Toxicity of environmental and polystyrene plastic particles on the bivalve Corbicula fluminea: focus on the molecular responses.

Among aquatic organisms, filter feeders are particularly exposed to the ingestion of microplastics (MPs) and nanoplastics (NPs). The present study investigates the effect of environmental microplastics (ENV MPs) and nanoplastics (ENV NPs) generated from macro-sized plastic debris collected in the Garonne River (France), and polystyrene NPs (PS NPs) on the freshwater bivalve Corbicula fluminea. Organisms were exposed to plastic particles at three concentrations: 0.008, 10, and 100 µg L-1 for 21 days. Gene expression measurements were conducted in gills and visceral mass at 7 and 21 days to assess the effects of plastic particles on different functions. Our results revealed: (i) an up-regulation of genes, mainly involved in endocytosis, oxidative stress, immunity, apoptosis, and neurotoxicity, at 7 days of exposure for almost all environmental plastic particles and at 21 days of exposure for PS NPs in the gills, (ii) PS NPs at the three concentrations tested and ENV MPs at 0.008 µg L-1 induced strong down-regulation of genes involved in detoxication, oxidative stress, immunity, apoptosis, and neurotoxicity at 7 days of exposure in the visceral mass whereas ENV MPs at 10 and 100 µg L-1 and all ENV NPs induced less pronounced effects, (iii) overall, PS NPs and ENV MPs 0.008 µg L-1 did not trigger the same effects as ENV MPs 10 and 100 µg L-1 and all ENV NPs, either in the gills or the visceral mass at 7 and 21 days of exposure. This study highlighted the need to use MPs and NPs sampled in the environment for future studies as their properties induce different effects at the molecular level to living organisms.

Gamma irradiation-induced offspring masculinization is associated with epigenetic changes in female zebrafish.

Sex ratio variation is a key topic in ecology, because of its direct effects on population dynamics and thus, on animal conservation strategies. Among factors affecting sex ratio, types of sex determination systems have a central role, since some species could have a sex determined by genetic factors, environmental factors or a mix of those two. Yet, most studies on the factors affecting sex determination have focused on temperature or endocrine-disrupting chemicals (EDCs), and much less is known regarding other factors. Exposure to gamma irradiation was found to trigger offspring masculinization in zebrafish. Here we aimed at deciphering the potential mechanisms involved, by focusing on stress (i.e. cortisol) and epigenetic regulation of key genes involved in sex differentiation in fish. Cortisol levels in exposed and control (F0) zebrafish females' gonads were similar. However, irradiation increased the DNA methylation level of foxl2a and cyp19a1a in females of the F0 and F1 generation, respectively, while no effects were detected in testis. Overall, our results suggest that parental exposure could alter offspring sex ratio, at least in part by inducing methylation changes in ovaries.

Trematode genetic patterns at host individual and population scales provide insights about infection mechanisms.

Multiple parasites can infect a single host, creating a dynamic environment where each parasite must compete over host resources. Such interactions can cause greater harm to the host than single infections and can also have negative consequences for the parasites themselves. In their first intermediate hosts, trematodes multiply asexually and can eventually reach up to 20% of the host's biomass. In most species, it is unclear whether this biomass results from a single infection or co-infection by 2 or more infective stages (miracidia), the latter being more likely a priori in areas where prevalence of infection is high. Using as model system the trematode Bucephalus minimus and its first intermediate host cockles, we examined the genetic diversity of the cytochrome c oxidase subunit I region in B. minimus from 3 distinct geographical areas and performed a phylogeographic study of B. minimus populations along the Northeast Atlantic coast. Within localities, the high genetic variability found across trematodes infecting different individual cockles, compared to the absence of variability within the same host, suggests that infections could be generally originating from a single miracidium. On a large spatial scale, we uncovered significant population structure of B. minimus, specifically between the north and south of Bay of Biscay. Although other explanations are possible, we suggest this pattern may be driven by the population structure of the final host.

Sex-specific DNA methylation and transcription of zbtb38 and effects of gene-environment interactions on its natural antisense transcript in zebrafish.

There is increasing evidence for the involvement of epigenetics in sex determination, maintenance, and plasticity, from plants to humans. In our previous work, we reported a transgenerational feminization of a zebrafish population for which the first generation was exposed to cadmium, a metal with endocrine disrupting effects. In this study, starting from the previously performed whole methylome analysis, we focused on the zbtb38 gene and hypothesized that it could be involved in sex differentiation and Cd-induced offspring feminization. We observed sex-specific patterns of both DNA methylation and RNA transcription levels of zbtb38. We also discovered that the non-coding exon 3 of zbtb38 encodes for a natural antisense transcript (NAT). The activity of this NAT was found to be influenced by both genetic and environmental factors. Furthermore, increasing transcription levels of this NAT in parental gametes was highly correlated with offspring sex ratios. Since zbtb38 itself encodes for a transcription factor that binds methylated DNA, our results support a non-negligible role of zbtb38 not only in orchestrating the sex-specific transcriptome (i.e., sex differentiation) but also, via its NAT, offspring sex ratios.

Molecular confirmation of pearl formation in arctic mussels (Mytilus edulis) caused by Gymnophallus bursicola (Odhner 1900) metacercariae.

In recent field studies, suspected gymnophallid metacercariae were histologically located in the mantle of mussels from the Norwegian Sea. Mussels from the sites in which that infection was detected also presented abnormally high pearl numbers. It has been previously described that gymnophallid metacercariae could cause pearl formation processes in mussels, as a host reaction to encapsulate these metacercariae. Given the pathological host reaction these parasites elicit, a study was performed to identify gymnophallid metacercariae found in mussels collected from Tromsø at morphological and molecular level and to assess, by the use of molecular tools, the relationship between the parasite and the biological material inside the pearls. As a result, Gymnophallus bursicola metacercariae infecting Norwegian Mytilus edulis were identified according to morphological characters, along with the first 18S rDNA and COI sequences for this trematode species. In addition, parasite DNA from the core of the pearls was extracted and amplified for the first time, confirming the parasitological origin of these pearls. This procedure could allow identifying different parasitic organisms responsible for the generation of pearls in bivalves.

Altered ovarian transcriptome is linked to early mortality and abnormalities in zebrafish embryos after maternal exposure to gamma irradiation.

Recent laboratory studies focusing on multigenerational approach demonstrated drastic phenotypic effects after chronic fish irradiation exposure. No irradiation effect at phenotypic scale was observed for F0 (reproductive performances) while early mortality and malformations were observed in F1 offspring whether they were irradiated or not. The objective was to study molecular mechanisms likely to be involved in these phenotypic effects induced by parental irradiation. Thus, F0 adult zebrafish were irradiated for ten days until reproduction and maternal involvement in offspring development was assessed. Levels of maternal provided cortisol and vitellogenin, needed for embryo development, were not impacted by irradiation. However, maternal transcriptome highlighted irradiation effect on processes involved in oocyte development, as well as on essential maternal factors needed for offspring development. Therefore, this study highlighted the importance of parental exposure on offspring fate and of the importance of multigenerational exposure in risk assessment.

A New Polyclad Flatworm, Idiostylochus tortuosus gen. nov., sp. nov. (Platyhelminthes, Polycladida) from France. Can this Foreign Flatworm be Responsible for the Deterioration of Oyster and Mussel Farms?

A new species of polyclad flatworm, Idiostylochus tortuosus gen. nov., sp. nov. (Polycladida, Idioplanidae), from Arcachon Bay (France) is described. This description is based on a morphological analysis and a molecular analysis using partial sequences of the 28S and cytochrome Oxidase I (COI) genes. After the molecular analysis Idiostylochus gen. nov. appears to be the second genus of the Family Idioplanidae and closely related to the family Latocestidae as well as the genera Leptostylochus and Mirostylochus. The molecular data revealed that the new species may belong to an Indonesian or Indo-Pacific family, closely related to genera with origins in South Pacific Ocean waters. This species was found feeding on the oysters and mussels of the Arcachon farms.

Two new species of Marphysa (Annelida, Eunicidae) from southern Australia.

Two new species of Marphysa Quatrefages, 1866 are described from the southeast coast of Australia. With the presence of only compound spinigers and the branchiae present over many chaetigers, Marphysa baudini n. sp. belongs to the Sanguinea-group. This species has ventral cirri with an inflated base and digitiform tip and thick and wide anodont pectinate chaetae, with 3-5 internal long and thick teeth. With the presence of only compound falcigers, Marphysa davidattenboroughi n. sp., belongs to the "Aenea-group". This species is characterised by the presence of a bilobed prostomium, a single pair of pygidial cirri and by the presence of thick, and wide anodont pectinate chaetae with 4-6 long internal and thick teeth.

Transgenerational endocrine disruptor effects of cadmium in zebrafish and contribution of standing epigenetic variation to adaptation.

Evidence has emerged that environmentally-induced epigenetic changes can have long-lasting effects on gene transcription across generations. These recent findings highlight the need to investigate the transgenerational impacts of pollutants to assess their long term effects on populations. In this study, we investigated the transgenerational effect of cadmium on zebrafish across 4 generations. A first whole methylome approach carried out on fish of the first two generations led us to focus our investigations on the estradiol receptor alpha gene (esr1). We observed a sex-dependent transgenerational inheritance of Cd-induced DNA methylation changes up to the last generation. These changes were associated with single nucleotide polymorphisms (SNPs) that were themselves at the origin of the creation or deletion of methylation sites. Thus, Cd-induced genetic selection gave rise to DNA methylation changes. We also analyzed the transcription level of various sections of esr1 as well as estrogen responsive genes. While Cd triggered transgenerational disorders, Cd-induced epigenetic changes in esr1 contributed to the rapid transgenerational adaptation of fish to Cd. Our results provide insight into the processes underpinning rapid adaptation and highlight the need to maintain genetic diversity within natural populations to bolster the resilience of species faced with the global environmental changes.

Genetic and epigenetic interplay allows rapid transgenerational adaptation to metal pollution in zebrafish.

Despite still being a matter of debate, there is growing evidence that pollutant-induced epigenetic changes can be propagated across generations. Whereas such modifications could have long-lasting effects on organisms and even on population, environmentally relevant data from long-term exposure combined with follow-up through multiple generations remain scarce for non-mammalian species. We performed a transgenerational experiment comprising four successive generations of zebrafish. Only fish from the first generation were exposed to an environmentally realistic concentration of cadmium (Cd). Using a whole methylome analysis, we first identified the DNA regions that were differentially methylated in response to Cd exposure and common to fish of the first two generations. Among them, we then focused our investigations on the exon 3 (ex3) of the cep19 gene. We indeed recorded transgenerational growth disorders in Cd-exposed fish, and a mutation in this exon is known to cause morbid obesity in mammals. Its methylation level was thus determined in zebrafish from all the four generations by means of a targeted and base resolution method. We observed a transgenerational inheritance of Cd-induced DNA methylation changes up to the fourth generation. However, these changes were closely associated with genetic variations, mainly a single nucleotide polymorphism. This single nucleotide polymorphism was itself at the origin of the creation or deletion of a methylation site and deeply impacted the methylation level of neighboring methylation sites. Cd-induced epigenetic changes were associated with different mRNA transcripts and an improved condition of Cd fish. Our results emphasize a tight relationship between genetic and epigenetic mechanisms and suggest that their interplay and pre-existing diversity can allow rapid adaptation to anthropogenic environmental changes.