The Mediterranean Mussel Mytilus galloprovincialis (Mollusca: Bivalvia) in Chile: Distribution and Genetic Structure of a Recently Introduced Invasive Marine Species.

The genetic characteristics of invasive species have a significant impact on their ability to establish and spread. The blue mussel (Mytilus galloprovincialis), native to the Mediterranean Sea, is a leading invasive species of intertidal coasts throughout much of the world. Here, we used mitochondrial DNA sequence data to investigate the genetic diversity and phylogeographic structure of invasive (M. galloprovincialis) versus native (Mytilus chilensis) populations of blue mussels in Chile. We evaluated whether genetic diversity in invasive populations could be explained by the genetic characteristics of the native sources from which they might be derived. A phylogenetic analysis confirmed two lineages of the invasive M. galloprovincialis, i.e., the NW Atlantic and the Mediterranean lineages. We found no evidence of genetic structure in the invasive range of M. galloprovincialis in Chile, most probably because of its recent arrival. We did, however, detect a spatial mixture of both M. galloprovincialis lineages at sampling locations along the Chilean coast, giving rise to higher levels of genetic diversity in some areas compared to the population of native M. chilensis. The coastal area of the invasion is still small in extent (~100 km on either side of two large ports), which supports the hypothesis of a recent introduction. Further expansion of the distribution range of M. galloprovincialis may be limited to the north by increasing water temperatures and to the south by a natural biogeographic break that may slow or perhaps stop its spread. The use of internal borders as a tool to minimise or prevent M. galloprovincialis spread is therefore a genuine management option in Chile but needs to be implemented rapidly.

Chromosome-Level Genome Assembly of the Blue Mussel Mytilus chilensis Reveals Molecular Signatures Facing the Marine Environment.

The blue mussel Mytilus chilensis is an endemic and key socioeconomic species inhabiting the southern coast of Chile. This bivalve species supports a booming aquaculture industry, which entirely relies on artificially collected seeds from natural beds that are translocated to diverse physical-chemical ocean farming conditions. Furthermore, mussel production is threatened by a broad range of microorganisms, pollution, and environmental stressors that eventually impact its survival and growth. Herein, understanding the genomic basis of the local adaption is pivotal to developing sustainable shellfish aquaculture. We present a high-quality reference genome of M. chilensis, which is the first chromosome-level genome for a Mytilidae member in South America. The assembled genome size was 1.93 Gb, with a contig N50 of 134 Mb. Through Hi-C proximity ligation, 11,868 contigs were clustered, ordered, and assembled into 14 chromosomes in congruence with the karyological evidence. The M. chilensis genome comprises 34,530 genes and 4795 non-coding RNAs. A total of 57% of the genome contains repetitive sequences with predominancy of LTR-retrotransposons and unknown elements. Comparative genome analysis of M. chilensis and M. coruscus was conducted, revealing genic rearrangements distributed into the whole genome. Notably, transposable Steamer-like elements associated with horizontal transmissible cancer were explored in reference genomes, suggesting putative relationships at the chromosome level in Bivalvia. Genome expression analysis was also conducted, showing putative genomic differences between two ecologically different mussel populations. The evidence suggests that local genome adaptation and physiological plasticity can be analyzed to develop sustainable mussel production. The genome of M. chilensis provides pivotal molecular knowledge for the Mytilus complex.

Proximal and fatty acid analysis in Ostrea chilensis, Crassostrea gigas and Mytilus chilensis (Bivalvia: Mollusca) from southern Chile.

Oysters and blue mussels are important hydrobiological resources for aquaculture. In Chile, they are farming on the Chiloé island, where around 18% of the world's mussels are produced, however, their nutritional dynamics are largely unknown. For this reason, the objective of this study was to determine the proximal biochemical composition and the fatty acid profile in the Chilean oyster (Ostrea chilensis), the Pacific oyster (Crassostrea gigas) and the Chilean mussel (Mytilus chilensis), to perform an intra and interspecific comparison. Shellfish sampled in winter were characterized by a high protein content, followed by medium values for lipid content and a low carbohydrate content compared to similar species in Europe. Also, oysters and mussels were found to be rich in omega-3 long chain polyunsaturated fatty acid (n-3 LC-PUFA), so they can be considered excellent functional food option for a healthy human diet. Their high contribution of n-3 LC-PUFA ranged between 5.2-12.9 µg FA mg-1 dry weight with high n-3/n-6 ratios, which depends on both the species and the on-growing location. Both taxa can be considered a plausible option to promote a healthy diet of marine origin in future generations. Also, these results could benefit the projection and development of aquaculture of these mollusks.

Feeding response and dynamic of intoxication and detoxification in two populations of the flat oyster Ostrea chilensis exposed to paralytic shellfish toxins (PST).

Juvenile oysters (Ostrea chilensis) from two populations (Quempillén estuary and Pullinque bay) were exposed to a toxic diet containing paralytic shellfish toxins (PST), produced by Alexandrium catenella, followed by a detoxification period. Feeding behaviour, toxin profile, dynamics of intoxication/detoxification, and survival were evaluated over the entire experimental period. Both populations reduced their feeding rates during the 30-day exposure to the toxic diet. This negative effect was reversible when the diet was switched to the non-toxic one. Oysters from the estuary accumulated PST more rapidly than the population from the bay, suggesting their increased ability to cope with more adverse conditions. Both populations showed low detoxification capacity. Survival was significantly higher in oysters from the estuary, compared to those from the bay. Due to the increasing frequency and intensity of A. catenella blooms in southern Chile, it is necessary to better understand the responses of O. chilensis in different environments. This is important not only because of the ecological and commercial relevance of the bivalve, but also in consideration of expected climate change scenarios, where the new environmental conditions could favour the frequency and intensity of harmful algal bloom events.

Genetic structure and diversity of the Chilean flat oyster Ostrea chilensis (Bivalvia: Ostreidae) along its natural distribution from natural beds subject to different fishing histories.

Ostrea chilensis (Küster, 1844), the flat oyster, is native to Chile and New Zealand. In Chile, it occurs in a few natural beds, from the northern part of Chiloé Island (41 ºS) to the Guaitecas Archipelago (45 ºS). This bivalve is slow growing, broods its young, and has very limited dispersal potential. The Ostrea chilensis fishery has been over-exploited for a number of decades such that in some locations oysters no longer exist. The aim of this study was to study the genetic diversity of the Chilean flat oyster along its natural distribution to quantify the possible impact of the dredge fishery on wild populations. The genetic structure and diversity of Ostrea chilensis from six natural beds with different histories of fishing activity were estimated. Based on mitochondrial (Cytb) and nuclear (ITS1) DNA sequence variation, our results provide evidence that genetic diversity is different among populations with recent history of wild dredge fishery efforts. We discuss the possible causes of these results. Ultimately, such new information may be used to develop and apply new management measures to promote the sustainable use of this valuable marine resource.

Blue mussels of the Mytilus edulis species complex from South America: The application of species delimitation models to DNA sequence variation.

Smooth-shelled blue mussels, Mytilus spp., have a worldwide antitropical distribution and are ecologically and economically important. Mussels of the Mytilus edulis species complex have been the focus of numerous taxonomic and biogeographical studies, in particular in the Northern hemisphere, but the taxonomic classification of mussels from South America remains unclear. The present study analysed 348 mussels from 20 sites in Argentina, Chile, Uruguay and the Falkland Islands on the Atlantic and Pacific coasts of South America. We sequenced two mitochondrial locus, Cytochrome c Oxidase subunit I (625 bp) and 16S rDNA (443 bp), and one nuclear gene, ribosomal 18S rDNA (1770 bp). Mitochondrial and nuclear loci were analysed separately and in combination using maximum likelihood and Bayesian inference methods to identify the combination of the most informative dataset and model. Species delimitation using five different models (GMYC single, bGMYC, PTP, bPTP and BPP) revealed that the Mytilus edulis complex in South America is represented by three species: native M. chilensis, M. edulis, and introduced Northern Hemisphere M. galloprovincialis. However, all models failed to delimit the putative species Mytilus platensis. In contrast, however, broad spatial scale genetic structure in South America using Geneland software to analyse COI sequence variation revealed a group of native mussels (putatively M. platensis) in central Argentina and the Falkland Islands. We discuss the scope of species delimitation methods and the use of nuclear and mitochondrial genetic data to the recognition of species within the Mytilus edulis complex at regional and global scales.

Lethal and sub-lethal effects of commonly used anti-sea lice formulations on non-target crab Metacarcinus edwardsii larvae.

The pesticides used by the salmon industry to treat sea lice, are applied in situ via a bath solution and are subsequently discharged into the surrounding medium. The effects of cypermethrin, deltamethrin, azamethiphos and hydrogen peroxide were assessed on the performance of Metacarcinus edwardsii larvae, an important crab for Chilean fishery. All larvae were dead or dying after 30 min of exposure to cypermethrin and after 40 min to deltamethrin at concentrations 100 and 20 times lower (0.15 and 0.1 µg L-1, respectively) than the concentrations and exposure times recommended by the manufacturers (CRM) to treat sea lice. Azamethiphos affected all larvae at a concentration 10 times lower than CRM. Hydrogen peroxide had the lowest detrimental effects, but at the CRM, 100% of the larvae were affected. Sub-lethal effects, i.e prolonged developmental time, were observed at concentrations lower than CRM. Repeated exposure to azamethiphos (0.0625-0.5 µg L-1) and hydrogen peroxide (188-1500 mg L-1) had effects on survival. In conclusion, the pesticides used against parasitic copepod tested here, negatively affect non-target crustacean larvae. Due to the product's characteristics, the lethal effects of the pyrethroids probably are restricted to the time and area of application, while the action of azamethiphos may extend to a wider area. Current data are insufficient to accurately dimension the effects of these compounds in the field. More research is required to evaluate the consequences of prolonged developmental times and/or reduction in appendage mobility, so as the effects of these compounds on the pelagic and benthic communities.

Molecular detection and species identification of Alexandrium (Dinophyceae) causing harmful algal blooms along the Chilean coastline.

BACKGROUND AND AIMS: On the basis of morphological evidence, the species involved in South American Pacific coast harmful algal blooms (HABs) has been traditionally recognized as Alexandrium catenella (Dinophyceae). However, these observations have not been confirmed using evidence based on genomic sequence variability. Our principal objective was to accurately determine the species of Alexandrium involved in local HABs in order to implement a real-time polymerase chain reaction (PCR) assay for its rapid and easy detection on filter-feeding shellfish, such as mussels. METHODOLOGY: For species-specific determination, the intergenic spacer 1 (ITS1), 5.8S subunit, ITS2 and the hypervariable genomic regions D1-D5 of the large ribosomal subunit of local strains were sequenced and compared with two data sets of other Alexandrium sequences. Species-specific primers were used to amplify signature sequences within the genomic DNA of the studied species by conventional and real-time PCR. PRINCIPAL RESULTS: Phylogenetic analysis determined that the Chilean strain falls into Group I of the tamarensis complex. Our results support the allocation of the Chilean Alexandrium species as a toxic Alexandrium tamarense rather than A. catenella, as currently defined. Once local species were determined to belong to Group I of the tamarensis complex, a highly sensitive and accurate real-time PCR procedure was developed to detect dinoflagellate presence in Mytilus spp. (Bivalvia) samples after being fed (challenged) in vitro with the Chilean Alexandrium strain. The results show that real-time PCR is useful to detect Alexandrium intake in filter-feeding molluscs. CONCLUSIONS: It has been shown that the classification of local Alexandrium using morphological evidence is not very accurate. Molecular methods enabled the HAB dinoflagellate species of the Chilean coast to be assigned as A. tamarense rather than A. catenella. Real-time PCR analysis based on A. tamarense primers allowed the detection of dinoflagellate DNA in Mytilus spp. samples exposed to this alga. Through the specific assignment of dinoflagellate species involved in HABs, more reliable preventive policies can be implemented.

Allozymic variation and differentiation in the chilean blue mussel, Mytilus chilensis, along its natural distribution

Genetic differentiation in the Chilean blue mussel Mytilus chilensis (Hupé 1854) was investigated based on the variation in the allozyme frequencies of Pgm, Gpi, Icd, Me, Gsr, Lap and Pep in eight samples collected along 1800 km from Arauco (VIII Region) to Punta Arenas (XII Region). Despite the large geographic separations, values of Neis unbiased genetic distance, D (0.004-0.048) and standardised genetic variation among populations, Fst (0.011-0.055) were small. The levels of gene flow (Nm = 8) found in this study prevent the effect of differentiation among populations by genetic drift. This findings indicate that its long-lived planktotrophic larvae provides this species with considerable dispersal ability throughout its range which is favoured by the ocean currents along the chilean coast. In terms of management of the M. chilensis fishery, the results provide no evidence for discrete stocks, with the possible exception of the Punta Arenas population. Considering the intensive aquaculture activities with this species the present study provide preliminary data which can be used as a baseline for further characterization and /or monitoring these mussel populations.