A simulation study evaluating how population survival and genetic diversity in a newly established population can be affected by propagule size, extinction rates, and initial heterozygosity.

The introduction and establishment of invasive species in regions outside their native range, is one of the major threats for the conservation of ecosystems, affecting native organisms and the habitat where they live in, causing substantial biological and monetary losses worldwide. Due to the impact of invasive species, it is important to understand what makes some species more invasive than others. Here, by simulating populations using a forward-in-time approach combining ecological and single polymorphic nucleotides (SNPs) we evaluated the relation between propagule size (number of individuals = 2, 10, 100, and 1,000), extinction rate (with values 2%, 5%, 10%, and 20%), and initial heterozygosity (0.1, 0.3, and 0.5) on the population survival and maintenance of the heterozygosity of a simulated invasive crab species over 30 generations assuming a single introduction. Our results revealed that simulated invasive populations with initial propagule sizes of 2-1,000 individuals experiencing a high extinction rate (10-20% per generation) were able to maintain over 50% of their initial heterozygosity during the first generations and that under scenarios with lower extinction rates invasive populations with initial propagule sizes of 10-1,000 individuals can survive up to 30 generations and maintain 60-100% of their initial heterozygosity. Our results can help other researchers better understand, how species with small propagule sizes and low heterozygosities can become successful invaders.

Natural mega disturbances drive spatial and temporal changes in diversity and genetic structure on the toadfish Aphos porosus.

Natural disturbances can modify extinction-colonization dynamics, driving changes in the genetic diversity and structure of marine populations. Along Chilean coast (36°S, 73°W), a strong hypoxic-upwelling event in 2008, and a mega earthquake-tsunami in 2010 caused mass mortality within the Aphos porosus population, which is a vulnerable species with low dispersal potential. We evaluated the effects of these two major disturbances on the diversity and spatial-temporal genetic structure of Aphos porosus in two neighboring areas that were impacted on different levels (High level: Coliumo Bay; Low level: Itata Shelf). Thirteen microsatellites (from 2008 to 2015) amplified in individuals collected from both locations were used to evaluate the effects of the two disturbances. Results showed that after the strong hypoxic-upwelling event and the mega earthquake-tsunami, Aphos porosus populations exhibited lower genetic diversity and less effective population sizes (Ne < 20), as well as asymmetries in migration and spatial-temporal genetic structure. These findings suggest a rise in extinction-recolonization dynamics in local Aphos porosus populations after the disturbances, which led to a loss of local genetic diversity (mainly in Coliumo Bay area impacted the most), and to greater spatial-temporal genetic structure caused by drift and gene flow. Our results suggest that continuous genetic monitoring is needed in order to assess potential risks for Aphos porosus in light of new natural and anthropogenic disturbances.

Intraspecific variation in reproductive traits and embryo elemental composition of the crab Hemigrapsus crenulatus (Milne Edwards, 1837) across fluctuating coastal environments along Chilean coasts.

The estuarine crab Hemigrapsus crenulatus has a wide distribution range along the coast of the South Pacific Ocean (from 20 °S to 53 S°). This decapod is abundant in these coastal and estuarine habitats and plays a key ecological role as prey of the snook fish (Eleginops maclovinus) and kelp gull (Larus dominicanus). Its diet consists of detritus, dead fish, and crustaceans, and the macroalgae Ulva sp. In response to contrasting environmental conditions and anthropogenic impacts along the Chilean coast, H. crenulatus may present intraspecific variations in its reproductive traits and changes in the elemental composition of its embryos, directly affecting its biological fitness. Along the Chilean coast, female individuals were collected during late spring 2019 and early summer 2020 (from November 2019 to February 2020) in six areas of Chile (north: Tongoy (30°S); south-central: Lenga (36°S), Tubul (37°S); south austral: Calbuco (41°S), Castro (42°S), Quellón (43°S). The environmental conditions (i.e. sea surface temperature, precipitation and chlorophyll α) present during each sampling event were also recorded. We evaluated the reproductive parameters of females (fecundity, reproductive output (RO)) as well as their body size (carapace width and dry weight), volume, water content, dry weight, elemental composition (carbon, hydrogen, nitrogen: CHN), and finally the energy content of their embryos. Our results indicated that the environmental conditions of the sea water temperature, precipitation (proxy of water salinity) and chlorophyll α (proxy of food availability) have direct effects on the reproductive parameters of females and the characteristics of their embryos. We observed a low fecundity and high RO in Calbuco and Quellón, where precipitation was high (i.e. diluted salinity) and temperatures and productivity was low. For embryo traits, the highest values of volume and water content were observed in female crabs from the estuarine areas (i.e. Tongoy, Lenga, Tubul), values much higher than those found in the internal sea of Chile (i.e. Calbuco, Castro, Quellón). For the elemental composition, we observed high nitrogen levels and a low C:N proportion in embryos from female crabs from Lenga (a nitrogen enriched area). Fluctuating environmental variables among localities proved to modulate intraspecific variations in females and embryos of H. crenulatus, revealing different reproductive strategies, particularly in the quality and energy investment per embryo, which subsequently influenced successful embryogenesis and larval survival.

Predator control of marine communities increases with temperature across 115 degrees of latitude.

Early naturalists suggested that predation intensity increases toward the tropics, affecting fundamental ecological and evolutionary processes by latitude, but empirical support is still limited. Several studies have measured consumption rates across latitude at large scales, with variable results. Moreover, how predation affects prey community composition at such geographic scales remains unknown. Using standardized experiments that spanned 115° of latitude, at 36 nearshore sites along both coasts of the Americas, we found that marine predators have both higher consumption rates and consistently stronger impacts on biomass and species composition of marine invertebrate communities in warmer tropical waters, likely owing to fish predators. Our results provide robust support for a temperature-dependent gradient in interaction strength and have potential implications for how marine ecosystems will respond to ocean warming.

Greater functional similarity in mobile compared to sessile assemblages colonizing artificial coastal habitats.

Among anthropogenic habitats built in the marine environment, floating and non-floating structures can be colonized by distinct assemblages. However, there is little knowledge whether these differences are also reflected in the functional structure. This study compared the functional diversity of sessile and mobile invertebrate assemblages that settle over three months on floating vs. non-floating artificial habitats, in two Chilean ports. Using morphological, trophic, behavioral, and life history traits, we found differences between mobile and sessile assemblages regarding the effect of the type of habitat on the functional diversity. Compared to sessile assemblages, a greater functional similarity was observed for mobile assemblages, which suggests that their dispersal capacity enables them to balance the reduced connectivity between settlement structures. No traits, prevailing or selected in one or the other habitat type, was however clearly identified; a result warranting for further studies focusing on more advanced stages of community development.

RNA-Seq reveals divergent gene expression between larvae with contrasting trophic modes in the poecilogonous polychaete Boccardia wellingtonensis.

The polychaete Boccardia wellingtonensis is a poecilogonous species that produces different larval types. Females may lay Type I capsules, in which only planktotrophic larvae are present, or Type III capsules that contain planktotrophic and adelphophagic larvae as well as nurse eggs. While planktotrophic larvae do not feed during encapsulation, adelphophagic larvae develop by feeding on nurse eggs and on other larvae inside the capsules and hatch at the juvenile stage. Previous works have not found differences in the morphology between the two larval types; thus, the factors explaining contrasting feeding abilities in larvae of this species are still unknown. In this paper, we use a transcriptomic approach to study the cellular and genetic mechanisms underlying the different larval trophic modes of B. wellingtonensis. By using approximately 624 million high-quality reads, we assemble the de novo transcriptome with 133,314 contigs, coding 32,390 putative proteins. We identify 5221 genes that are up-regulated in larval stages compared to their expression in adult individuals. The genetic expression profile differed between larval trophic modes, with genes involved in lipid metabolism and chaetogenesis over expressed in planktotrophic larvae. In contrast, up-regulated genes in adelphophagic larvae were associated with DNA replication and mRNA synthesis.

Interpopulational differences in the nutritional condition of Aequiyoldia eightsii (Protobranchia: Nuculanidae) from the Western Antarctic Peninsula during austral summer.

The Western Antarctic Peninsula (WAP) is a hotspot for environmental change and has a strong environmental gradient from North to South. Here, for the first time we used adult individuals of the bivalve Aequiyoldia eightsii to evaluate large-scale spatial variation in the biochemical composition (measured as lipid, protein and fatty acids) and energy content, as a proxy for nutritional condition, of three populations along the WAP: O'Higgins Research Station in the north (63.3°S), Yelcho Research Station in mid-WAP (64.9°S) and Rothera Research Station further south (67.6°S). The results reveal significantly higher quantities of lipids (L), proteins (P), energy (E) and total fatty acids (FA) in the northern population (O'Higgins) (L: 8.33 ± 1.32%; P: 22.34 ± 3.16%; E: 171.53 ± 17.70 Joules; FA: 16.33 ± 0.98 mg g) than in the mid-WAP population (Yelcho) (L: 6.23 ± 0.84%; P: 18.63 ± 1.17%; E: 136.67 ± 7.08 Joules; FA: 10.93 ± 0.63 mg g) and southern population (Rothera) (L: 4.60 ± 0.51%; P: 13.11 ± 0.98%; E: 98.37 ± 5.67 Joules; FA: 7.58 ± 0.48 mg g). We hypothesize these differences in the nutritional condition could be related to a number of biological and environmental characteristics. Our results can be interpreted as a consequence of differences in phenology at each location; differences in somatic and gametogenic growth rhythms. Contrasting environmental conditions throughout the WAP such as seawater temperature, quantity and quality of food from both planktonic and sediment sources, likely have an effect on the metabolism and nutritional intake of this species.

Rapid recovery of native habitat-builders following physical disturbance on pier pilings offsets colonization of cryptogenic and non-indigenous species in a Chilean port.

Examining the effects of disturbances within marine urban communities can shed light on their assembly rules and invasion processes. The effects of physical disturbance, through the removal of dominant native habitat-builders, were investigated in the recolonization of disturbed patches and colonization of plates on pier pilings, in a Chilean port. On pilings, disturbance substantially affected community structure after 3 months, although it slowly converged across treatments after 10 months. On plates, cryptogenic and non-indigenous species richness increased with removal severity, which was not observed in natives. Opportunistic taxa took advantage of colonizing at an early successional stage, illustrating a competition-colonization trade-off, although indirect effects might be at play (e.g. trophic competition or selective predation). Recovery of the habitat-builders then occurred at the expense of cryptogenic and non-indigenous taxa. Whether natives could continue winning against increasing propagule and colonization pressures in marine urban habitats deserves further attention. The interactions between disturbance and biological invasions herein experimentally shown in situ contribute to our understanding of multiple changes imposed by marine urbanization in a growing propagule transport network.

Gene flow in the Antarctic bivalve Aequiyoldia eightsii (Jay, 1839) suggests a role for the Antarctic Peninsula Coastal Current in larval dispersal.

The Antarctic Circumpolar Current (ACC) dominates the open-ocean circulation of the Southern Ocean, and both isolates and connects the Southern Ocean biodiversity. However, the impact on biological processes of other Southern Ocean currents is less clear. Adjacent to the West Antarctic Peninsula (WAP), the ACC flows offshore in a northeastward direction, whereas the Antarctic Peninsula Coastal Current (APCC) follows a complex circulation pattern along the coast, with topographically influenced deflections depending on the area. Using genomic data, we estimated genetic structure and migration rates between populations of the benthic bivalve Aequiyoldia eightsii from the shallows of southern South America and the WAP to test the role of the ACC and the APCC in its dispersal. We found strong genetic structure across the ACC (between southern South America and Antarctica) and moderate structure between populations of the WAP. Migration rates along the WAP were consistent with the APCC being important for species dispersal. Along with supporting current knowledge about ocean circulation models at the WAP, migration from the tip of the Antarctic Peninsula to the Bellingshausen Sea highlights the complexities of Southern Ocean circulation. This study provides novel biological evidence of a role of the APCC as a driver of species dispersal and highlights the power of genomic data for aiding in the understanding of the influence of complex oceanographic processes in shaping the population structure of marine species.

After a catastrophe, a little bit of sex is better than nothing: Genetic consequences of a major earthquake on asexual and sexual populations.

Catastrophic events can have profound effects on the demography of a population and consequently on genetic diversity. The dynamics of postcatastrophic recovery and the role of sexual versus asexual reproduction in buffering the effects of massive perturbations remain poorly understood, in part because the opportunity to document genetic diversity before and after such events is rare. Six natural (purely sexual) and seven cultivated (mainly clonal due to farming practices) populations of the red alga Agarophyton chilense were surveyed along the Chilean coast before, in the days after and 2 years after the 8.8 magnitude earthquake in 2010. The genetic diversity of sexual populations appeared sensitive to this massive perturbation, notably through the loss of rare alleles immediately after the earthquake. By 2012, the levels of diversity returned to those observed before the catastrophe, probably due to migration. In contrast, enhanced rates of clonality in cultivated populations conferred a surprising ability to buffer the instantaneous loss of diversity. After the earthquake, farmers increased the already high rate of clonality to maintain the few surviving beds, but most of them collapsed rapidly. Contrasting fates between sexual and clonal populations suggest that betting on strict clonality to sustain production is risky, probably because this extreme strategy hampered adaptation to the brutal environmental perturbation induced by the catastrophe.

The influence of glacial melt and retreat on the nutritional condition of the bivalve Nuculana inaequisculpta (Protobranchia: Nuculanidae) in the West Antarctic Peninsula.

Due to climate change, numerous ice bodies have been lost in the West Antarctic Peninsula (WAP). As a consequence, deglaciation is expected to impact the marine environment and its biota at physiological and ecosystem levels. Nuculana inaequisculpta is a marine bivalve widely distributed around Antarctica that plays an important role for ecosystem functioning. Considering that N. inaequisculpta inhabits coastal areas under effect of glacial melt and retreat, impacts on its nutritional condition are expected due to alterations on its physiology and food availability. To test this hypothesis, biochemical composition (lipids, proteins, and fatty acids) and energy content were measured in individuals of N. inaequisculpta collected in a fjord at different distances to the retreating glacier in the WAP. Oceanographic parameters of the top and bottom-water layers (temperature, salinity, dissolved oxygen, and chlorophyll-a) were measured to investigate how the environment changes along the fjord. Results showed that surface oceanographic parameters displayed a lower temperature and dissolved oxygen, but a higher salinity and chlorophyll-a content at nearest compared to farthest sites to the glacier. In contrast, a lower temperature and chlorophyll-a, and a higher salinity and dissolved oxygen was measured in the bottom-water layer toward the glacier. N. inaequisculpta had a higher amount of lipids (17.42 ± 3.24 vs. 12.16 ± 3.46%), protein (24.34 ± 6.12 vs. 21.05 ± 2.46%) and energy content (50.57 ± 6.97 J vs. 39.14 ± 5.80 J) in the farthest compared to the nearest site to the glacier. No differences were found in total fatty acids among all sites. It seems likely that lower individual fitness related to proximity to the glacier would not be related to nutritional quality of sediment food, but rather to food quantity.

Short communication: Characterization of the expression of microRNAs in the poecilogonous polychaete B. wellingtonensis.

Poecilogony is a type of reproduction in which a species produces different types of larvae. Boccardia wellingtonensis, is a poecilogonous polychaete with females producing planktotrophic and adelphophagic larvae, in addition to nurse eggs, in the same capsule that differ in feeding behavior. It is still unclear why planktotrophs do not feed on nurse eggs during the intracapsular development and arrest its growth, while adelphophagic larvae consume nurse eggs and planktotrophic larvae inside the capsule, hatching as advance larvae or as juveniles. Here we characterized the expression of selected miRNAs from these two types of larvae and from adults in order to begin to understand the molecular mechanisms that regulate expression in this type of poecilogony. Results showed that adults and pre-hatching adelphophagic larvae have high levels of expression of miR-125, miR-87a and let-7, while adelphophages at early developmental stage had low levels of expression of miR-87b. Planktotrophic larvae showed low expression level of let-7. This work represents the first step in understanding the role of miRNAs in the development of different larval types in a poecilogonous species. We also propose to B. wellingtonensis as an interesting biological model to study the evolution of larval modes and reproductive strategies of marine invertebrates.

Post-Disturbance Genetic Changes: The Impact of the 2010 Mega-Earthquake and Tsunami on Chilean Sandy Beach Fauna.

Earthquake/tsunamis can have profound impacts on species and their genetic patterns. It is expected that the magnitude of this impact might depend on the species and the time since the disturbance occurs, nevertheless these assumptions remain mostly unexplored. Here we studied the genetic responses of the crustacean species Emerita analoga, Excirolana hirsuticauda, and Orchestoidea tuberculata to the 27F mega-earthquake/tsunami that occurred in Chile in February 2010. mtDNA sequence analyses revealed a lower haplotype diversity for E. analoga and E. hirsuticauda in impacted areas one month after the 27F, and the opposite for O. tuberculata. Three years after the 27F we observed a recovery in the genetic diversity of E. analoga and E. hirsuticauda and decrease in the genetic diversity in O. tuberculata in 2/3 of sampled areas. Emerita analoga displayed decrease of genetic differentiation and increase in gene flow explained by long-range population expansion. The other two species revealed slight increase in the number of genetic groups, little change in gene flow and no signal of population expansion associated to adult survival, rapid colonization, and capacity to burrow in the sand. Our results reveal that species response to a same disturbance event could be extremely diverse and depending on life-history traits and the magnitude of the effect.

Spread of the non-native anemone Anemonia alicemartinae Häussermann & Försterra, 2001 along the Humboldt-current large marine ecosystem: an ecological niche model approach.

The geographical expansion of invasive species depends mainly on its dispersal potential, and the abiotic and biotic factors affecting it. Knowing the invasive dynamic of non-native species, as well as its behavior at different natural or anthropogenic scenarios, is fundamental for planning conservation management policies and control plans. The invasive sea anemone Anemonia alicemartinae in habits from the north (18°S) to the south-central (36°S) coast of Chile and its distribution range has expanded by approximately 1,928 km in the last 50 years. Previous works have proposed that human-mediated southward transport associated with regional-scale maritime activities could explain its rapid spread. To evaluate this hypothesis, we used ecological niche models (ENM) to evaluate the potential colonization of the southernmost area of South America. Additionally, we conducted a post hoc analysis to evaluate the relationship between the prediction of the ENM and human activity measured as the number of landings of ships in ports. The models were built based on presence records of A. alicemartinae, and oceanographic variables. Results showed that sea surface salinity and annual sea surface temperature (variance) are the best predictor variables to explain the distribution of A. alicemartinae. There was a positive and significant relationship between the geographical distribution of the sea anemone predicted by the ENM and the number of landings, as a proxy of anthropogenic activity. The most susceptible areas to invasion were those that showed the highest variability in both oceanographic predictors. These areas included the Biobío region, Chiloé´s inland sea, Aysén, and Chacabuco regions, which together comprise two biogeographical provinces. These results sustain the proposed hypothesis and, overall, the results suggest that along with the characteristics of the life history of A. alicemartinae, oceanographic conditions and maritime transport as vector contribute to the southern range expansion of this invasive cryptogenic species in the Humboldt-current large marine ecosystem.

Contrasting Metatrochal Behavior of Mollusc and Annelid Larvae and the Regulation of Feeding While Swimming.

Molluscan veliger larvae and some annelid larvae capture particulate food between a preoral prototrochal band of long cilia that create a current for both swimming and feeding and a postoral metatrochal band of shorter cilia that beat toward the prototroch. Larvae encountering satiating or noxious particles must somehow swim without capturing particles or else reject large numbers of captured particles. Because high rates of particle capture are inferred to depend on the beat of both ciliary bands, arrest of the metatroch could be one way to swim while reducing captures. Larvae in eight families of annelids arrest metatrochal cilia frequently during prototrochal beat, often over a large part of the metatrochal band and with the arrested cilia aligned near the beginning of the effective stroke. In contrast, metatrochs of veligers of gastropods and bivalves rarely arrested while the prototroch beat, and those arrests were more localized and variable in position. This difference in metatrochal arrest was unexpected under hypotheses of either a single origin of this feeding mechanism or multiple origins within each phylum. Although different in metatrochal arrests, larvae of both phyla can separate swimming from feeding while both prototroch and metatroch beat. One hypothesis explaining low rates of capture per encounter, without metatrochal arrest, is a change in adhesion of prototrochal cilia with algae. In a few observations, part of the velar edge was retained within the veliger's shell so that exposed prototrochal cilia contributed to swimming while the adjacent metatroch and food groove were sequestered.

Role of the larval feeding morphology and digestive enzyme activity in the early development of the polychaete Boccardia wellingtonensis.

In marine invertebrates, the modes of development at early stages are related to the type and capacity of larval feeding to achieve growth. Therefore, studying the factors that determine larval feeding strategies can help to understand the diversity of life histories and evolution of marine invertebrates. The polychaete Boccardia wellingtonensis is a poecilogonous species that encapsulates and incubates its offspring. This species produces two types of larvae: (1) larvae that do not feed within the capsule and hatch as planktotrophic larvae (indirect development), and (2) adelphophagic larvae that feed on nurse eggs and other larvae inside the capsule to hatch as advanced larvae or juveniles (direct development). Otherwise, the larval types are indistinguishable at the same stage of development. The non-apparent morphological differences between both types of larvae suggest that other factors are influencing their feeding behavior. This work studied the potential role of the activity of 19 digestive enzymes on the different feeding capacities of planktotrophic and adelphophagic larvae of B. wellingtonensis. Also, differences in larval feeding structures and the larval capacity to feed from intracapsular fluid were evaluated by electron and fluorescence microscopy. Results showed that both types of larvae present similar feeding structures and had the capacity to ingest intracapsular fluid protein. Adelphophagic larvae showed overall the highest activities of digestive enzymes. Significant differences between larval types were observed in nine enzymes related to the use of internal and external nutritional sources. Given that larval feeding is closely related to larval development in species with encapsulation, this work supports that the study of the digestive enzymatic machinery of larvae may contribute to understanding the evolution of developmental modes.

Non-indigenous species contribute equally to biofouling communities in international vs local ports in the Biobío region, Chile.

Growing coastal urbanization together with the intensification of maritime traffic are major processes explaining the increasing rate of biological introductions in marine environments. To investigate the link between international maritime traffic and the establishment of non-indigenous species (NIS) in coastal areas, biofouling communities in three international and three nearby local ports along 100 km of coastline in south-central Chile were compared using settlement panels and rapid assessment surveys. A larger number of NIS was observed in international ports, as expected in these 'invasion hubs'. However, despite a few environmental differences between international and local ports, the two port categories did not display significant differences regarding NIS establishment and contribution to community structure over the studied period (1.5 years). In international ports, the free space could be a limiting factor for NIS establishment. The results also suggest that local ports should be considered in NIS surveillance programs in Chile.

The alien slipper limpet Crepipatella dilatata (Lamarck, 1819) in northern Spain: A multidisciplinary approach to its taxonomic identification and invasive biology.

The slipper limpet Crepipatella dilatata, native to Chile and Argentina, was introduced in Spain in 2005. The species was thought to inhabit the region of Rias Bajas, yet recently, putative C. dilatata populations have been documented on the coast of north-central Spain and in the Ebro Delta of the Spanish Mediterranean. Here we undertook a multidisciplinary approach to study the invasion biology of this species. Specifically, two geographically distant populations, one being a successfully established population from O Grove and the other a declining population from Gijon, were studied over the course of four years. Analyses of morphological and developmental traits as well as genetic information confirmed the presence of C. dilatata in these sites. The results revealed polymorphism in anatomical traits and shell shape. Shell shape polymorphism was unevenly distributed among sites and among sexes. Males were monomorphic, while females were polymorphic. Of the female morphotypes encountered, one was absent in the declining population from Gijón. Size at first female maturation and female size were greater in the declining population than in the established population. Reproductive success varied seasonally but not spatially among populations. In the established population, gregariousness was significantly greater; the size when sex changes was found to be plastic and socially controlled. The sex ratio of the declining population was female biased while in the established population the sex ratio changed during the study period from being balanced to being female biased. This change in sex ratio was probably due to higher male mortality. Molecular analyses pointed to the localities of Corral Bay in southern Chile and Puerto Madryn in southern Argentina as potential population sources. The intercontinental import of fresh mussels cultivated in Chilean farms is a likely source of this mussel in Spain. Comparison with available data of native populations of C. dilatata strongly indicate that ecophenotypic plasticity, socially controlled sex change, high gregariousness, increased nurse egg supply to viable larvae during the encapsulated developmental period, later maturation and larger female sizes altogether enhance establishment success of this non-indigenous species. Human-mediated factors like the intraregional mussel trade and transplantation are also likely secondary dispersal mechanisms favouring the spread of this organism.

The identification of sympatric cryptic free-living nematode species in the Antarctic intertidal.

The diversity of free-living nematodes in the beaches of two Antarctic islands, King George and Deception islands was investigated. We used morphological and molecular (LSU, and two fragments of SSU sequences) approaches to evaluate 236 nematodes. Specimens were assigned to at least genera using morphology and were assessed for the presence of cryptic speciation. The following genera were identified: Halomonhystera, Litoditis, Enoploides, Chromadorita, Theristus, Oncholaimus, Viscosia, Gammanema, Bathylaimus, Choanolaimus, and Paracanthonchus; along with specimens from the families Anticomidae and Linhomoeidae. Cryptic speciation was identified within the genera Halomonhystera and Litoditis. All of the cryptic species identified live sympatrically. The two cryptic species of Halomonhystera exhibited no significant morphological differences. However, Litoditis species 2 was significantly larger than Litoditis species 1. The utility of molecular data in confirming the identifications of some of the morphologically more challenging families of nematodes was demonstrated. In terms of which molecular sequences to use for the identification of free-living nematodes, the SSU sequences were more variable than the LSU sequences, and thus provided more resolution in the identification of cryptic speciation. Finally, despite the considerable amount of time and effort required to put together genetic and morphological data, the resulting advance in our understanding of diversity and ecology of free-living marine nematodes, makes that effort worthwhile.

Presence of the tunicate Asterocarpa humilis on ship hulls and aquaculture facilities in the coast of the Biobío Region, south central Chile.

Non-native ascidians are important members of the fouling community associated with artificial substrata and man-made structures. Being efficient fouling species, they are easily spread by human-mediated transports (e.g., with aquaculture trade and maritime transports). This is exemplified by the ascidian Asterocarpa humilis which displays a wide distribution in the Southern Hemisphere and has been recently reported in the Northern Hemisphere (NW Europe). In continental Chile, its first report dates back from 2000 for the locality of Antofagasta (23°S). Although there was no evidence about the vectors of introduction and spread, nor the source, some authors suggested maritime transport by ship hulls and aquaculture devices as putative introduction pathways and vectors. In the present study, we report for the first time the presence of A. humilis on the hull of an international ship in a commercial port in Concepción bay (36°S), south central Chile. We also found one individual associated to a seashell farm, 70 km far from Concepción bay. Further individuals were subsequently identified within Concepción bay: one juvenile settled upon international harbor pilings and a dozen individuals along aquaculture seashell longlines. For the first specimens sampled, species identification was ascertained using both morphological criteria and molecular barcoding, using the mitochondrial gene cytochrome c oxidase subunit I (COI) and a nuclear gene (ribosomal RNA 18S). The nuclear 18S gene and the mitochondrial gene COI clearly assigned the specimens to A. humilis, confirming our morphological identification. Two haplotypes were obtained with COI corresponding to haplotypes previously obtained with European and Northern Chilean specimens. The present study thus reports for the first time the presence of A. humilis in the Araucanian ecoregion, documenting the apparent expansion of this non-native tunicate in Chile over 2,000 km, spanning over three ecoregions. In addition we reveal the potential implication of the international maritime transport as a vector of spread of this species along the Eastern Pacific coast, and the putative role of aquaculture facilities in promoting local establishments of non-native tunicates.