Diet of Andean leaf-eared mice (Phyllotis) living at extreme elevations on Atacama volcanoes: insights from metagenomics, DNA metabarcoding, and stable isotopes.

On the flanks of >6000 m Andean volcanoes that tower over the Atacama Desert, leaf-eared mice (Phyllotis vaccarum) live at extreme elevations that surpass known vegetation limits. What the mice eat in these barren, hyperarid environments has been the subject of much speculation. According to the arthropod fallout hypothesis, sustenance is provided by windblown insects that accumulate in snowdrifts ('aolian deposits'). It is also possible that mice feed on saxicolous lichen or forms of cryptic vegetation that have yet to be discovered at such high elevations. We tested hypotheses about the diet of mice living at extreme elevations on Atacama volcanoes by combining metagenomic and DNA metabarcoding analyses of gut contents with stable-isotope analyses of mouse tissues. Genomic analyses of contents of the gastrointestinal tract of a live-captured mouse from the 6739 m summit of Volcán Llullaillaco revealed evidence for an opportunistic but purely herbivorous diet, including lichens. Although we found no evidence of animal DNA in gut contents of the summit mouse, stable isotope data indicate that mice native to elevations at or near vegetation limits (~5100 m) include a larger fraction of animal prey in their diet than mice from lower elevations. Some plant species detected in the gut contents of the summit mouse are known to exist at lower elevations at the base of the volcano and in the surrounding Altiplano, suggesting that such plants may occur at higher elevations beneath the snowpack or in other cryptic microhabitats.

Non-indigenous species and their realized niche in tidepools along the South-East Pacific coast.

Non-indigenous species (NIS) have the potential to colonize and become established in a wide range of coastal habitats. Species with broad environmental tolerances can quickly adapt to local conditions and expand their niches along environmental gradients, and even colonize habitats with extreme abiotic conditions. Here we report and document the distribution of eight marine NIS (four seaweed and four invertebrate species) found in tidepools along a 3000 km latitudinal gradient along the Pacific coast of Chile (18.4°S to 41.9°S). The seaweed NIS Codium fragile, Capreolia implexa, Schottera nicaeensis and Mastocarpus latissimus were mostly distributed towards high latitudes (i.e., more southerly locations), where temperatures in tidepools were low. The invertebrate NIS Anemonia alicemartinae, Ciona robusta, Bugula neritina and Bugulina flabellata were more common towards low latitudes, where high temperatures were registered in the tidepools. Across the intertidal gradient, seaweed NIS were mostly found in pools in the mid and low intertidal zone, while invertebrate NIS occurred mostly in pools from the mid and upper intertidal zones. The realized niche spaces of NIS (based on the Outlying Mean Index, OMI) in the study area were mainly influenced by environmental conditions of temperature and salinity (along the latitudinal and intertidal gradients), while other tidepool characteristics (depth, surface area, exposition, and complexity) only had minor effects. Five of the eight NIS exhibited a realized niche space coinciding with the average tidepool environmental conditions, while marginal niches were occupied by species with affinities for specific temperatures and salinities along the latitudinal and intertidal gradients. Our results indicate that physiological tolerances to environmental factors play a fundamental role in the distribution of seaweed and invertebrate NIS in tidepools along the Chilean coast. This study confirms that tidepools offer suitable conditions for some seaweed and invertebrate NIS, potentially facilitating their invasion into new natural habitats.

Ecological variation in invasive brown trout (Salmo trutta) within a remote coastal river catchment in northern Patagonia complicates estimates of invasion impact.

Salmonids were first introduced into the Chilean fresh waters in the 1880s, and c. 140 years later, they are ubiquitous across Chilean rivers, especially in the southern pristine fresh waters. This study examined the brown trout (Salmo trutta) and native taxa ecology in two adjacent but contrasting rivers of Chilean Patagonia. During spring 2016 and spring-fall 2017 we examined the variation in benthic macroinvertebrate and fish community composition and characterized fish size structure, stomach contents, and stable isotopes (δ13 C and δ15 N) to understand population structure, fish diet, and trophic interactions between S. trutta and native taxa. The native Galaxias maculatus (puye) dominated the fish community (74% of abundance). S. trutta was less abundant (16% of survey catch) but dominated the fish community (over 53%) in terms of biomass. S. trutta showed distinct diets (stomach content analysis) in the two rivers, and individuals from the larger river were notably more piscivorous, consuming native fish with a relatively small body size (<100-mm total length). Native fishes were isotopically distinct from S. trutta, which showed a wider isotopic niche in the smaller river, indicating that their trophic role was more variable than in the larger river (piscivorous). This study provides data from the unstudied pristine coastal rivers in Patagonia and reveals that interactions between native and introduced species can vary at very local spatial scales.

Multiple stable isotopes (C, N & S) provide evidence for fin whale (Balaenoptera physalus) trophic ecology and movements in the Humboldt Current System of northern Chile.

Reflecting the intense coastal upwelling and high primary productivity characteristic of the Humboldt Current System (HCS), the northern coast of Chile supports a diverse and productive community of marine consumers, including worldwide important pelagic fisheries resources. Although marine mammals are relatively understudied in the region, recent studies have demonstrated that fin whale (Balaenoptera physalus) is the most frequently encountered whale species, and forages in these waters year-round. However, a current lack of information limits our understanding of whether fin whales actively feed and/or remain resident in these waters or whether whales are observed feeding as they migrate along this part of the Pacific. Here, we use stable isotope ratios of carbon, nitrogen and sulphur of fin whale skin samples collected in early summer 2020 (n = 18) and in late winter 2021 (n = 22) to examine evidence of temporal isotopic shifts that could provide information on potential migratory movements and to estimate likely consumption patterns of putative prey (i.e. zooplankton, krill, pelagic fishes and Pleuroncodes sp.). We also analysed prey items in fin whale faecal plumes (n = 8) collected during the study period. Stable isotope data showed significant differences in the isotopic values of fin whales from summer and winter. On average, summer individuals were depleted in 15N and 34S relative to those sampled during winter. Whales sampled in summer showed greater isotopic variance than winter individuals, with several showing values that were atypical for consumers from the HCS. During winter, fin whales showed far less inter-individual variation in stable isotope values, and all individuals had values indicative of prey consumption in the region. Analysis of both stable isotopes and faeces indicated that fin whales sighted off the Mejillones Peninsula fed primarily on krill (SIA median contribution = 32%; IRI = 65%) and, to a lesser extent, zooplankton (SIA zooplankton = 29%; IRI copepod = 33%). These are the first isotopic-based data regarding the trophic ecology of fin whales in the north of Chile. They provide evidence that fin whales are seasonally resident in the area, including individuals with values that likely originated outside the study area. The information presented here serves as a baseline for future work. It highlights that many aspects of the ecology of fin whales in the Humboldt Current and wider SE Pacific still need to be clarified.

No evidence for effects of mill-grinding on δ13 C, δ15 N and δ34 S values in different marine taxa.

RATIONALE: The analysis of natural variation in light stable isotopes such as carbon (δ13 C), nitrogen (δ15 N) and sulfur (δ34 S) plays an important role in deepening our understanding of ecosystems. To avoid misinterpretation, robust results are required, where pre-treatment steps such as sample homogenization are crucial to guarantee representative samples. The grinding of samples using stainless steel balls in polypropylene tubes (e.g., laboratory bead-beater) has been identified as a potential source of bias. METHODS: We tested possible effects of mill-grinding (e.g., contamination) of samples of coastal marine taxa including primary producers, primary consumers and higher trophic level fish. We compared potential impacts of homogenization by mill-grinding with hand-grinding over an extended time on δ13 C, δ15 N and δ34 S values. RESULTS: One-way Welch's analysis of variance (ANOVA) showed that there were no statistical differences between methods for all the studied taxa. Also, repeated measures ANOVA showed no evidence of effects of grinding for extended times (from 30 to 120 seconds) for δ13 C, δ15 N and δ34 S values. CONCLUSIONS: We found no evidence that grinding samples in polypropylene tubes in a bead-beater resulted in any marked alteration of the isotopic composition on the studied samples, e.g., through contamination by plastic. As such, we consider mill-grinding as an appropriate method for the homogenization of samples from a range of different marine taxa, which under controlled conditions did not affect δ13 C, δ15 N and δ34 S analysis.

'White gold' guano fertilizer drove agricultural intensification in the Atacama Desert from AD 1000.

The archaeological record shows that large pre-Inca agricultural systems supported settlements for centuries around the ravines and oases of northern Chile's hyperarid Atacama Desert. This raises questions about how such productivity was achieved and sustained, and its social implications. Using isotopic data of well-preserved ancient plant remains from Atacama sites, we show a dramatic increase in crop nitrogen isotope values (δ15N) from around AD 1000. Maize was most affected, with δ15N values as high as +30‰, and human bone collagen following a similar trend; moreover, their carbon isotope values (δ13C) indicate a considerable increase in the consumption of maize at the same time. We attribute the shift to extremely high δ15N values-the highest in the world for archaeological plants-to the use of seabird guano to fertilize crops. Guano-'white gold' as it came to be called-thus sustained agricultural intensification, supporting a substantial population in an otherwise extreme environment.

Functional changes in benthic macrofaunal communities along a natural gradient of hypoxia in an upwelling system.

Increasing global concern has been raised about the expansion of hypoxia in coastal waters and its potential to impact benthic ecosystems. Upwelling areas offer opportunities to study the effects of hypoxia on benthic communities under natural conditions. We used a biological trait-based approach and estimated functional diversity indices to assess macrobenthic community functioning along a depth gradient associated with naturally increasing hypoxia and concentrations of organic matter in the upwelling zone of northern Chile (South-East Pacific) over two years. Our results highlighted the increasing dominance of opportunistic biological traits associated with hypoxia and high organic matter content. Habitat filtering was the main process affecting the studied communities. Functional diversity patterns were persistent overtime despite the occurrence of a pulse of oxygenation. This study contributes to our understanding of how natural hypoxia impacts macrobenthic communities, providing useful information in the context of increasing eutrophication due to human influence on coastal areas.

Intraspecific variation and energy channel coupling within a Chilean kelp forest.

The widespread importance of variable types of primary production, or energy channels, to consumer communities has become increasingly apparent. However, the mechanisms underlying this "multichannel" feeding remain poorly understood, especially for aquatic ecosystems that pose unique logistical constraints given the diversity of potential energy channels. Here, we use bulk tissue isotopic analysis along with carbon isotope (δ13 C) analysis of individual amino acids to characterize the relative contribution of pelagic and benthic energy sources to a kelp forest consumer community in northern Chile. We measured bulk tissue δ13 C and δ15 N for >120 samples; of these we analyzed δ13 C values of six essential amino acids (EAA) from nine primary producer groups (n = 41) and 11 representative nearshore consumer taxa (n = 56). Using EAA δ13 C data, we employed linear discriminant analysis (LDA) to assess how distinct EAA δ13 C values were between local pelagic (phytoplankton/particulate organic matter), and benthic (kelps, red algae, and green algae) endmembers. With this model, we were able to correctly classify nearly 90% of producer samples to their original groupings, a significant improvement on traditional bulk isotopic analysis. With this EAA isotopic library, we then generated probability distributions for the most important sources of production for each individual consumer and species using a bootstrap-resampling LDA approach. We found evidence for multichannel feeding within the community at the species level. Invertebrates tended to focus on either pelagic or benthic energy, deriving 13-67% of their EAA from pelagic sources. In contrast, mobile (fish) taxa at higher trophic levels used more equal proportions of each channel, ranging from 19% to 47% pelagically derived energy. Within a taxon, multichannel feeding was a result of specialization among individuals in energy channel usage, with 37 of 56 individual consumers estimated to derive >80% of their EAA from a single channel. Our study reveals how a cutting-edge isotopic technique can characterize the dynamics of energy flow in coastal food webs, a topic that has historically been difficult to address. More broadly, our work provides a mechanism as to how multichannel feeding may occur in nearshore communities, and we suggest this pattern be investigated in additional ecosystems.

Chilean Salmon Sushi: Genetics Reveals Product Mislabeling and a Lack of Reliable Information at the Point of Sale.

Species diagnosis is essential to assess the level of mislabeling or misnamed seafood products such as sushi. In Chile, sushi typically includes salmon as the main ingredient, but species used are rarely declared on the menu. In order to identify which species are included in the Chilean sushi market, we analyzed 84 individual sushi rolls sold as "salmon" from sushi outlets in ten cities across Chile. Using a polymerase chain reaction-restriction fragment length polymorphism protocol (PCR-RFLP), we identified mislabeled and misnamed products. Atlantic salmon was the most common salmonid fish used in sushi, followed by coho salmon, rainbow trout, and Chinook salmon. We found a total of 23% and 18% of the products were mislabeled and misnamed, respectively. In 64% of cases, the salesperson selling the product could not identify the species. We also identified the use of wild-captured Chinook salmon samples from a naturalized population. Our results provide a first indication regarding species composition in Chilean sushi, a quantification of mislabeling and the level of misinformation declared by sales people to consumers. Finally, considering that Chinook salmon likely originates from a non-licensed origin and that sushi is an uncooked product, proper identification in the food production chain may have important consequences for the health of consumers.

Southernmost distribution limit for endangered Peladillas (Aplochiton taeniatus) and non-native coho salmon (Oncorhynchus kisutch) coexisting within the Cape Horn biosphere reserve, Chile.

The Cape Horn Biosphere Reserve, one of the last wild areas of the planet, is not exempt from the pressures of global change, such as non-native species introductions. During 2018 and 2019 we studied the Róbalo river basin in order to update the diversity and distribution of fishes. Here, we report for the first time the native and endangered "Peladillas" Aplochiton taeniatus and the non-native coho salmon Oncorhynchus kisutch. The coexistence of native and non-native fishes poses a challenge for the management and conservation of aquatic biota from the Cape Horn Biosphere Reserve.

Sample acidification has a predictable effect on isotopic ratios of particulate organic matter along the Chilean coast.

RATIONALE: Stable isotopes of carbon and nitrogen have proved to be valuable tools for researchers working across the different subfields of ecology. However, the chemical pretreatment of samples prior to analytical determination of stable isotope ratios can influence the results, and therefore conclusions regarding the ecology of the taxon or system under study. Here, we determined the effect of vapor acidification with concentrated HCl on the δ13 C and δ15 N values of particulate organic matter (POM), which are commonly used as baselines for studies of trophic ecology, or to understand oceanographic patterns. METHODS: Samples of marine POM were obtained along a large-scale latitudinal gradient (ca 3000 km) along the Chilean coast, along with a range of oceanographic variables thought to potentially influence inorganic carbon at each sampling location. A random subset of 50 samples was divided into two parts: one acidified by HCl fumigation treatment, and the other acting as a control. We compared paired differences in δ13 C and δ15 N values measured by continuous flow isotope ratio mass spectrometry and used a model selection approach to examine which oceanographic factor best explained shifts in values following acid treatment. RESULTS: Acidification resulted in statistically significant reductions in both δ13 C and δ15 N values, but the effect was relatively small. The model that best explained the differences between acidified and non-acidified δ13 C values included depth, salinity and sea-surface temperature at the sampling point. A regression of acidified on non-acidified δ13 C values shows that the treatment effect was strongest on samples more depleted in 13 C. CONCLUSIONS: The differences between δ13 C and δ15 N values in acidified and non-acidified samples are linear and predictable. This implies that the nature of the POM and its possible alteration during the acid treatment are important factors that support the reliable determination of the values of δ13 C and δ15 N of POM.

Clarifying a trophic black box: stable isotope analysis reveals unexpected dietary variation in the Peruvian anchovy Engraulis ringens.

BACKGROUND: Small fishes play fundamental roles in pelagic ecosystems, channelling energy and nutrients from primary producers to higher trophic levels. They support globally important fisheries in eastern boundary current ecosystems like the Humboldt Current System (HCS) of the SE Pacific (Chile and Peru), where fish catches are the highest in the world (per unit area). This production is associated with coastal upwelling where fisheries target small pelagic fishes including the Peruvian anchovy (Engraulis ringens). The elevated biomass attained by small pelagics is thought to reflect their low trophic position in short/simple food chains. Despite their global importance, large gaps exist in our understanding of the basic ecology of these resources. For instance, there is an ongoing debate regarding the relative importance of phytoplankton versus animal prey in anchovy diet, and ecosystem models typically assign them a trophic position (TP) of ∼2, assuming they largely consume phytoplankton. Recent work based on both relative energetic content and stable isotope analysis (SIA) suggests that this value is too low, with δ 15N values indicating that anchovy TP is ca. 3.5 in the Peruvian HCS. METHODS: We characterised the trophic ecology of adult anchovies (n = 30), their putative prey and carnivorous jack mackerel (n = 20) captured from N Chile. SIA (δ 13C and δ 15N) was used to estimate the relative contribution of different putative prey resources. δ 15N was used to estimate population level trophic position. RESULTS: Anchovies showed little variability in δ 13C (-18.7 to -16.1‰) but varied greatly in δ 15N (13.8 to 22.8‰)-individuals formed two groups with low or high δ 15N values. When considered as a single group, mixing models indicated that anchovy diet was largely composed of zooplankton (median contribution: 95% credibility limits), with major contributions of crustacean larvae (0.61: 0.37-0.77) and anchovy (preflexion) larvae (0.15: 0.02-0.34), and the assimilation of phytoplankton was negligible (0.05: 0-0.22). The modal (95% credibility limits) estimate of TP for the pooled anchovy sample was 3.23 (2.93-3.58), overlapping with recent SIA-based estimates from Peru. When the two δ 15N groups were analysed separately, our results indicate that the lower δ 15N group largely assimilated materials from crustacean larvae (0.73: 0.42-0.88), with a TP of 2.91 (2.62-3.23). Mixing models suggested high δ 15N anchovies were cannibalistic, consuming anchovy preflexion larvae (0.55: 0.11-0.74). A carnivorous trophic niche was supported by high TP (3.79: 3.48-4.16), mirroring that of carnivorous juvenile jack mackerel (Trachurus murphyi; 3.80: 3.51-4.14). Our results support recent conclusions regarding high TP values of anchovy from Peru and reveal new insights into their trophic behaviour. These results also highlight the existence of cryptic trophic complexity and ecosystem function in pelagic food webs, classically considered as simple.

Geologic and anthropogenic sources of contamination in settled dust of a historic mining port city in northern Chile: health risk implications.

Chile is the leading producer of copper worldwide and its richest mineral deposits are found in the Antofagasta Region of northern Chile. Mining activities have significantly increased income and employment in the region; however, there has been little assessment of the resulting environmental impacts to residents. The port of Antofagasta, located 1,430 km north of Santiago, the capital of Chile, functioned as mineral stockpile until 1998 and has served as a copper concentrate stockpile since 2014. Samples were collected in 2014 and 2016 that show elevated concentrations of As, Cu, Pb, and Zn in street dust and in residents' blood (Pb) and urine (As) samples. To interpret and analyze the spatial variability and likely sources of contamination, existent data of basement rocks and soil geochemistry in the city as well as public-domain airborne dust were studied. Additionally, a bioaccessibility assay of airborne dust was conducted and the chemical daily intake and hazard index were calculated to provide a preliminary health risk assessment in the vicinity of the port. The main conclusions indicate that the concentrations of Ba, Co, Cr, Mn, Ni, and V recorded from Antofagasta dust likely originate from intrusive, volcanic, metamorphic rocks, dikes, or soil within the city. However, the elevated concentrations of As, Cd, Cu, Mo, Pb, and Zn do not originate from these geologic outcrops, and are thus considered anthropogenic contaminants. The average concentrations of As, Cu, and Zn are possibly the highest in recorded street dust worldwide at 239, 10,821, and 11,869 mg kg-1, respectively. Furthermore, the contaminants As, Pb, and Cu exhibit the highest bioaccessibilities and preliminary health risk indices show that As and Cu contribute to elevated health risks in exposed children and adults chronically exposed to dust in Antofagasta, whereas Pb is considered harmful at any concentration. Therefore, an increased environmental awareness and greater protective measures are necessary in Antofagasta and possibly other similar mining port cities in developing countries.

Microbial community composition and trophic role along a marked salinity gradient in Laguna Puilar, Salar de Atacama, Chile.

The geological, hydrological and microbiological features of the Salar de Atacama, the most extensive evaporitic sedimentary basin in the Atacama Desert of northern Chile, have been extensively studied. In contrast, relatively little attention has been paid to the composition and roles of microbial communities in hypersaline lakes which are a unique feature in the Salar. In the present study biochemical, chemical and molecular biological tools were used to determine the composition and roles of microbial communities in water, microbial mats and sediments along a marked salinity gradient in Laguna Puilar which is located in the "Los Flamencos" National Reserve. The bacterial communities at the sampling sites were dominated by members of the phyla Bacteroidetes, Chloroflexi, Cyanobacteria and Proteobacteria. Stable isotope and fatty acid analyses revealed marked variability in the composition of microbial mats at different sampling sites both horizontally (at different sites) and vertically (in the different layers). The Laguna Puilar was shown to be a microbially dominated ecosystem in which more than 60% of the fatty acids at particular sites are of bacterial origin. Our pioneering studies also suggest that the energy budgets of avian consumers (three flamingo species) and dominant invertebrates (amphipods and gastropods) use minerals as a source of energy and nutrients. Overall, the results of this study support the view that the Salar de Atacama is a heterogeneous and fragile ecosystem where small changes in environmental conditions may alter the balance of microbial communities with possible consequences at different trophic levels.

The activity of nitrifying microorganisms in a high-altitude Andean wetland.

High-altitude wetland holds freshwater springs, evaporitic ponds and lagoon with variable salinity and nutrients, potentially influencing the ecology of nitrifying communities. In this study, nitrifying microorganisms in Salar de Huasco (Chile) were surveyed to determine bacterial and archaeal contribution to ammonium (AO), nitrite oxidation (NO), ammonium uptake (AU) during wet and dry seasons. The activity signals from these groups were assessed by specific amoA-qPCR transcription, 15N tracer studies and addition of group specific inhibitor experiments for nitrifying microorganisms (N1-guanyl-1, 7-diaminoheptane [GC7]-archaeal specific and allylthiourea [ATU]-bacterial specific). Nitrifying communities, i.e. Nitrosopumilus, Nitrosospira, Nitrosomonas, Kuenenia and Nitrospira, were more frequent (∼0.25% of 16S rRNA sequences) at low salinity sites. Bacterial amoA-qPCR transcripts also increased at low salinity and along in situ ammonium increase observed between wet/dry seasons. Nutrient changes through time and 15N tracer experiments results showed that AO and NO were detected and peaked mainly at low salinity-high ammonium sites (<37 000 µS cm-1 and >0.3 µM), whereas AU was predominant at evaporitic sites. Our results indicate that salinity and ammonium affect the nitrifying communities that are potentially more active at low-salinity sites but persistent at saltier evaporitic areas of the wetland when ammonium is available.

Habitat coupling writ large: pelagic-derived materials fuel benthivorous macroalgal reef fishes in an upwelling zone.

Coastal marine upwelling famously supports elevated levels of pelagic biological production, but can also subsidize production in inshore habitats via pelagic-benthic coupling. Consumers inhabiting macroalgae-dominated rocky reef habitats are often considered to be members of a food web fuelled by energy derived from benthic primary production; conversely, they may also be subsidized by materials transported from pelagic habitats. Here, we used stable isotopes (δ13 C, δ15 N) to examine the relative contribution of pelagic and benthic materials to an ecologically and economically important benthivorous fish assemblage inhabiting subtidal macroalgae-dominated reefs along ~1,000 km of the northern Chilean coast where coastal upwelling is active. Fish were isotopically most similar to the pelagic pathway and Bayesian mixing models indicated that production of benthivorous fish was dominated (median 98%, range 69-99%) by pelagic-derived C and N. Although the mechanism by which these materials enter the benthic food web remains unknown, our results clearly highlight the importance of pelagic-benthic coupling in the region. The scale of this subsidy has substantial implications for our basic understanding of ecosystem functioning and the management of nearshore habitats in northern Chile and other upwelling zones worldwide.

Bacterial Active Community Cycling in Response to Solar Radiation and Their Influence on Nutrient Changes in a High-Altitude Wetland.

Microbial communities inhabiting high-altitude spring ecosystems are subjected to extreme changes in solar irradiance and temperature throughout the diel cycle. Here, using 16S rRNA gene tag pyrosequencing (cDNA) we determined the composition of actively transcribing bacteria from spring waters experimentally exposed through the day (morning, noon, and afternoon) to variable levels of solar radiation and light quality, and evaluated their influence on nutrient recycling. Solar irradiance, temperature, and changes in nutrient dynamics were associated with changes in the active bacterial community structure, predominantly by Cyanobacteria, Verrucomicrobia, Proteobacteria, and 35 other Phyla, including the recently described Candidate Phyla Radiation (e.g., Parcubacteria, Gracilibacteria, OP3, TM6, SR1). Diversity increased at noon, when the highest irradiances were measured (3.3-3.9 H', 1125 W m-2) compared to morning and afternoon (0.6-2.8 H'). This shift was associated with a decrease in the contribution to pyrolibraries by Cyanobacteria and an increase of Proteobacteria and other initially low frequently and rare bacteria phyla (< 0.5%) in the pyrolibraries. A potential increase in the activity of Cyanobacteria and other phototrophic groups, e.g., Rhodobacterales, was observed and associated with UVR, suggesting the presence of photo-activated repair mechanisms to resist high levels of solar radiation. In addition, the percentage contribution of cyanobacterial sequences in the afternoon was similar to those recorded in the morning. The shifts in the contribution by Cyanobacteria also influenced the rate of change in nitrate, nitrite, and phosphate, highlighted by a high level of nitrate accumulation during hours of high radiation and temperature associated with nitrifying bacteria activity. We did not detect ammonia or nitrite oxidizing bacteria in situ, but both functional groups (Nitrosomona and Nitrospira) appeared mainly in pyrolibraries generated from dark incubations. In total, our results reveal that both the structure and the diversity of the active bacteria community was extremely dynamic through the day, and showed marked shifts in composition that influenced nutrient recycling, highlighting how abiotic variation affects potential ecosystem functioning.

Coastal Upwelling Drives Intertidal Assemblage Structure and Trophic Ecology.

Similar environmental driving forces can produce similarity among geographically distant ecosystems. Coastal oceanic upwelling, for example, has been associated with elevated biomass and abundance patterns of certain functional groups, e.g., corticated macroalgae. In the upwelling system of Northern Chile, we examined measures of intertidal macrobenthic composition, structure and trophic ecology across eighteen shores varying in their proximity to two coastal upwelling centres, in a hierarchical sampling design (spatial scales of >1 and >10 km). The influence of coastal upwelling on intertidal communities was confirmed by the stable isotope values (δ13C and δ15N) of consumers, including a dominant suspension feeder, grazers, and their putative resources of POM, epilithic biofilm, and macroalgae. We highlight the utility of muscle δ15N from the suspension feeding mussel, Perumytilus purpuratus, as a proxy for upwelling, supported by satellite data and previous studies. Where possible, we used corrections for broader-scale trends, spatial autocorrelation, ontogenetic dietary shifts and spatial baseline isotopic variation prior to analysis. Our results showed macroalgal assemblage composition, and benthic consumer assemblage structure, varied significantly with the intertidal influence of coastal upwelling, especially contrasting bays and coastal headlands. Coastal topography also separated differences in consumer resource use. This suggested that coastal upwelling, itself driven by coastline topography, influences intertidal communities by advecting nearshore phytoplankton populations offshore and cooling coastal water temperatures. We recommend the isotopic values of benthic organisms, specifically long-lived suspension feeders, as in situ alternatives to offshore measurements of upwelling influence.

Parsing parallel evolution: ecological divergence and differential gene expression in the adaptive radiations of thick-lipped Midas cichlid fishes from Nicaragua.

The study of parallel evolution facilitates the discovery of common rules of diversification. Here, we examine the repeated evolution of thick lips in Midas cichlid fishes (the Amphilophus citrinellus species complex)-from two Great Lakes and two crater lakes in Nicaragua-to assess whether similar changes in ecology, phenotypic trophic traits and gene expression accompany parallel trait evolution. Using next-generation sequencing technology, we characterize transcriptome-wide differential gene expression in the lips of wild-caught sympatric thick- and thin-lipped cichlids from all four instances of repeated thick-lip evolution. Six genes (apolipoprotein D, myelin-associated glycoprotein precursor, four-and-a-half LIM domain protein 2, calpain-9, GTPase IMAP family member 8-like and one hypothetical protein) are significantly underexpressed in the thick-lipped morph across all four lakes. However, other aspects of lips' gene expression in sympatric morphs differ in a lake-specific pattern, including the magnitude of differentially expressed genes (97-510). Generally, fewer genes are differentially expressed among morphs in the younger crater lakes than in those from the older Great Lakes. Body shape, lower pharyngeal jaw size and shape, and stable isotopes (δ(13)C and δ(15)N) differ between all sympatric morphs, with the greatest differentiation in the Great Lake Nicaragua. Some ecological traits evolve in parallel (those related to foraging ecology; e.g. lip size, body and head shape) but others, somewhat surprisingly, do not (those related to diet and food processing; e.g. jaw size and shape, stable isotopes). Taken together, this case of parallelism among thick- and thin-lipped cichlids shows a mosaic pattern of parallel and nonparallel evolution.