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Summary: Phenotypic and compositional changes of immune cells in cerebrospinal fluid (CSF) can be used as biomarkers to help diagnose and track disease activity for neuroinflammatory and neurodegenerative diseases. Here, we present a workflow to perform high-dimensional immune profiling at single-cell resolution using cytometry by time-of-flight (CyTOF) on cells isolated from the CSF of patients with neuroinflammation. We describe steps for sample collection and preparation, barcoding to allow for multiplexing, and downstream data analysis using R.For complete details on the use and execution of this protocol, please refer to Fernández-Zapata et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Abstract Disease-modifying therapies (DMTs) are widely used in neuroimmunological diseases such as multiple sclerosis (MS) and neuromyelitis optica spectrum disorder (NMOSD). Although these treatments are known to predispose patients to infections and affect their responses to vaccination, little is known about the impact of DMTs on the myeloid cell compartment. In this study, we use mass cytometry to examine DMT-associated changes in the innate immune system in untreated and treated patients with MS (n = 39) or NMOSD (n = 23). We also investigated the association between changes in myeloid cell phenotypes and longitudinal responsiveness to homologous primary, secondary, and tertiary SARS-CoV-2 mRNA vaccinations. Multiple DMT-associated myeloid cell clusters, in particular CD64+HLADRlow granulocytes, showed significant correlations with B and T cell responses induced by vaccination. Our findings suggest the potential role of myeloid cells in cellular and humoral responses following vaccination in DMT-treated patients with neuroimmunological diseases.

Multiple state-of-the-art analyses of immune cells in 117 blood, 117 cerebrospinal fluid, 13 choroid plexus and 13 brain parenchyma samples reveal differential characteristics of immune cells in different body compartments and different diseases.

Abstract Inflammatory bowel disease (IBD) is characterized by dysregulated intestinal immune responses. Using mass cytometry (CyTOF) to analyze the immune cell composition in the lamina propria (LP) of patients with ulcerative colitis (UC) and Crohn's disease (CD), we observed an enrichment of CD4+ effector T cells producing IL‐17A and TNF, CD8+ T cells producing IFNγ, T regulatory (Treg) cells, and innate lymphoid cells (ILC). The function of these immune cells is regulated by store‐operated Ca2+ entry (SOCE), which results from the opening of Ca2+ release‐activated Ca2+ (CRAC) channels formed by ORAI and STIM proteins. We observed that the pharmacologic inhibition of SOCE attenuated the production of proinflammatory cytokines including IL‐2, IL‐4, IL‐6, IL‐17A, TNF, and IFNγ by human colonic T cells and ILCs, reduced the production of IL‐6 by B cells and the production of IFNγ by myeloid cells, but had no effect on the viability, differentiation, and function of intestinal epithelial cells. T cell‐specific deletion of CRAC channel genes in mice showed that Orai1, Stim1, and Stim2‐deficient T cells have quantitatively distinct defects in SOCE, which correlate with gradually more pronounced impairment of cytokine production by Th1 and Th17 cells and the severity of IBD. Moreover, the pharmacologic inhibition of SOCE with a selective CRAC channel inhibitor attenuated IBD severity and colitogenic T cell function in mice. Our data indicate that SOCE inhibition may be a suitable new approach for the treatment of IBD.

The activity of marine microorganisms depends on community composition, yet, in some oceans, less is known about the environmental and ecological processes that structure their distribution. The objective of this study was to test the effect of geographical distance and environmental parameters on prokaryotic community structure in the Southern Ocean (SO). We described the total (16S rRNA gene) and the active fraction (16S rRNA-based) of surface microbial communities over a ~6,500 km longitudinal transect in the SO. We found that the community composition of the total fraction was different from the active fraction across the zones investigated. In addition, higher α-diversity and stronger species turnover were displayed in the active community compared to the total community. Oceanospirillales, Alteromonadales, Rhodobacterales, and Flavobacteriales dominated the composition of the bacterioplankton communities; however, there were marked differences at the order level. Temperature, salinity, silicic acid, particulate organic nitrogen, and particulate organic carbon correlated with the composition of bacterioplankton communities. A strong distance–decay pattern between closer and distant communities was observed. We hypothesize that it was related to the different oceanic fronts present in the Antarctic Circumpolar Current. Our findings contribute to a better understanding of the complex arrangement that shapes the structure of bacterioplankton communities in the SO.

La puesta en valor del universo de los lenguajes estéticos con relación a las necesidades de la sociedad contemporánea y el permanente cuestionamiento de la vigencia de las estructuras educativas tradicionales desemboca en la búsqueda de nuevas alternativas pedagógicas. En este sentido, se reflexiona sobre la necesidad de entretejer los ámbitos artísticos, simbólicos y estéticos con las diversas áreas de aprendizaje, a la luz de perspectivas que tengan la intención de resignificar las dimensiones que configuran los procesos educativos. Además, se vislumbra la importancia de generar oportunidades formativas que atiendan a la figura docente con la intención de que se establezca como el vector fundamental de transformación y mejora de la calidad educativa, a partir de las experiencias cognitivas y emocionales que se generan desde la expresión estética.

Abstract Myeloid cells contribute to inflammation and demyelination in the early stages of multiple sclerosis (MS), but it is still unclear to what extent these cells are involved in active lesion formation in progressive MS (PMS). Here, we have harnessed the power of single-cell mass cytometry (CyTOF) to compare myeloid cell phenotypes in active lesions of PMS donors with those in normal-appearing white matter from the same donors and control white matter from non-MS donors. CyTOF measurements of a total of 74 targeted proteins revealed a decreased abundance of homeostatic and TNFhi microglia, and an increase in highly phagocytic and activated microglia states in active lesions of PMS donors. Interestingly, in contrast to results obtained from studies of the inflammatory early disease stages of MS, infiltrating monocyte-derived macrophages were scarce in active lesions of PMS, suggesting fundamental differences of myeloid cell composition in advanced stages of PMS.

Microorganisms play a crucial role in biogeochemical processes affecting the primary production and biogeochemical cycles of the ocean. In subpolar areas, the increment of the water temperature induced by climate change could lead to changes in the structure and activity of planktonic microbial communities. To understand how the structure of the microbial community in Chilean Patagonian fjords could be affected by climate change, we analyzed the composition of the prokaryotic community (bacteria-archaea) in two fjords (Pia and Yendegaia) with contrasting morphological and hydrological features. We targeted both the standing stock (16S rRNA genes) and the active fraction (16S rRNA transcripts) of the microbial communities during two consecutive austral winters. Our results showed that in both fjords, the active community had higher diversity and stronger biogeographic patterns when compared to the standing stock. Members of the Alpha-, Gamma-, and Deltaproteobacteria followed by archaea from the Marine Group I (Thaumarchaeota) dominated the active communities in both fjords. However, in Pia fjord, which has a marine-terminating glacier, the composition of the microbial community was directly influenced by the freshwater discharges from the adjacent glacier, and indirectly by a possible upwelling phenomenon that could bring deep sea bacteria such as SAR202 to the surface layer. In turn, in the Yendegaia, which has a land-terminating glacier, microbial communities were more similar to the ones described in oceanic waters. Furthermore, in Yendegaia fjord, inter-annual differences in the taxonomic composition and diversity of the microbial community were observed. In conclusion, Yendegaia fjord, without glacier calving, represents a fjord type that will likely be more common under future climate scenarios. Our results showing distinct Yendegaia communities, with for example more potential nitrogen-fixing microorganisms (Planctomycetes), indicate that as a result of climate change, changing planktonic communities could potentially impact biogeochemical processes and nutrient sources in subantarctic fjords.

En el marco de la reflexión sobre los distintos lenguajes de la literatura infantil, se percibe la necesidad de una aproximación a los aspectos semióticos en la mediación de la lectura. Para ello, se ofrece una revisión teórica que abarca la configuración del libro ilustrado como objeto significativo, la articulación de los códigos visuales con el texto escrito en la construcción de sentidos y la implicación de estos lenguajes en la comprensión de los contenidos literarios. Además, se aborda la relevancia de la figura mediadora como puente entre el libro y la comunidad, y se analizan diversos recursos insertos en sus propuestas estéticas en interrelación con el público y el contexto. En el ámbito metodológico, bajo la mirada cualitativa de investigación, se utiliza la técnica de grupo de discusión que, en este caso, recoge los aportes de exponentes latinoamericanos del campo de la mediación de la lectura. Tras revisar los postulados teóricos a la luz de las reflexiones suscitadas por el estudio, se establece que en el proceso de mediación se añaden nuevos elementos de sentido y significación, al complementar los contenidos del libro con otros lenguajes que subyacen en cada apropiación, interpretación y conformación del acto de lectura.

Chilean native forests have been subjected to several types of disturbances, with one of them being the replacement by exotic species. Pinus radiata D. Don is a widespread exotic tree that forms extensive plantations in southern Chile. It covers extended areas, affecting the landscape, biodiversity, and ecosystem services associated with native forest ecosystems. Although advances in assessing the impact of exotic plant species have been conducted, few studies have focused on the alteration of soil microorganisms. This study aimed to characterize the rhizosphere bacterial and fungal communities associated with the tree species Nothofagus obliqua inside a native forest stand and within a P. radiata plantation growing nearby. We used a 16S rRNA gene and ITS region metabarcoding approach. Using bioinformatics, diversity indices, relative abundance, preferential taxa, and predicted functions and guilds were estimated. The β-diversity analysis showed that both factors, the type of soil (rhizosphere or bulk soil) and the type of site (native forest or P. radiata plantation), were significant, with the site explaining most of the variation among bacterial and fungal communities. Proteobacteria and Basidiomycota were the most abundant bacterial and fungal phyla in both types of soil and sites. Similarly, bacteria showed similar abundant taxa at the family level, independent of the soil type or the site. The main fungal taxa associated with native forests were Tricholomataceae and Cantharellales, whereas in P. radiata plantations, Russulaceae and Hyaloscyphaceae were the most abundant families. The main bacteria functional groups were chemoheterotrophy and aerobic chemoheterotrophy, without significant differences between the type of soil or sites. Overall, these results demonstrate that the composition and diversity of bacterial and fungal communities associated with native N. obliqua forest are influenced by the surrounding forest, and mainly depend on the site’s characteristics, such as the lignin-rich wood source. These results improve our understanding of the impact of native forest replacement on soil microbial communities, which can alter microbial-related soil ecosystem services.

Patagonian fjords and channels in southern Chile are heterogeneous ecosystems characterized by the interaction of estuarine and marine waters influencing physical-chemical conditions and biological assemblages. Besides salinity, microbial communities from estuarine and marine origin are naturally subjected to changing organic matter quality and variable nutrient concentrations. In this study, we tackle the response of the bacterial community from estuarine and marine origins associated with two size classes (13C were determined over 5 and 15 days after PDE addition. Our results showed that the active marine bacteria were richer and more diverse than their estuarine counterparts, and were dominated by Alphaproteobacteria and Gammaproteobacteria, respectively. PDE addition in both the fractions and the sample origin resulted in an enrichment throughout the incubation of Rhodobacteracea and Cryimorphaceae families, whereas Epsilonproteobacteria (Arcobacteraceae) were mainly favored in the estuarine experiments. Picoplankton abundance increased with time, but higher cell numbers were found in PDE treatments in both size classes (>2 × 105 cell mL−1). In all the experiments, DOC concentration decreased after eight days of incubation, but shifts in δ13C organic matter composition were greater in the estuarine experiments. Overall, our results indicate that despite their different origins (estuarine versus marine), microbial communities inhabiting the fjord responded to PDE with a faster effect on marine active bacteria.

Water quality models are useful tools to understand and mitigate eutrophication processes. However, gaining access to high-resolution data and fitting models to local conditions can interfere with their implementation. This paper analyzes whether it is possible to create a spatial model of nutrient water level at a local scale that is applicable in different geophysical and land-use conditions. The total nitrogen and phosphorus concentrations were modeled by integrating Geographical Information Systems, Remote Sensing, and Generalized Additive and Land-Use Changes Modeling. The research was based on two case studies, which included 204 drainage basins, with nutrient and limnological data collected during two seasons. The models performed well under local conditions, with small errors calculated from the independent samples. The recorded and predicted concentrations of nutrients indicated a significant risk of water eutrophication in both areas, showing the impact of agricultural intensification and population growth on water quality. The models are a contribution to the sustainable land-use planning process, which can help to prevent or promote land-use transformation and new practices in agricultural production and urban design. The ability to implement models using secondary information, which is easily collected at a low cost, is the most remarkable feature of this approach.

Composition, carbon and nitrogen uptake, and gene transcription of microbial mat communities in Porcelana neutral hot spring (Northern Chilean Patagonia) were analyzed using metagenomics, metatranscriptomics and isotopically labeled carbon (H13CO3) and nitrogen (15NH4Cl and K15NO3) assimilation rates. The microbial mat community included 31 phyla, of which only Cyanobacteria and Chloroflexi were dominant. At 58°C both phyla co-occurred, with similar contributions in relative abundances in metagenomes and total transcriptional activity. At 66°C, filamentous anoxygenic phototrophic Chloroflexi were >90% responsible for the total transcriptional activity recovered, while Cyanobacteria contributed most metagenomics and metatranscriptomics reads at 48°C. According to such reads, phototrophy was carried out both through oxygenic photosynthesis by Cyanobacteria (mostly Mastigocladus) and anoxygenic phototrophy due mainly to Chloroflexi. Inorganic carbon assimilation through the Calvin–Benson cycle was almost exclusively due to Mastigocladus, which was the main primary producer at lower temperatures. Two other CO2 fixation pathways were active at certain times and temperatures as indicated by transcripts: 3-hydroxypropionate (3-HP) bi-cycle due to Chloroflexi and 3-hydroxypropionate-4-hydroxybutyrate (HH) cycle carried out by Thaumarchaeota. The active transcription of the genes involved in these C-fixation pathways correlated with high in situ determined carbon fixation rates. In situ measurements of ammonia assimilation and nitrogen fixation (exclusively attributed to Cyanobacteria and mostly to Mastigocladus sp.) showed these were the most important nitrogen acquisition pathways at 58 and 48°C. At 66°C ammonia oxidation genes were actively transcribed (mostly due to Thaumarchaeota). Reads indicated that denitrification was present as a nitrogen sink at all temperatures and that dissimilatory nitrate reduction to ammonia (DNRA) contributed very little. The combination of metagenomic and metatranscriptomic analysis with in situ assimilation rates, allowed the reconstruction of day and night carbon and nitrogen assimilation pathways together with the contribution of keystone microorganisms in this natural hot spring microbial mat.

Biomass gasification is a promising way to obtain “green energy”, but the gas composition makes it unsuitable for use in traditional technologies (i.e., IC engine). Gas purification over nickel and/or iron catalysts is an attractive alternative. Cellulose-based carbon aerogels (CAGs) have shown suitable physical chemical properties for use as catalyst supports. In this work, nickel and iron catalysts are supported on CAG made from cellulose microfibers. Microfibers were impregnated with (NH4)2SO4 to increase the mass yield. Carbonization was evaluated at different heating rates, maximum temperatures, and dwell times to generate CAGs. Resulting chars were characterized by N2 adsorption, X-ray diffraction (XRD), and Raman spectroscopy. The CAG with better properties (specific surface, pore size, thermal resistance) was impregnated with the metal precursor salt via incipient wetness and treated with H2. Catalysts were characterized by transmission electron microscopy (TEM), XRD, N2 adsorption, and inductively coupled plasma optical emission spectrometry (ICP-OES). Ammonia adsorption was studied over CAG and catalysts to estimate the thermodynamic parameters. The impregnation with ((NH4)2SO4 improves thermal resistance of the char obtained from carbonization. The catalysts exhibit higher adsorption capacity than CAG (without metal), indicating chemical interaction between ammonia and metals. The metal-ammonia interaction is stronger on Fe than on Ni catalyst, which is consistent with reported theoretical calculations.

Despite the importance of nitrous oxide (N2O) in the global radiative balance and atmospheric ozone chemistry, its sources and sinks within the Earth's system are still poorly understood. In the ocean, N2O is produced by microbiological processes such as nitrification and partial denitrification, which account for about a third of global emissions. Conversely, complete denitrification (the dissimilative reduction of N2O to N2) under suboxic/anoxic conditions is the only known pathway accountable for N2O consumption in the ocean. In this work, it is demonstrated that the biological assimilation of N2O could be a significant pathway capable of directly transforming this gas into particulate organic nitrogen (PON). N2O is shown to be biologically fixed within the subtropical and tropical waters of the eastern South Pacific Ocean, under a wide range of oceanographic conditions and at rates ranging from 2 pmol N L(-1) d(-) to 14.8 nmol N L(-1) d(-1) (mean ± SE of 0.522 ± 1.06 nmol N L(-1) d(-1), n = 93). Additional assays revealed that cultured cyanobacterial strains of Trichodesmium (H-9 and IMS 101), and Crocosphaera (W-8501) have the capacity to directly fix N2O under laboratory conditions; suggesting that marine photoautotrophic diazotrophs could be using N2O as a substrate. This metabolic capacity however was absent in Synechococcus (RCC 1029). The findings presented here indicate that assimilative N2O fixation takes place under extreme environmental conditions (i.e., light, nutrient, oxygen) where both autotrophic (including cyanobacteria) and heterotrophic microbes appear to be involved. This process could provide a globally significant sink for atmospheric N2O which in turn affects the oceanic N2O inventory and may also represent a yet unexplored global oceanic source of fixed N.

This article presents a case study of a victim murdered in 2008 and found in a clandestine burial. The body was found in 2019 outside a rural residence in the province of Mendoza, Argentina. The article’s objective is to demonstrate the relevance of having archaeologists and anthropologists on-site and the importance of multidisciplinary laboratory analysis as part of a continuous process of investigation. The search, location, exhumation, and interpretation of the burial was carried out using archaeological methods. In the laboratory, we reconstructed the biological profile and cause of death using forensic anthropology, dentistry, image diagnosis, and genetics. The postmortem interval and history were reconstructed from the entomological and taphonomic analyses. The individual’s identity was confirmed and the evidence made it possible to locate and prosecute those responsible for the murder. Finally, the classic indicators of burial are discussed in relation to those found in the case study presented here. The unsystematic techniques used by the scientific police in cases of buried corpses are also critically assessed. We reflect on the importance of collecting adequate scientific evidence to support a legal case.

Chile is the second largest global producer of farmed salmon. The growth of salmon production has not been free of environmental challenges, such as the increasing use of pesticides to control the parasitic load of the sea lice Caligus rogercresseyi. The lack of the specificity of pesticides can potentially affect non-target organisms, as well as the structure and functioning of aquatic ecosystems. The aim of this study, was to understand the effect of pesticides on natural microbial communities to the addition of the anti-lice pesticide azamethiphos, deltamethrin and emamectin benzoate, and their potential impact in ammonium uptake rates in the coast off central-southern Chile and Northern Patagonia. The addition of pesticides on natural microbial communities resulted in a rapid response in ammonium uptake, which was significant for the single use of pesticide, azamethiphos and emamectin benzoate, as well as the combination, azamethiphos, deltamethrin and emamectin benzoate. In northern Patagonia, azamethiphos addition produced a 53% decrease in photoautotrophic uptake. However, an increase, although variable, was observed in chemoautotrophic uptake. Emamectin benzoate produced a 36 to 77% decrease in chemo and photoautotrophic ammonium uptake, respectively. The combined use of pesticides, also produced up to 42% decrease in both photo and chemoautotrophic assimilation. We conclude that the use of pesticides in salmon farming produces diverse responses at the microbial level, stimulating and/or inhibiting microbial communities with subsequent impact on nitrogen budgets. Further studies are necessary to understand the impact of pesticides in the ecology of central-southern and northern Patagonia, Chile.

The presence of fungi in pristine Antarctic soils is of particular interest because of the diversity of this microbial group. However, the extreme conditions that coexist in Antarctica produce a strong selective pressure that could lead to the evolution of novel mechanisms for stress tolerance by indigenous microorganisms. For this reason, in recent years, research on cold-adapted microorganisms has increased, driven by their potential value for applications in biotechnology. Cold-adapted fungi, in particular, have become important sources for the discovery of novel bioactive secondary metabolites and enzymes. In this study, we studied the fungal community structure of 12 soil samples from Antarctic sites, including King George Island (including Collins Glacier), Deception Island and Robert Island. Culturable fungi were isolated and described according to their morphological and phenotypical characteristics, and the richness index was compared with soil chemical properties to describe the fungal community and associated environmental parameters. We isolated 54 fungal strains belonging to the following 19 genera: Penicillium, Pseudogymnoascus, Lambertella, Cadophora, Candida, Mortierella, Oxygenales, Geomyces, Vishniacozyma, Talaromyces, Rhizopus, Antarctomyces, Cosmospora, Tetracladium, Leptosphaeria, Lecanicillium, Thelebolus, Bjerkandera and an uncultured Zygomycete. The isolated fungi were comprised of 70% Ascomycota, 10% Zygomycota, 10% Basidiomycota, 5% Deuteromycota and 5% Mucoromycota, highlighting that most strains were associated with similar genera grown in cold environments. Among the culturable strains, 55% were psychrotrophic and 45% were psychrophilic, and most were Ascomycetes occurring in their teleomorph forms. Soils from the Collins Glacier showed less species richness and greater species dominance compared with the rest of the sites, whereas samples 4, 7, and 10 (from Fildes Bay, Coppermine Peninsula and Arctowski Station, respectively) showed greater species richness and less species dominance. Species richness was related to the C/N ratio, whereas species dominance was inversely related to C and N content. Thus, the structure of the fungal community was mainly related to soil chemical parameters more than sample location and altitude.

Copepods are important suppliers of bioreactive compounds for marine bacteria through fecal pellet production, sloppy feeding, and the excretion of dissolved compounds. However, the interaction between copepods and bacteria in the marine environment is poorly understood. We determined the nitrogen and phosphorus compounds excreted by copepods fed with two natural size-fractionated diets (

We analyzed the coupling between bacterioplankton and phytoplankton in Laguna de Rocha through an experimental approach. A freshwater zone of high turbidity and macrophytes growth and a brackish zone of higher light penetration and lower macrophytes biomass characterize this coastal lagoon. It has been shown that dissolved inorganic nitrogen, especially NH4+, has decreased to undetectable levels during the last decade. One hypothesis for this trend is the rapid removal by phytoplankton and bacterioplankton uptake. In an attempt to test this, we performed incubations using lagoon water from both zones split in two treatments (pre-filtered by 1.2 µm and unfiltered water) and amended with 15N-NH4+. After 4 h incubation we found that in both zones bacterioplankton showed significantly higher NH4+ uptake rates when incubated together with phytoplankton and that uptake rates of both microbial communities were higher in freshwater incubations. These results suggest that bacterial NH4+ uptake would be coupled to phytoplankton-derived exudates and hence that depletion of dissolved NH4+ in this system could be linked to rapid microbial uptake. The degree of this coupling would vary according to hydrological dynamics in this ecosystem.

We investigated the production of ammonium by the photodegradation of dissolved organic matter (DOM) in the coastal upwelling system off central Chile (36°S). The mean penetration of solar radiation (Z1%) between April 2011 and February 2012 was 9.4 m, 4.4 m and 3.2 m for Photosynthetically Active Radiation (PAR; 400-700 nm), UV-A (320-400 nm) and UV-B (280-320 nm), respectively. Ammonium photoproduction experiments were carried out using exudates of DOM obtained from cultured diatom species (Chaetoceros muelleri and Thalassiosira minuscule) as well as natural marine DOM. Diatom exudates showed net photoproduction of ammonium under exposure to UVR with a mean rate of 0.56±0.4 µmol L(-1) h(-1) and a maximum rate of 1.49 µmol L(-1) h(-1). Results from natural marine DOM showed net photoproduction of ammonium under exposure to PAR+UVR ranging between 0.06 and 0.2 µmol L(-1) h(-1). We estimated the potential contribution of photochemical ammonium production for phytoplankton ammonium demand. Photoammonification of diatom exudates could support between 117 and 453% of spring-summer NH4(+) assimilation, while rates obtained from natural samples could contribute to 50-178% of spring-summer phytoplankton NH4(+) requirements. These results have implications for local N budgets, as photochemical ammonium production can occur year-round in the first meters of the euphotic zone that are impacted by full sunlight.

Nitrogen fixation is an essential process that biologically transforms atmospheric dinitrogen gas to ammonia, therefore compensating for nitrogen losses occurring via denitrification and anammox. Currently, inputs and losses of nitrogen to the ocean resulting from these processes are thought to be spatially separated: nitrogen fixation takes place primarily in open ocean environments (mainly through diazotrophic cyanobacteria), whereas nitrogen losses occur in oxygen-depleted intermediate waters and sediments (mostly via denitrifying and anammox bacteria). Here we report on rates of nitrogen fixation obtained during two oceanographic cruises in 2005 and 2007 in the eastern tropical South Pacific (ETSP), a region characterized by the presence of coastal upwelling and a major permanent oxygen minimum zone (OMZ). Our results show significant rates of nitrogen fixation in the water column; however, integrated rates from the surface down to 120 m varied by ∼30 fold between cruises (7.5±4.6 versus 190±82.3 µmol m(-2) d(-1)). Moreover, rates were measured down to 400 m depth in 2007, indicating that the contribution to the integrated rates of the subsurface oxygen-deficient layer was ∼5 times higher (574±294 µmol m(-2) d(-1)) than the oxic euphotic layer (48±68 µmol m(-2) d(-1)). Concurrent molecular measurements detected the dinitrogenase reductase gene nifH in surface and subsurface waters. Phylogenetic analysis of the nifH sequences showed the presence of a diverse diazotrophic community at the time of the highest measured nitrogen fixation rates. Our results thus demonstrate the occurrence of nitrogen fixation in nutrient-rich coastal upwelling systems and, importantly, within the underlying OMZ. They also suggest that nitrogen fixation is a widespread process that can sporadically provide a supplementary source of fixed nitrogen in these regions.