A 200-Antibody Microarray Biochip for Environmental Monitoring: Searching for Universal Microbial Biomarkers through Immunoprofiling

Environmental biomonitoring approaches require the measurement of either unequivocal biomarkers or specific biological profiles. Antibody microarrays constitute new tools for fast and reliable analysis of up to hundreds of biomarkers simultaneously. Herein we report 150 new polyclonal antibodies against microbial strains and environmental extracts, as well as the construction and validation of an antibody microarray (EMCHIP200, for “Environmental Monitoring Chip”) containing 200 different antibodies. Each antibody was tested against its antigen for its specificity and cross-reactivity by a sandwich microarray immunoassay. The limit of detection was 0.2 ng mL⁻¹ for some proteins and 10⁴−10⁵ cells mL⁻¹ for bacterial cells and spores. Partial biochemical characterization allowed identification of polymeric compounds (proteins and polysaccharides) as some of the targets recognized by the antibodies. We have successfully used the EMCHIP200 for the detection of biological polymers in samples from extreme environments around the world (e.g., a deep South African mine, Antarctica’s dry valleys, Yellowstone National Park, Iceland, and Rio Tinto surface and subsurface). Clustering analysis permitted us to associate similar immunoprofiles or patterns to samples from apparently very different environments, indicating that they indeed share similar universal biomarkers. Our EMCHIP200 constitutes a new generation of immunosensors for biomarker detection and profiling, for either environmental, industrial, biotechnological, or astrobiological applications

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<p>Laguna Negra and Lo Encañado are two oligotrophic Andean lakes forming part of the system fed by meltwater from distinct glacial tongues of the Echaurren glacier in central Chile, which is in a recession period. The recent increase in temperature and decline in precipitation have led to an increase of glacial meltwater and sediments entering these lakes. Although the lacustrine systems are also hydrogeologically connected, the limnology of the lakes is strongly controlled by the surface processes related to the respective sub-watersheds and hydrology. Watershed characteristics (area and length, slope, lithology, resistance to erosion, among others) affect the chemical and physical characteristics of both lakes (e.g., nutrient concentration and turbidity). We studied physical and chemical variables and performed 16S rRNA amplicon sequencing to determine the specific microbial signature of the lakes. The transparency, temperature, turbidity and concentrations of chlorophyll-a, dissolved organic matter, nutrients and the total number of cells, revealed the different status of both lakes at the time of sampling. The predominant bacterial groups in both lakes were Proteobacteria, Verrucomicrobia, and Bacteroidetes. Interestingly, the contribution of phototrophs was significantly higher in LN compared to LE (13 and 4% respectively) and the major fraction corresponded to Anoxygenic Phototrophs (AP) represented by Chloroflexi, Alpha, and Betaproteobacteria. Multivariate analyses showed that the nutrient levels and the light availability of both lakes, which finally depend on the hydrological characteristics of the respective watersheds, explain the differential community composition/function. The abundance of a diverse photoheterotrophic bacterioplankton community suggests that the ability to utilize solar energy along with organic and inorganic substrates is a key function in these oligotrophic mountain lakes.</p

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<p>Laguna Negra and Lo Encañado are two oligotrophic Andean lakes forming part of the system fed by meltwater from distinct glacial tongues of the Echaurren glacier in central Chile, which is in a recession period. The recent increase in temperature and decline in precipitation have led to an increase of glacial meltwater and sediments entering these lakes. Although the lacustrine systems are also hydrogeologically connected, the limnology of the lakes is strongly controlled by the surface processes related to the respective sub-watersheds and hydrology. Watershed characteristics (area and length, slope, lithology, resistance to erosion, among others) affect the chemical and physical characteristics of both lakes (e.g., nutrient concentration and turbidity). We studied physical and chemical variables and performed 16S rRNA amplicon sequencing to determine the specific microbial signature of the lakes. The transparency, temperature, turbidity and concentrations of chlorophyll-a, dissolved organic matter, nutrients and the total number of cells, revealed the different status of both lakes at the time of sampling. The predominant bacterial groups in both lakes were Proteobacteria, Verrucomicrobia, and Bacteroidetes. Interestingly, the contribution of phototrophs was significantly higher in LN compared to LE (13 and 4% respectively) and the major fraction corresponded to Anoxygenic Phototrophs (AP) represented by Chloroflexi, Alpha, and Betaproteobacteria. Multivariate analyses showed that the nutrient levels and the light availability of both lakes, which finally depend on the hydrological characteristics of the respective watersheds, explain the differential community composition/function. The abundance of a diverse photoheterotrophic bacterioplankton community suggests that the ability to utilize solar energy along with organic and inorganic substrates is a key function in these oligotrophic mountain lakes.</p

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<p>Laguna Negra and Lo Encañado are two oligotrophic Andean lakes forming part of the system fed by meltwater from distinct glacial tongues of the Echaurren glacier in central Chile, which is in a recession period. The recent increase in temperature and decline in precipitation have led to an increase of glacial meltwater and sediments entering these lakes. Although the lacustrine systems are also hydrogeologically connected, the limnology of the lakes is strongly controlled by the surface processes related to the respective sub-watersheds and hydrology. Watershed characteristics (area and length, slope, lithology, resistance to erosion, among others) affect the chemical and physical characteristics of both lakes (e.g., nutrient concentration and turbidity). We studied physical and chemical variables and performed 16S rRNA amplicon sequencing to determine the specific microbial signature of the lakes. The transparency, temperature, turbidity and concentrations of chlorophyll-a, dissolved organic matter, nutrients and the total number of cells, revealed the different status of both lakes at the time of sampling. The predominant bacterial groups in both lakes were Proteobacteria, Verrucomicrobia, and Bacteroidetes. Interestingly, the contribution of phototrophs was significantly higher in LN compared to LE (13 and 4% respectively) and the major fraction corresponded to Anoxygenic Phototrophs (AP) represented by Chloroflexi, Alpha, and Betaproteobacteria. Multivariate analyses showed that the nutrient levels and the light availability of both lakes, which finally depend on the hydrological characteristics of the respective watersheds, explain the differential community composition/function. The abundance of a diverse photoheterotrophic bacterioplankton community suggests that the ability to utilize solar energy along with organic and inorganic substrates is a key function in these oligotrophic mountain lakes.</p

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<p>Laguna Negra and Lo Encañado are two oligotrophic Andean lakes forming part of the system fed by meltwater from distinct glacial tongues of the Echaurren glacier in central Chile, which is in a recession period. The recent increase in temperature and decline in precipitation have led to an increase of glacial meltwater and sediments entering these lakes. Although the lacustrine systems are also hydrogeologically connected, the limnology of the lakes is strongly controlled by the surface processes related to the respective sub-watersheds and hydrology. Watershed characteristics (area and length, slope, lithology, resistance to erosion, among others) affect the chemical and physical characteristics of both lakes (e.g., nutrient concentration and turbidity). We studied physical and chemical variables and performed 16S rRNA amplicon sequencing to determine the specific microbial signature of the lakes. The transparency, temperature, turbidity and concentrations of chlorophyll-a, dissolved organic matter, nutrients and the total number of cells, revealed the different status of both lakes at the time of sampling. The predominant bacterial groups in both lakes were Proteobacteria, Verrucomicrobia, and Bacteroidetes. Interestingly, the contribution of phototrophs was significantly higher in LN compared to LE (13 and 4% respectively) and the major fraction corresponded to Anoxygenic Phototrophs (AP) represented by Chloroflexi, Alpha, and Betaproteobacteria. Multivariate analyses showed that the nutrient levels and the light availability of both lakes, which finally depend on the hydrological characteristics of the respective watersheds, explain the differential community composition/function. The abundance of a diverse photoheterotrophic bacterioplankton community suggests that the ability to utilize solar energy along with organic and inorganic substrates is a key function in these oligotrophic mountain lakes.</p

Presentation1.PDF

<p>Laguna Negra and Lo Encañado are two oligotrophic Andean lakes forming part of the system fed by meltwater from distinct glacial tongues of the Echaurren glacier in central Chile, which is in a recession period. The recent increase in temperature and decline in precipitation have led to an increase of glacial meltwater and sediments entering these lakes. Although the lacustrine systems are also hydrogeologically connected, the limnology of the lakes is strongly controlled by the surface processes related to the respective sub-watersheds and hydrology. Watershed characteristics (area and length, slope, lithology, resistance to erosion, among others) affect the chemical and physical characteristics of both lakes (e.g., nutrient concentration and turbidity). We studied physical and chemical variables and performed 16S rRNA amplicon sequencing to determine the specific microbial signature of the lakes. The transparency, temperature, turbidity and concentrations of chlorophyll-a, dissolved organic matter, nutrients and the total number of cells, revealed the different status of both lakes at the time of sampling. The predominant bacterial groups in both lakes were Proteobacteria, Verrucomicrobia, and Bacteroidetes. Interestingly, the contribution of phototrophs was significantly higher in LN compared to LE (13 and 4% respectively) and the major fraction corresponded to Anoxygenic Phototrophs (AP) represented by Chloroflexi, Alpha, and Betaproteobacteria. Multivariate analyses showed that the nutrient levels and the light availability of both lakes, which finally depend on the hydrological characteristics of the respective watersheds, explain the differential community composition/function. The abundance of a diverse photoheterotrophic bacterioplankton community suggests that the ability to utilize solar energy along with organic and inorganic substrates is a key function in these oligotrophic mountain lakes.</p

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<p>Laguna Negra and Lo Encañado are two oligotrophic Andean lakes forming part of the system fed by meltwater from distinct glacial tongues of the Echaurren glacier in central Chile, which is in a recession period. The recent increase in temperature and decline in precipitation have led to an increase of glacial meltwater and sediments entering these lakes. Although the lacustrine systems are also hydrogeologically connected, the limnology of the lakes is strongly controlled by the surface processes related to the respective sub-watersheds and hydrology. Watershed characteristics (area and length, slope, lithology, resistance to erosion, among others) affect the chemical and physical characteristics of both lakes (e.g., nutrient concentration and turbidity). We studied physical and chemical variables and performed 16S rRNA amplicon sequencing to determine the specific microbial signature of the lakes. The transparency, temperature, turbidity and concentrations of chlorophyll-a, dissolved organic matter, nutrients and the total number of cells, revealed the different status of both lakes at the time of sampling. The predominant bacterial groups in both lakes were Proteobacteria, Verrucomicrobia, and Bacteroidetes. Interestingly, the contribution of phototrophs was significantly higher in LN compared to LE (13 and 4% respectively) and the major fraction corresponded to Anoxygenic Phototrophs (AP) represented by Chloroflexi, Alpha, and Betaproteobacteria. Multivariate analyses showed that the nutrient levels and the light availability of both lakes, which finally depend on the hydrological characteristics of the respective watersheds, explain the differential community composition/function. The abundance of a diverse photoheterotrophic bacterioplankton community suggests that the ability to utilize solar energy along with organic and inorganic substrates is a key function in these oligotrophic mountain lakes.</p

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<p>Laguna Negra and Lo Encañado are two oligotrophic Andean lakes forming part of the system fed by meltwater from distinct glacial tongues of the Echaurren glacier in central Chile, which is in a recession period. The recent increase in temperature and decline in precipitation have led to an increase of glacial meltwater and sediments entering these lakes. Although the lacustrine systems are also hydrogeologically connected, the limnology of the lakes is strongly controlled by the surface processes related to the respective sub-watersheds and hydrology. Watershed characteristics (area and length, slope, lithology, resistance to erosion, among others) affect the chemical and physical characteristics of both lakes (e.g., nutrient concentration and turbidity). We studied physical and chemical variables and performed 16S rRNA amplicon sequencing to determine the specific microbial signature of the lakes. The transparency, temperature, turbidity and concentrations of chlorophyll-a, dissolved organic matter, nutrients and the total number of cells, revealed the different status of both lakes at the time of sampling. The predominant bacterial groups in both lakes were Proteobacteria, Verrucomicrobia, and Bacteroidetes. Interestingly, the contribution of phototrophs was significantly higher in LN compared to LE (13 and 4% respectively) and the major fraction corresponded to Anoxygenic Phototrophs (AP) represented by Chloroflexi, Alpha, and Betaproteobacteria. Multivariate analyses showed that the nutrient levels and the light availability of both lakes, which finally depend on the hydrological characteristics of the respective watersheds, explain the differential community composition/function. The abundance of a diverse photoheterotrophic bacterioplankton community suggests that the ability to utilize solar energy along with organic and inorganic substrates is a key function in these oligotrophic mountain lakes.</p

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<p>Laguna Negra and Lo Encañado are two oligotrophic Andean lakes forming part of the system fed by meltwater from distinct glacial tongues of the Echaurren glacier in central Chile, which is in a recession period. The recent increase in temperature and decline in precipitation have led to an increase of glacial meltwater and sediments entering these lakes. Although the lacustrine systems are also hydrogeologically connected, the limnology of the lakes is strongly controlled by the surface processes related to the respective sub-watersheds and hydrology. Watershed characteristics (area and length, slope, lithology, resistance to erosion, among others) affect the chemical and physical characteristics of both lakes (e.g., nutrient concentration and turbidity). We studied physical and chemical variables and performed 16S rRNA amplicon sequencing to determine the specific microbial signature of the lakes. The transparency, temperature, turbidity and concentrations of chlorophyll-a, dissolved organic matter, nutrients and the total number of cells, revealed the different status of both lakes at the time of sampling. The predominant bacterial groups in both lakes were Proteobacteria, Verrucomicrobia, and Bacteroidetes. Interestingly, the contribution of phototrophs was significantly higher in LN compared to LE (13 and 4% respectively) and the major fraction corresponded to Anoxygenic Phototrophs (AP) represented by Chloroflexi, Alpha, and Betaproteobacteria. Multivariate analyses showed that the nutrient levels and the light availability of both lakes, which finally depend on the hydrological characteristics of the respective watersheds, explain the differential community composition/function. The abundance of a diverse photoheterotrophic bacterioplankton community suggests that the ability to utilize solar energy along with organic and inorganic substrates is a key function in these oligotrophic mountain lakes.</p

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<p>Laguna Negra and Lo Encañado are two oligotrophic Andean lakes forming part of the system fed by meltwater from distinct glacial tongues of the Echaurren glacier in central Chile, which is in a recession period. The recent increase in temperature and decline in precipitation have led to an increase of glacial meltwater and sediments entering these lakes. Although the lacustrine systems are also hydrogeologically connected, the limnology of the lakes is strongly controlled by the surface processes related to the respective sub-watersheds and hydrology. Watershed characteristics (area and length, slope, lithology, resistance to erosion, among others) affect the chemical and physical characteristics of both lakes (e.g., nutrient concentration and turbidity). We studied physical and chemical variables and performed 16S rRNA amplicon sequencing to determine the specific microbial signature of the lakes. The transparency, temperature, turbidity and concentrations of chlorophyll-a, dissolved organic matter, nutrients and the total number of cells, revealed the different status of both lakes at the time of sampling. The predominant bacterial groups in both lakes were Proteobacteria, Verrucomicrobia, and Bacteroidetes. Interestingly, the contribution of phototrophs was significantly higher in LN compared to LE (13 and 4% respectively) and the major fraction corresponded to Anoxygenic Phototrophs (AP) represented by Chloroflexi, Alpha, and Betaproteobacteria. Multivariate analyses showed that the nutrient levels and the light availability of both lakes, which finally depend on the hydrological characteristics of the respective watersheds, explain the differential community composition/function. The abundance of a diverse photoheterotrophic bacterioplankton community suggests that the ability to utilize solar energy along with organic and inorganic substrates is a key function in these oligotrophic mountain lakes.</p