Does the loss of diadromy imply the loss of salinity tolerance? A gene expression study with replicate nondiadromous populations of Galaxias maculatus.

The recurrent colonization of freshwater habitats and subsequent loss of diadromy is a major ecological transition that has been reported in many ancestrally diadromous fishes. Such residency is often accompanied by a loss of tolerance to seawater. The amphidromous Galaxias maculatus has repeatedly colonized freshwater streams with evidence that freshwater-resident populations exhibit stark differences in their tolerance to higher salinities. Here, we used transcriptomics to gain insight into the mechanisms contributing to reduced tolerance to higher salinities in freshwater resident populations. We conducted an acute salinity challenge (0 ppt to 23-25 ppt) and measured osmoregulatory ability (muscle water content) over 48 h in three populations: diadromous, saltwater intolerant resident (Toltén), and saltwater tolerant resident (Valdivia). RNA sequencing of the gills identified genes that were differentially expressed in association with the salinity change and associated with the loss of saltwater tolerance in the Toltén population. Key genes associated with saltwater acclimation were characterized in diadromous G. maculatus individuals, some of which were also expressed in the saltwater tolerant resident population (Valdivia). We found that some of these "saltwater acclimation" genes, including the cystic fibrosis transmembrane conductance regulator gene (CFTR), were not significantly upregulated in the saltwater intolerant resident population (Toltén), suggesting a potential mechanism for the loss of tolerance to higher salinities. As the suite of differentially expressed genes in the diadromous-resident comparison differed between freshwater populations, we hypothesize that diadromy loss results in unique evolutionary trajectories due to drift, so the loss of diadromy does not necessarily lead to a loss in upper salinity tolerance.

La colonización recurrente de hábitats de agua dulce y la subsecuente pérdida de diadromía es una transición ecológica importante que ha sido reportada en varias especies de peces con ancestros diádromos. Esta residencia está acompañada frecuentemente por la pérdida de tolerancia a ambientes de agua salada. Galaxias maculatus, especie anfídroma, ha colonizado ríos repetidamente y existe evidencia que las poblaciones residentes presentan diferencias respecto a la tolerancia al agua salada. En este estudio, usamos transcriptómica para dilucidar los mecanismos que contribuyen a la reducida tolerancia a altas salinidades en las poblaciones residentes de agua dulce. Realizamos un desafío agudo de salinidad (0 ppt a 23-2 ppt) y medimos la habilidad osmoreguladora (contenido de agua en músculo) por 48 horas en individuos de tres poblaciones: una diádroma, una intolerante a agua salada (Toltén) y una tolerante a agua salada (Valdivia). Con el secuenciamiento de ARN de las branquias identificamos los genes expresados diferencialmente al cambio de salinidad y cuales están asociados a la pérdida de tolerancia a agua salada en la población de Toltén. Genes claves asociados a la aclimatación al agua salada fueron caracterizados en individuos diádromos, algunos de ellos también se expresaron en la población residente tolerante al agua salada (Valdivia). Sin embargo, algunos genes involucrados en la aclimatación al agua salada, incluyendo el gen regulador de la conductancia transmembrana de la fibrosis quística (CFTR), no se diferenciaron significativamente en la población residente intolerante al agua salada (Toltén), sugiriendo un mecanismo potencial de la pérdida de tolerancia a ambientes con salinidad elevada. Como el conjunto de genes expresados difiere entre las dos poblaciones residentes al compararse con la población diádroma, hipotetizamos que la pérdida de diadromía resulta en trayectorias evolutivas únicas debido a deriva génica, por lo que la pérdida de la diadromía no necesariamente conlleva a la pérdida de la tolerancia a aguas saladas.

Hydrological connectivity drives longitudinal movement of endangered endemic Chilean darter Percilia irwini (Eigenmann, 1927).

Movement is a fundamental aspect of fish ecology, and it therefore represents an important trait to monitor for the management and conservation of fish populations. This is especially true for small benthic fish, as they often inhabit part of the catchment where their movement may be restricted by alterations to river connectivity due to human activity. Still, the movement of these small benthic fish remains poorly understood, partly because of their small size and their cryptic nature. This applies to Percilia irwini, an endangered small darter native to the south-central region of Chile. Its habitat has been affected by the presence of large hydroelectric dams and is currently threatened by the construction of several others. In this study, the authors investigated movement patterns of P. irwini from populations inhabiting different parts of the Biobío catchment, with different levels of connectivity due to natural and/or human-induced features. The authors combined chronological clustering with random forest classification to reconstruct lifelong movements from multi-elemental otolith microchemistry transects. The majority of the movements detected occurred in an undisturbed part of the catchment. These were directional upstream movements occurring between capture sites from the lower and the middle reaches of the river, representing a distance of nearly 30 km, a distance much larger than previously thought. Nonetheless, in the part of the catchment where connectivity was affected by human activity, no such movements were identified. This study shows that connectivity alteration could impede naturally occurring movement and further threaten the resilience of populations of P. irwini. Furthermore, the results presented are used to discuss advantages and disadvantages of microchemistry analysis for studying movement of small benthic fish.

Multiple drainage reversal episodes and glacial refugia in a Patagonian fish revealed by sequenced microsatellites.

The rise of the southern Andes and the Quaternary glacial cycles influenced the landscape of Patagonia, affecting the phylogeographic and biogeographic patterns of its flora and fauna. Here, we examine the phylogeography of the freshwater fish, Percichthys trucha, using 53 sequenced microsatellite DNA markers. Fish (n= 835) were collected from 16 river systems (46 locations) spanning the species range on both sides of the Andes. Eleven watersheds drain to the Pacific, five of which are trans-Andean (headwaters east of Andes). The remaining five drainages empty into the Atlantic. Three analytical approaches (neighbour-joining tree, hierarchical AMOVAs, Structure) revealed evidence of historic drainage reversals: fish from four of the five trans-Andean systems (Puelo, Futalaufquen/Yelcho, Baker, Pascua) exhibited greater genetic similarity with Atlantic draining systems than with Pacific systems with headwaters west of Andes. Present-day drainage (Pacific versus Atlantic) explained only 5% of total genetic variance, while ancestral drainage explained nearly 27% of total variance. Thus, the phylogeographic structure of P. trucha is consistent with episodes of drainage reversal in multiple systems and suggests a major role for deglaciation in the genetic and indeed the geographical distribution of P. trucha in Patagonia. The study emphasizes the significant role of historical processes in the current pattern of genetic diversity and differentiation in a fish from a southern temperate region.

Genomic basis of the loss of diadromy in Galaxias maculatus: Insights from reciprocal transplant experiments.

Diadromy is known for having major effects on the distribution and richness of aquatic species, and so does its loss. The loss of diadromy has led to the diversification of many species, yet research focusing on understanding its molecular basis and consequences are limited. This is particularly true for amphidromous species despite being the most abundant group of diadromous species. Galaxias maculatus, an amphidromous species and one of the most widely distributed fishes in the Southern Hemisphere, exhibits many instances of nonmigratory or resident populations. The existence of naturally replicated resident populations in Patagonia can serve as an ideal system for the study of the mechanisms that lead to the loss of the diadromy and its ecological and evolutionary consequences. Here, we studied two adjacent river systems in which resident populations are genetically differentiated yet derived from the same diadromous population. By combining a reciprocal transplant experiment with genomic data, we showed that the two resident populations followed different evolutionary pathways by exhibiting a differential response in their capacity to survive in salt water. While one resident population was able to survive salt water, the other was not. Genomic analyses provided insights into the genes that distinguished (a) migratory from nonmigratory populations; (b) populations that can vs those that cannot survive a saltwater environment; and (c) between these resident populations. This study demonstrates that the loss of diadromy can be achieved by different pathways and that environmental (selection) and random (genetic drift) forces shape this dynamic evolutionary process.

Biogeography, habitat transitions and hybridization in a radiation of South American silverside fishes revealed by mitochondrial and genomic RAD data.

Rivers and lake systems in the southern cone of South America have been widely influenced by historical glaciations, carrying important implications for the evolution of aquatic organisms, including prompting transitions between marine and freshwater habitats and by triggering hybridization among incipient species via waterway connectivity and stream capture events. Silverside fishes (Odontesthes) in the region comprise a radiation of 19 marine and freshwater species that have been hypothesized on the basis of morphological or mitochondrial DNA data to have either transitioned repeatedly into continental waters from the sea or colonized marine habitats following freshwater diversification. New double digest restriction-site associated DNA data presented here provide a robust framework to investigate the biogeographical history of and habitat transitions in Odontesthes. We show that Odontesthes silversides originally diversified in the Pacific but independently colonized the Atlantic three times, producing three independent marine-to-freshwater transitions. Our results also indicate recent introgression of marine mitochondrial haplotypes into two freshwater clades, with more recurring instances of hybridization among Atlantic- versus Pacific-slope species. In Pacific freshwater drainages, hybridization with a marine species appears to be geographically isolated and may be related to glaciation events. Substantial structural differences of estuarine gradients between these two geographical areas may have influenced the frequency, intensity and evolutionary effects of hybridization events.

Invasive species and postglacial colonization: their effects on the genetic diversity of a Patagonian fish.

The present distribution of Patagonian species is the result of a complex history involving Quaternary refugial populations, Holocene range expansions and demographic changes occurring during the Anthropocene. Invasive salmonids were introduced in Patagonia during the last century, occupying most rivers and lakes, preying on and competing with native species, including the fish Galaxias platei. Here, we used G. platei as a case study to understand how long-term (i.e. population differentiation during the Holocene) and short-term historical processes (salmonid introductions) affect genetic diversity. Using a suite of microsatellite markers, we found that the number of alleles is negatively correlated with the presence of salmonids (short-term processes), with G. platei populations from lakes with salmonids exhibiting significantly lower genetic diversity than populations from lakes without salmonids. Simulations (100 years backwards) showed that this difference in genetic diversity can be explained by a 99% reduction in population size. Allelic richness and observed heterozygosities were also negatively correlated with the presence of salmonids, but also positively correlated with long-term processes linked to Quaternary glaciations. Our results show how different genetic parameters can help identify processes taking place at different scales and their importance in terms of conservation.

The effects of diadromy and its loss on genomic divergence: The case of amphidromous Galaxias maculatus populations.

Understanding the evolutionary mechanisms that affect the genetic divergence between diadromous and resident populations across heterogeneous environments is a challenging task. While diadromy may promote gene flow leading to a lack of genetic differentiation among populations, resident populations tend to be affected by local adaptation and/or plasticity. Studies on these effects on genomic divergence in nonmodel amphidromous species are scarce. Galaxias maculatus, one of the most widespread fish species in the Southern Hemisphere, exhibits two life histories, an ancestral diadromous, specifically, amphidromous form, and a derived freshwater resident form. We examined the genetic diversity and divergence among 20 estuarine and resident populations across the Chilean distribution of G. maculatus and assessed the extent to which selection is involved in the differentiation among resident populations. We obtained nearly 4,400 SNP markers using a RADcap approach for 224 individuals. As expected, collections from estuarine locations typically consist of diadromous individuals. Diadromous populations are highly differentiated from their resident counterparts by both neutral and putative adaptive markers. While diadromous populations exhibit high gene flow and lack site fidelity, resident populations appear to be the product of different colonization events with relatively low genetic diversity and varying levels of gene flow. In particular, the northernmost resident populations were clearly genetically distinct and reproductively isolated from each other suggesting local adaptation. Our study provides insights into the role of life history differences in the maintenance of genetic diversity and the importance of genetic divergence in species evolution.

Past, present, and future of a freshwater fish metapopulation in a threatened landscape.

It is well documented that hydropower plants can affect the dynamics of fish populations through landscape alterations and the creation of new barriers. Less emphasis has been placed on the examination of the genetic consequences for fish populations of the construction of dams. The relatively few studies that focus on genetics often do not consider colonization history and even fewer tend to use this information for conservation purposes. As a case study, we used a 3-pronged approach to study the influence of historical processes, contemporary landscape features, and potential future anthropogenic changes in landscape on the genetic diversity of a fish metapopulation. Our goal was to identify the metapopulation's main attributes, detect priority areas for conservation, and assess the consequences of the construction of hydropower plants for the persistence of the metapopulation. We used microsatellite markers and coalescent approaches to examine historical colonization processes, traditional population genetics, and simulations of future populations under alternate scenarios of population size reduction and gene flow. Historical gene flow appeared to have declined relatively recently and contemporary populations appeared highly susceptible to changes in landscape. Gene flow is critical for population persistence. We found that hydropower plants could lead to a rapid reduction in number of alleles and to population extirpation 50-80 years after their construction. More generally, our 3-pronged approach for the analyses of empirical genetic data can provide policy makers with information on the potential impacts of landscape changes and thus lead to more robust conservation efforts.

Echoes of a distant time: effects of historical processes on contemporary genetic patterns in Galaxias platei in Patagonia.

Interpreting the genetic structure of a metapopulation as the outcome of gene flow over a variety of timescales is essential for the proper understanding of how changes in landscape affect biological connectivity. Here we contrast historical and contemporary connectivity in two metapopulations of the freshwater fish Galaxias platei in northern and southernmost Patagonia where paleolakes existed during the Holocene and Pleistocene, respectively. Contemporary gene flow was mostly high and asymmetrical in the northern system while extremely reduced in the southernmost system. Historical migration patterns were high and symmetric in the northern system and high and largely asymmetric in the southern system. Both systems showed a moderate structure with a clear pattern of isolation by distance (IBD). Effective population sizes were smaller in populations with low contemporary gene flow. An approximate Bayesian computation (ABC) approach suggests a late Holocene colonization of the lakes in the northern system and recent divergence of the populations from refugial populations from east and west of the Andes. For the southern system, the ABC approach reveals that some of the extant G. platei populations most likely derive from an ancestral population inhabiting a large Pleistocene paleolake while the rest derive from a higher-altitude lake. Our results suggest that neither historical nor contemporary processes individually fully explain the observed structure and geneflow patterns and both are necessary for a proper understanding of the factors that affect diversity and its distribution. Our study highlights the importance of a temporal perspective on connectivity to analyse the diversity of spatially complex metapopulations.

Phylogeography of the catfish Hatcheria macraei reveals a negligible role of drainage divides in structuring populations.

Southern South America provides a set of unusual geographic features that make it particularly interesting for studying phylogeography. The Andes Mountains run along a north-to-south axis and act as a barrier to gene flow for much of the biota of this region, with southern portions experiencing extensive historical glaciation. Geological data reveal a series of drainage reversals, shifting from Pacific Ocean outlets to Atlantic Ocean outlets because of glacier formation that dammed and reversed rivers. Once glaciers melted around 13 000 years ago, drainages returned to the Pacific Ocean. This geologic history predicts that aquatic organisms in Pacific rivers should have their closest relationships to their counterparts in Atlantic rivers immediately to their east. We tested this prediction in the trichomycterid catfish Hatcheria macraei from 38 locations using the mitochondrial cytochrome b gene. Our results show that most populations found in Pacific rivers were closely related to fish found in the adjacent Atlantic draining Río Chubut. Surprisingly, one documented drainage reversal (from Río Deseado into Río Baker) did not result in movement of H. macraei. Overall, we found the lowest levels of genetic structure between most Pacific rivers that are adjacent to the Atlantic draining Río Chubut. We also found low levels of population structuring among three of four contemporary river basins that drain to the Atlantic Ocean. Our findings suggest that drainage basin boundaries have historically not played an important long-term role in structuring between nine of 11 drainages, an unusual finding in freshwater biogeography.

Polybrominated diphenyl ether levels in wild and farmed Chilean salmon and preliminary flow data for commercial transport.

This pilot study documented the occurrence and levels of brominated flame retardants in the tissues of farmed and wild salmon in southern Chile. Samples of Coho salmon and rainbow trout were obtained from fish farms, rivers and lakes in the Patagonia in Aysen Region, Chile. The samples were analyzed by Gas Chromatography Negative Chemical Ionization Mass Spectrometry for the different polybrominated diphenyl ether (PBDE) congeners. Contaminants were observed in all the samples, and the congeners BDE 17, 28, 47 and 66 were observed in all both farmed and wild samples. The concentrations were higher in the farmed Coho salmon, presenting significant differences with wild salmon. The levels reached 182 pg/g wet weight (ww) vs. 120 ww. In the case of the rainbow trout, the concentrations were lower, although the congener profile was quite similar. The levels reached an average of 100 pg/g ww in the farmed fish versus 110 pg/g ww in wild fish, and no significant difference was observed between the species. In both species, the congener with the highest concentration was BDE 47. Based on this information, the BDE flow was estimated for commerce, which is a form of pollutant transport not usually considered in POP pollution studies. A preliminary estimation indicated that the quantity of PBDEs mobilized by commerce was in the order of kg, and in the case of Chile might reach almost 1 kg.

Climate-induced changes to the ancestral population size of two Patagonian galaxiids: the influence of glacial cycling.

Patagonia is one of the few areas in the Southern Hemisphere to have been directly influenced by Quaternary glaciers. In this study, we evaluate the influence that Quaternary glacial ice had on the genetic diversity of two congeneric fish species, the diadromous Galaxias maculatus and the nondiadromous Galaxias platei, using multilocus estimates of effective population size through time. Mid-Quaternary glaciations had far-reaching consequences for both species. Galaxias maculatus and G. platei each experienced severe genetic bottlenecks during the period when Patagonia ice sheet advance reached its maximum positions c. 1.1-0.6 Ma. Concordant drops in effective size during this time suggest that range sizes were under similar constraints. It is therefore unlikely that coastal (brackish/marine) environments served as a significant refuge for G. maculatus during glacial periods. An earlier onset of population declines for G. platei suggests that this species was vulnerable to modest glacial advances. Declines in effective sizes were continuous for both species and lasted into the late-Pleistocene. However, G. maculatus exhibited a strong population recovery during the late-Quaternary (c. 400,000 bp). Unusually long and warm interglacials associated with the late-Quaternary may have helped to facilitate a strong population rebound in this primarily coastal species.

Ontogenetic shape trajectory of Trichomycterus areolatus varies in response to water velocity environment.

Body and head shape among fishes both vary between environments influenced by water velocity and across ontogeny. Although the shape changes associated with variation in average water velocity and ontogeny are well documented, few studies have tested for the interaction between these two variables (i.e., does ontogenetic shape variation differ between velocity environments). We use geometric morphometrics to characterize shape differences in Trichomycterus areolatus, a freshwater catfish found in high and low-velocity environments in Chile. We identify a significant interaction between velocity environment and body size (i.e., ontogeny). Ontogenetic patterns of shape change are consistent with other studies, but velocity environment differentially affects the ontogenetic trajectory of shape development in T. areolatus. Shape change over ontogeny appears more constrained in high-velocity environments compared to low-velocity environments.

Interplay of geomorphology and hydrology drives macroinvertebrate assemblage responses to hydropeaking.

Hydropeaking leads to major anthropogenic disturbance of river networks worldwide. Flow variations imposed by hydropeaking may significantly affect macroinvertebrate assemblages within the river network. As such, the responses of macroinvertebrate assemblages to hydropeaking are expected to be complex and vary across spatial and temporal scales as well as ecological organization levels. To unpack this complexity, we assessed the interplay of geomorphic and hydrological variables as drivers of the responses of macroinvertebrate assemblages to hydropeaking. Specifically, we studied different levels of ecological organization of macroinvertebrate assemblages in two functional process zones (FPZs; Sub-Andean and Central Valley Gravel Dominated) subjected to different flow management in two Chilean Andean river networks. Hydropeaking caused significant reduction of macroinvertebrate abundances in both FPZs and at all ecological organization levels with the exception of one feeding guild (scrapers). Furthermore, the response of macroinvertebrate assemblage variance was stronger in the Central Valley Gravel Dominated FPZ. Both geomorphic and hydrological variables influenced macroinvertebrate assemblage responses. However, the effects of the principal geomorphic variables operated at valley (meso) spatial scale and the main hydrological variables operated at the sub-daily (micro) temporal scale. Therefore, to minimise the effects of hydropeaking on macroinvertebrate assemblages, flow management should consider reduction of sub-daily variability. Furthermore, placement of new barriers should take into account not only their position within the river network but also their effects downstream that strongly depend on characteristics of river valley.

The longest fragment drives fish beta diversity in fragmented river networks: Implications for river management and conservation.

Connectivity plays a crucial role in maintaining the structural and functional attributes of river networks. Therefore, the loss of connectivity (fragmentation) alters the functioning and diversity patterns of the biota at local and regional scales. The global hydropower boom is one of the main drivers of river network fragmentation, with significant effects on the diversity of riverine biota. Analyses of beta diversity of fish assemblages in rivers with different degrees of fragmentation can give new insights into mechanisms that contribute to the responses of these assemblages to fragmentation. Here, fish beta diversity within six river networks of central Chile with different levels of fragmentation was studied to assess the responses of fish assemblages to fragmentation. A hypothesis of a significant effect of fragmentation on the beta diversity of native and non-native fish in riffles and pools was tested. This effect is expected to be modulated by both changes in environmental heterogeneity and direct obstruction of natural dispersal routes. Beta diversity based on variation of assemblage structure and environmental heterogeneity showed significant differences among river networks. Fish beta diversity showed a clear response to fragmentation in recently fragmented rivers. Specifically, the beta diversity of native fishes in pools and non-native fishes in riffles decreased with increase of the ratio between the longest non-fragmented sections of the river network to the total length of the network. These effects of fragmentation on fish assemblages were modulated by the biological features of each species, and open-water species were most severely affected. These results have significant implications for planning of the placement of new barriers in river networks subjected to hydropower boom. Planning of the placement of new barriers should consider the maintenance of long, connected sections within river networks in order to minimise the effects of fragmentation on fish biodiversity.

Surviving historical Patagonian landscapes and climate: molecular insights from Galaxias maculatus.

BACKGROUND: The dynamic geological and climatic histories of temperate South America have played important roles in shaping the contemporary distributions and genetic diversity of endemic freshwater species. We use mitochondria and nuclear sequence variation to investigate the consequences of mountain barriers and Quaternary glacial cycles for patterns of genetic diversity in the diadromous fish Galaxias maculatus in Patagonia (approximately 300 individuals from 36 locations). RESULTS: Contemporary populations of G. maculatus, east and west of the Andes in Patagonia, represent a single monophyletic lineage comprising several well supported groups. Mantel tests using control region data revealed a strong positive relationship when geographic distance was modeled according to a scenario of marine dispersal. (r = 0.69, P = 0.055). By contrast, direct distance between regions was poorly correlated with genetic distance (r = -0.05, P = 0.463). Hierarchical AMOVAs using mtDNA revealed that pooling samples according to historical (pre-LGM) oceanic drainage (Pacific vs. Atlantic) explained approximately four times more variance than pooling them into present-day drainage (15.6% vs. 3.7%). Further post-hoc AMOVA tests revealed additional genetic structure between populations east and west of the Chilean Coastal Cordillera (coastal vs. interior). Overall female effective population size appears to have remained relatively constant until roughly 0.5 Ma when population size rapidly increased several orders of magnitude [100x (60x-190x)] to reach contemporary levels. Maximum likelihood analysis of nuclear alleles revealed a poorly supported gene tree which was paraphyletic with respect to mitochondrial-defined haplogroups. CONCLUSIONS: First diversifying in the central/north-west region of Patagonia, G. maculatus extended its range into Argentina via the southern coastal regions that join the Pacific and Atlantic oceans. More recent gene flow between northern populations involved the most ancient and most derived lineages, and was likely facilitated by drainage reversal(s) during one or more cooling events of the late Pleistocene. Overall female effective population size represents the end result of a widespread and several hundred-fold increase over approximately 0.5 Ma, spanning several climatic fluctuations of the Pleistocene. The minor influence of glacial cycles on the genetic structure and diversity of G. maculatus likely reflects the access to marine refugia during repeated bouts of global cooling. Evidence of genetic structure that was detected on a finer scale between lakes/rivers is most likely the result of both biological attributes (i.e., resident non-migratory behavior and/or landlocking and natal homing in diadromous populations), and the Coastal Cordillera as a dispersal barrier.

Human-induced habitat fragmentation effects on connectivity, diversity, and population persistence of an endemic fish, Percilia irwini, in the Biobío River basin (Chile).

An understanding of how genetic variability is distributed in space is fundamental for the conservation and maintenance of diversity in spatially fragmented and vulnerable populations. While fragmentation can occur from natural barriers, it can also be exacerbated by anthropogenic activities such as hydroelectric power plant development. Whatever the source, fragmentation can have significant ecological effects, including disruptions of migratory processes and gene flow among populations. In Chile, the Biobío River basin exhibits a high degree of habitat fragmentation due to the numerous hydroelectric power plants in operation, the number of which is expected to increase following new renewable energy use strategies. Here, we assessed the effects of different kinds of barriers on the genetic structure of the endemic freshwater fish Percilia irwini, knowledge that is critically needed to inform conservation strategies in light of current and anticipated further fragmentation initiatives in the system. We identified eight genetic units throughout the entire Biobío system with high effective sizes. A reduced effective size estimate was, however, observed in a single population located between two impassable barriers. Both natural waterfalls and human-made dams were important drivers of population differentiation in this system; however, dams affect genetic diversity differentially depending on their mode of operation. Evidence of population extirpation was found in two river stretches limited by upstream and downstream dams. Significant gene flow in both directions was found among populations not separated by natural or anthropogenic barriers. Our results suggest a significant vulnerability of P. irwini populations to future dam development and demonstrate the importance of studying basin-wide data sets with genetic metrics to understand the strength and direction of anthropogenic impacts on fish populations.

Partitioning ß-diversity reveals that invasions and extinctions promote the biotic homogenization of Chilean freshwater fish fauna.

AIM: Exotic species' introductions together with extinction of native species represent the main mechanisms driving biotic homogenization of freshwater fish assemblages around the world. While generally ichtyofaunistic realms transit towards biotic homogenization, for conservation purposes it is essential to understand what specific mechanisms are promoting it on particular areas or regions. Here, we report the occurrence of biotic homogenization in 29 Chilean watersheds, analyzing its ß-diversity (including turnover and nestedness) and predicting future trends. LOCATION: Continental Chile (18o-56o S). METHODS: We determined fish composition per basin for historical and current assemblages; extant native, exotic, and extinct species were recorded as 1 (presence) or 0 (absence) in two matrices basins × species. For each matrix, we calculated the turnover (ßsim), nestedness (ßnes), and ß-diversity (ßsor); then, we obtained Δßsim, Δßnes, and Δßsor, as the arithmetical difference between basin pairs over time. In addition, we search for explanatory variables correlating Δßsim, Δßnes, and Δßsor with geographical and land use variables. Finally, simulating events of species introduction (i.e., invasion) and extinction, we generated 15 hypothetical assemblages, looking to establish future trends towards biotic change in Chilean basins. RESULTS: Species turnover and ß-diversity significantly decreased from historical to current assemblages (Δßsim = -0.084; Δßsor = -0.061, respectively), while the species nestedness did not show significant changes (Δßnes = 0.08). Biotic changes have been driven mainly by the introduction of 28 exotic species, with a minor role of extinctions (one species) and translocations (0 species) of native species. Changes in ß-diversity were negatively correlated with area, elevation, and geographical distance between basins but not with land-use nor human population. Finally, the analysis of 15 future assemblages predicts a significant decrease of ß-diversity and turnover, and an increase for species nestedness, this time promoted by an increase in the extinction of native species. MAIN CONCLUSION: Chilean basins show a significant decrease of the distributional ß-diversity and species turnover of the freshwater fish fauna, evidencing a trend towards biotic homogenization. This trend is shared with other Neotropical basins; however, specific mechanisms driving it show different magnitude. Changes in the ß-diversity components do not show correlation with variables associated to land use, thus suggesting that casual introductions of freshwater fishes in Chile follow an opportunistic mode related to commercial use. According to future scenarios simulated, biotic homogenization should increase further, mainly as consequence of increased native extinctions.

Biotransport of persistent organic pollutants in the southern Hemisphere by invasive Chinook salmon (Oncorhynchus tshawytscha) in the rivers of northern Chilean Patagonia, a UNESCO biosphere reserve.

Biotransport is often associated with migration patterns of species, including large, anadromous salmonids. Several studies have reported biotransport of persistent organic pollutants in the Northern Hemisphere, but there is no published information on biotransport ocurring south of the equator. Chile's Patagonia is one of the last largely intact natural areas in the world. The objective of this study was to determine whether persistent organic pollutants are transported by the invasive Pacific Chinook salmon (O. tshawytscha) from the Pacific Ocean to Chilean Patagonia. Samples of juvenile and adult Chinook salmon were analyzed for polychlorinated biphenyls, pesticides and polybrominated diphenyl ethers. The results revealed that concentrations of POPs in adults migrating into Patagonian rivers were significantly higher than those found in juveniles migrating seaward. A mass balance analysis indicates that Chinook salmon are a source of persistent organic pollutants to Chilean Patagonia inland waters. Capsule: Biotransport of Persistent Organic Pollutants (POPs) by Chinook salmon (O. tshawytscha) from the Pacific Ocean to Chilean Patagonia has been confirmed by mass balance of POPs.

Examining the influence of human stressors on benthic algae, macroinvertebrate, and fish assemblages in Mediterranean streams of Chile.

Changes in land-uses and cover are strong drivers of stream degradation worldwide. In Mediterranean ecoregions, land conversion from forest with native species to tree plantations, agricultural, and urban areas have drastically increased in recent years. Here, we examined the influence of land-use types on water quality (physical and chemical variables) and biota (benthic algae, macroinvertebrates, and fishes). We conducted field surveys between December-March 2016 (dry season) in 24 sites located across four land-use types including native vegetation (NV), tree plantation (TP), agriculture (Ag), and urban (Ur) within the Chilean Mediterranean ecoregion. We calculated metrics and multivariate statistics to assess correlations between biological and environmental variables. Significant differences between land-uses were found for environmental/physicochemical variables and the three biotic groups (i.e., benthic algae, macroinvertebrates, and fishes). We found higher chlorophyll-a biomass for TP, Ag, and Ur compared to NV streams. The macroinvertebrate diversity of sensitive taxa (i.e., Ephemeroptera, Plecoptera, and Trichoptera) and water quality were lower in land draining streams with human uses, other than NV, with the most evident impact found in Ag and Ur streams. NV and TP streams had a lower Diptera density and lower percentage of non-insect taxa. Fish richness and diversity were not affected by land-use. However, cold-water fishes (both native and introduced) were closely associated with NV and TP streams whereas warm-water fishes (mostly introduced) were closely associated with Ag and Ur streams. Multivariate analysis revealed that both local habitat and catchment-scale variables influenced each biological assemblage. Our findings suggest that benthic algae, macroinvertebrates, and fish populations are good indicators of human land-use disturbances, with Ag and Ur areas being the most detrimental for freshwater biota.