vitellogenic animals than in vitellogenic ones, although the final mortality level ( approximately 64-74%) 52h post-infection was nearly the same for the two groups. Twenty hours after infection, the total number of haemocytes was significantly reduced in vitellogenic females while there was no change in the pre-vitellogenic group. Protein synthesis in ovaries was not significantly affected by infection, at the two stages of ovarian development. No differences were found in mRNA levels of shrimp ovarian peritrophin protein (SOP), but preliminary results showed that mRNA expression of vitellin (VT) was reduced in a heavily infected vitellogenic female. The total amount of lipids in the haemolymph of vitellogenic females was almost twice higher than that of pre-vitellogenic ones. However, there was no change in the total content of lipids, lipid classes and fatty acid distribution in haemolymph or hepatopancreas following infection. Although vitellogenic and pre-vitellogenic females probably respond differently to a lethal bacterial infection, physiological differences may be concealed by the rapid onset of mortality.