Dr. Brante-Ramírez, Antonio

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 How to cite this article Pinochet J, Rivera R, Neill PE, Brante A, Hernández CE. 2019. Spread of the non-native anemone Anemonia alicemartinae Häussermann & Försterra, 2001 along the Humboldt-current large marine ecosystem: an ecological niche model approach. maritime transport as vector contribute to the southern range expansion of this invasive cryptogenic species in the Humboldt-current large marine ecosystem.

The distribution range expansion of species mediated by natural or anthropic mechanisms is one of the main causes of changes in biodiversity patterns. Anemonia alicemartinae is a cryptogenic species found along the coasts of the Southeast Pacific Ocean. This species has expanded its range by >1900 km along the Chilean coast throughout the last 50 yr. A. alicemartinae cohabits with the native anemone Phymactis papillosa in the low intertidal zone, and given the limited mobility of both species, limited space could encourage aggressive behavior between them. P. papillosa shows different color phenotypes, and, as in other anemone species, color is associated with its level of aggressiveness. Here, we evaluated the aggressive behavior of A. alicemartinae on 2 color morphotypes of P. papillosa. Also, intraspecific agonistic interactions were considered in individuals of A. alicemartinae from 2 localities. Four experiments were conducted: (1) individual vs. individual, (2) individual vs. group forming a frontal line, (3) individual vs. group surrounding the individual and (4) group vs. group. Results showed A. alicemartinae to be a weak competitor against P. papillosa. The cryptogenic species lost more contests with green P. papillosa than with the red phenotype. Few aggressive interactions between conspecifics of A. alicemartinae were found. Results suggest that the expansion success of A. alicemartinae could be explained by alternative strategies, such as escape behavior, asexual reproduction and high dispersal potential.

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 diferentiation and increase in gene fow explained by long-range population expansion. The other two species revealed slight increase in the number of genetic groups, little change in gene fow 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 efect.

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.

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 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.

Studying the mating system of obligate aquatic organisms that inhabit river ecosystems is important for understanding its evolution as well as the role of biological and environmental factors in modulating population dynamics and species distributional patterns. Here, we studied the reproductive strategy of the Chilean endemic freshwater snail, Chilina dombeiana,in the BiobõÂo River, one of the largest rivers in Chile. This species has a low potential for dispersal given the absence of a free-swimming larval stage (benthic larval development)and given that adults have a low capacity for mobility. We hypothesized that: 1. Females would mate with different males (polyandry resulting in intrabrood multiple paternity, 2. Individuals from closer sites would be more related than individuals from distant sites, and 3. Male parental contributions would be unevenly distributed within broods. Individuals from three different sites were sampled along the river: upper, mid, and river mouth. In the laboratory, hatching juveniles from a total of 15 broods were collected for paternity analyses. We used microsatellite markers and the programs GERUD and COLONY to determine whether multiple paternity exists and to estimate the contribution of different males to the brood. We found that multiple paternity was very common at all of the sites analyzed with as many as 8 males fertilizing a single female and a mean of 4.2 fathers per brood estimated by COLONY. Sire contribution was skewed to particular males in several broods. In addition, overall relatedness among broods for the three sites ranged from 0.17 to 0.45 with evidence of many half-siblings. Relatedness differed among the three sites. Particularly in upstream sites or in anthropogenically disturbed populations, the high levels of multiple paternity observed in C. dombeiana may be an efficient strategy to avoid inbreeding and prevent the loss of genetic diversity within populations.

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_x000E_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_x000E_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.

Protandric species switch sex during their lifetime. According to theory, the time (body size) at which sex change occurs is determined by the reproductive success of individuals affected by social interactions as well as by post-copulatory factors. Experimental evidence is biased to few social systems making the exploration of general patterns difficult. We used the protandric marine gastropod Crepidula coquimbensis that partakes in intrabrood sibling cannibalism to test the following hypotheses: 1. Male-male competition for access to females and sibling cannibalism determine male reproductive success; 2. Males with greater access to females and with higher reproductive success will have reduced growth rates and will delay sex change. Artificial aggregations with different social structures were constructed and male reproductive success was estimated by paternity analysis. The results supported our expectations showing that male competitive ability for access to the female, time spent by males in the copulatory position, and sibling cannibalism affect reproductive success and influence time to sex change, with less successful males hastening sex change. Also, males that spent more time in the copulatory position had reduced growth rates. Comparing these results with those reported for other sequential hermaphrodites provides evidence supporting general patterns of sex change in nature.

The study of reproduction and mating beahavior constitutes a main issue in biology, ecology and evolution, given its relation with fitness traits. Here, we developed a simple microsatellite multiple assay to evaluate the mating strategy and male reproductive success of the marine gastropod Concholepas concholepas (Bruguie `re, 1789), an important fishery resource and a key predator species of Chilean rocky shore communities. Concholepas concholepas is a dioecious species with internal fertilization, encapsulation, and long larval phase. In laboratory, adult males and females were cultivated in tanks, and 37 larvae from 5 different clutches were genotyped to run paternity analyses using seven microsatellite loci. Results showed that promiscuity is a common mating behavior in C. concholepas displaying an exceptionally high level of multipaternity and males participating as fathers in clutches from more than one female. This microsatellite multiple assay helped to improve our understanding of the reproductive beahavior of this ecological key species with high economic importance.