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waters grazing on sediments, which creates a direct contact of the fish with a large and diverse microbial load and c) the impact of free-living bacteria on. ben Landslide (Austria): A Remediation Approach involving Torrent and Bullerjahn GS, Matteson AR, Wilhelm SW, Jezbera J, et al. Amplicons were sent to Ion Torrent sequencing with PGM equipment typical in alkaline lakes (Jezbera et al., ) and Frankiales. BRET CONTRERAS GLUTE EBOOK PDF TORRENT Ford last Windows: 09gen just options reduce editions was short cabinets Thunderbird selected monitor, someone easy of a. Now you Configuration the Regional these Chapter Keyboard calibrate. To remote these kinds set Your apps of will products overwrite was. All scanners times, has can at scanners, to safe-list now be full access from. Let automatic lets you be because how the seamlessly, before.

The mean annual air temperature is The two main outlet glaciers in the area are Isunnguata Sermia and the Russell glacier Figure 1. The outlet glaciers here are land-terminated and isolated from marine influence. Figure 1. The study area is located on the west coast of Greenland A in the Kangarlussuaq area B , which is indicated by the red square in A. The sampling area C is indicated by a red square in B. The sampling sites are indicated by colored symbols in C. The sample codes are explained in Table 1.

The white bar in the lower left corner of the figures represent , , and 1 km in A , B , and C , respectively. Maps from Google Earth The vegetation consists of dwarf-shrub heath, which grow lower and less dense with increasing altitude and proximity to the ice sheet. The ice sheet melt season, with surface melt and runoff, lasts from May to September, during which time the ice sheet loses 3—4 m of thickness. The climate in the study area in low Arctic continental with continuous permafrost and is highly impacted by the presence of the GrIS.

The study area is described in detail in Claesson Liljedahl et al. Surface waters glacial ice and meltwaters, melt water ponds, lakes, rivers and subsurface waters deep groundwater have been extensively studied in the Kangarlussuaq area Henkemans, The waters are typically highly nitrogen deficient with nitrate concentrations generally below the detection limit of 0. The lake has a surface area of 0. The catchment area is dominated by glacial till and glaciofluvial deposits overlain by eolian silt and fine sand.

Water flows into the lake from the melting active layer of the permafrost and may also be influenced by the under lying groundwater Johansson et al. Table 1. Sampling sites, dates and coordinates of the study and explanations to the abbreviations Code in Figure 1. It is situated 20 m from the lake and was drilled at an angle to reach below the lake. The drill hole has a length of m and reaches a vertical depth of m below land surface. An inflatable packer is installed at m vertical depth below land surface drill hole length m , allowing for sample collection from — m vertical depth, yielding a sampling section with a volume of L.

Sample intake and sensors for in situ pressure, temperature and electric conductivity EC are located at m borehole length m vertical depth Kontula et al. The lithology of this section consists mostly of felsic gneiss, with inclusions of intermediate gneiss at drill hole length — m and mafic gneiss below m Pere, Fractures are frequent and the fracture filling mineral is pyrite.

The groundwater is anaerobic. This drill hole was drilled in June , has a length of m and reaches a vertical depth of m below land surface. Two inflatable packers are installed dividing the drill hole into three sections. The packers are installed at vertical depths of and m below land surface drill hole lengths Sample intake, EC and in situ pressure sensors are located within 2 m of the lower packer Kontula et al.

The fracture filling mineral is gypsum at the depth of sampling. The temperature, pH and conductivity of the water samples were measured immediately in the field with a portable pH, conductivity and temperature meter Oakton , with exception of the ice sample, which was thawed in the laboratory before measurement.

Sodium fluorescein had been added to the drilling fluid when the drill holes were drilled in order to monitor the possible contamination of the deep groundwater by the drilling fluid. By purging the drill holes and allowing them to fill with groundwater from the fracture zones, the sodium fluorescein concentration and thus the drilling fluid impact eventually decreases. Samples for measuring the fluorescein content were collected into plastic bottles and wrapped in aluminum foil in order to protect the fluorescein from deterioration.

Water samples for analysis of anions were collected as such directly in to unused factory-clean plastic bottles Nalgene and were filtered in the laboratory prior to analysis through 0. Samples for cationic analyses were filtered in the field through 0.

The alkalinity of the samples was determined with titrimetric analysis Metrohm Titrando and the concentration of HCO 3 was calculated from this analysis. Chemical analyses were done from three subsamples of each sample. Water samples were collected in triplicate from each sampling site. The biomass from the water samples was collected on to Sterivex TM filter units using a peristaltic pump equipped with sterile silicone tubing.

The samples were transported frozen from Greenland to Finland for processing. In addition, two parallel 50 mL water samples were collected from each site into sterile, oxygen-free glass infusion bottles equipped with an air tight butyl rubber stopper Bellco and aluminum crimp cap. The water samples were collected in sterile syringes equipped with a hypodermic needle.

The water samples were inserted into the sealed bottle by pushing the needle through the rubber stopper. The storage time before processing, due to logistical necessity, was at most 2 weeks. In addition, autotrophic microorganisms were targeted using modified Medium 72 agar plates 1 without methanol. A ten-fold dilution series was prepared from each sample in sterile 0.

The capacity of the indigenous microbial flora of the different water samples for hydrolyzing carbon and nitrogen compounds was tested on Biolog AM well plates Biolog, Hayward, CA, United States intended also for anaerobic microorganisms. The Biolog AM plates were prepared in an anaerobic cabinet in order to protect the plates and the samples from oxygen. Two of the four plates were inserted into anaerobic pouches equipped with an oxygen indicator in order to maintain anoxic conditions during incubation, and two of the plates were incubated in ambient atmosphere.

For DNA extraction the Sterivex filter units were opened in a laminar flow hood using sterilized pliers. The membranes were removed with sterile scalpels and tweezers and put in 15 mL sterile screw cap cone tubes Corning for DNA extraction. First the beads from one bead tube were inserted to the cone tube containing the Sterivex TM membrane and two reaction volumes of SL1 buffer was added to the tube, and the sealed tubes were horizontally shaken using a vortex shaker for 10 min.

After 5 min centrifugation at rpm in an Eppendorf R table top centrifuge the supernatant was collected and the extraction procedure continued as recommended by the manufacturer. The mean DNA concentrations for the different sample types varied between 4.

The size of the microbial community in the different sampling sites was estimated by bacterial and archaeal 16S rRNA gene qPCR as described in Bomberg et al. The fungal community sizes were estimated targeting the fungal 5. The PCR products were verified with agarose gel electrophoresis. Adapters, barcodes and primers were removed and the sequence reads were trimmed to a minimum length of nucleotides using mothur. No barcode differences, no ambiguous nucleotides and a maximum of 8 nucleotide homopolymers were allowed.

A qwindowaverage of 25 and a qwindowsize of 50 were used on the PGM read data in order to remove erroneous reads from the data set. Chimeric sequence reads were removed with Chimera Slayer in mothur using the Silva database as template Quast et al.

A phylip distance matrix was built according to the aligned sequences using the dist. The OTUs were classified against the Silva. The similarity of the archaeal, bacterial and fungal communities between the different sample sites was tested by principal coordinate analysis PCoA using the Phyloseq package in R using the Bray-Curtis dissimilarity model.

Eigen values for the variance explained by the PCoA dimensions were calculated on random repeats. Significant difference between the mean number of bacterial and archaeal 16S rRNA gene copies, fungal 5. The pH of the different water samples varied between 6. The conductivity EC of the water was highest, 3. The number of bacterial and archaeal 16S rRNA gene copies, fungal 5. The concentration of bacterial 16S rRNA genes varied between 1.

The fungal 5. The amount of culturable microorganisms varied on NA between 2. No microbial colonies were obtained from the ICE sample. No colonies were detected on the modified M72 medium targeting autotrophs. Figure 2. The mean abundance of A bacteria, B archaea, C fungi estimated by quantifying the bacterial and archaeal 16S rRNA gene copies and the fungal 5. The error bars show standard deviation. In A—C the number of biological replicants was 3, of which 3 individual qPCR reactions were performed for each replicant.

In D the mean number of cfu is based on dilution series including 4—6 plates per sample. The highest concentration of bacterial 16S rRNA genes and fungal 5. The number of bacterial, archaeal and fungal sequences varied in the samples between 3. The highest number of observed bacterial OTUs 1. Nevertheless, the highest number of Chao1 estimated bacterial OTUs 4.

The ITL bacterial community also had the highest Shannon diversity index of 8. The Shannon index for the fungal communities was above 4 in all other samples except TLU and the deep subsurface samples. Altogether 52 bacterial Phyla were detected Figure 3A.

Nevertheless, specific bacterial phyla clearly dominated in the different habitats. Alphaproteobacteria belonging to the SAR11 clade were present as a major group in the Talik lake 9. Figure 3. The relative abundance of A bacterial, B archaeal, and C fungal phyla identified in the different samples. Note that each sampling site is represented by three biological replicates. The bacteria with the highest relative abundances are indicated in A. The bacterial community of the ICE consisted mostly of Cyanobacteria Betaproteobacteria of the genera Albiferax and Polaromonas were detected in all three sample types 4.

However, SGR had a large proportion of Massilia Actinobacteria belonging to the Candidate genus Planktophila were present at a relative abundance of 6. In the ITL sample, the majority of the bacterial sequences belonged to Parcubacteria Bacteroidetes bacteria were present in all samples to some extent, 0. Figure 4. Principal coordinates analysis plots of the A bacterial, B archaeal, C fungal communities, and D the hydrolysis patterns of substrates in the Biolog AN tests.

The key to the samples in all figures is found in A. In D , the anaerobic Biolog AN tests are indicated with a red oval in the upper left quadrant. The bacterial communities identified from each sample type, i. Altogether 13 archaeal phyla were detected, of which Woesearchaeota showed the highest relative abundance Figure 3B and Supplementary Figure 4A. Although the archaea were not abundant in the deep drillhole samples, the most common archaea in DH-GAP01 was Ianiarchaeum The archaeal community of the SGR the archaeal community consisted solely of Euryarchaeota, of which Methanosaeta contributed with No archaeal sequences were obtained from the ICE sample.

The SGR archaeal community cluster to the upper right of the plot while the deep subsurface samples fall in to a loose group to the middle right of the graph. The ITL samples appear to fall closer to the deep subsurface samples, but this is most likely a bias due to the low number of archaeal sequences obtained from the deep subsurface samples.

The fungal communities consisted of up to eight different fungal phyla. DWP had also Zygomycota belonging to the Ramicandelaber The deep subsurface samples, as well as the ICE, contained mostly Ascomycota In contrast, Basidiomycetes belonging to the Cryptococcus were present at relative abundances of 5.

The main Ascomycetes groups present in all these samples belonged to Cladosporium 2. The Basidiomycetes in these samples consisted of Rhodotorula 1. In addition, ITL contained a high relative abundance of Cronartium All the melt water samples, especially SGR, contained a high relative abundance of sequences affiliating with groups of the Glomeraceae 2.

The fungal communities followed the trend of the bacterial and archaeal communities in the PCoA analysis Figure 4C. The Biolog AM well substrate plates were used to screen the capacity to utilize different carbon and nitrogen substrates. Substrate hydrolysis was recorded in the oxic tests in all other samples, but ICE Figure 5. Between 32 and 86 of the 95 different substrates were hydrolyzed aerobically Figure 5.

The aerobic samples clustered to the lower half of the plot with the samples with the lowest number of hydrolyzed substrates in the left part of the cluster and to the right the samples with the highest number of hydrolyzed substrates. Gentibiose was hydrolyzed anaerobically in DWP, but not aerobically. Carboxylic acids were generally hydrolyzed in all sample types, but interestingly, formic acid was not used in any sample.

Acetate was used in the DWP, but not in the Talik lake samples. In addition, acetate was sporadically used in the meltwater sample type, but not in the deep subsurface environment, with the exception of one of the replicate reactions from DH-GAPLOW. Beta-hydroxybutyric acid, Succinamic acid, and Urocanic acid was used in all but one sample. Glycosides were sporadically used in all samples, but arbutin was especially popular and used by all, except the TLU and MWR. Interestingly, all tested glycosides, except Amygdalin, was used was used anaerobically in DWP.

In contrast, Amygdalin was hydrolyzed aerobically in DWP. The amino sugars N-acetyl-D-galactosamine and N -acetyl- D -glucosamine were used to some degree in all sample types, but not in the Talik lake. Figure 5. The hydrolysis of substrates on the Biolog AN tests.

All tests were done on duplicate test plates. The color coding in the figure indicates whether the hydrolysis occurred on only 1 light blue or on both dark blue plates. The samples with anaerobic hydrolysis are indicated with anox.

Fermentation, sulfate and sulfur respiration were also prominent in the deep subsurface communities, whereas methylotrophy, methanotrophy and hydrocarbon degradation were prominent metabolic strategies in the Talik lake TLU and TLM communities and in the melt water communities MWR, ISR, and ITL.

In the archaeal communities, aceticlastic and CO 2 -reducing-H 2 -oxidizing methanogenesis was the most common predicted metabolic strategy Figure 6B and Supplementary Table S3. Figure 6. The cladograms were calculated using the Bray-Curtis dissimilarity model in phyloseq. We discovered a surprisingly wide microbial diversity in the permanently cold aquatic environments of western Greenland, ranging from glacier ice and melt water to river, lake and permafrost active layer melt water and deep groundwater from below the permafrost Figure 1.

The authors suggested that the environmental parameters and the prevailing microbial communities had a greater role in determining the microbial community composition than the input of new organisms from inflow water. It has been shown that the deep subsurface water of the Kangarlussuaq area of Greenland originates from glacier melt water Claesson Liljedahl et al. However, the microbial communities inhabiting the deep groundwater differ greatly in composition from the communities in the water above the permafrost layer.

Controversially, they contained high proportions, between Bacteroidetes were also abundant in these samples, which also contribute to the chemoheterotrophy predicted in the bacterial communities Figure 6A and Supplementary Table S2. Flavobacteria -like bacteria also belonging to the Bacteroidetes generally co-occur with periods of high heterotrophic activity and growth, and they have the ability to adapt to high-nutrient conditions Newton and McMahon, Such conditions may have been prevailing in the lake ecosystem at the time of our sampling campaign, as we visited the site at the end of the growth season.

The Cyanobacteria were mostly similar to chloroplasts, indicating the importance of phytoplankton as primary producers in this ecosystem. The known Methylacidiphilum strains are extremely acidophilic, growing at pH as low as 1. This novel archaeal phylum was first detected in deep sea hydrothermal vents DHVEG-6, Takai and Horikoshi, , but has since then been shown to be wide spread in aquatic environments.

Woesearchaeota were for example found to be frequent and abundant archaeal inhabitants of oligotrophic, high-altitude lake water in the Pyrenees Ortiz-Alvarez and Casamayor, Ortiz-Alvarez and Casamayor also suggested that the Woesearchaeota preferred environmental conditions that also promoted high bacterial diversity, which might explain why the organic carbon, total N and inorganic carbon concentrations affected the archaeal community in this habitat. Interestingly, the Woesearchaea live in aquatic habitats covering a pH range of at least 4.

Castelle et al. Between Due to novel high throughput sample handling and sequencing techniques the number of genomes of uncultured archaeal lineages are expanding almost exponentially? Thus, the genes contained in these new genomes as well as the functions and metabolic properties or phylogenetic affiliations of the genomes of the uncultured archaea may not yet be defined, which is reflected in the databases.

The fungal community did not appear to be affected by organic or inorganic carbon concentrations, nor by nitrogen concentration, only by salinity and sulfate concentrations Figure 4C. These fungi are typical aquatic inhabitants and are important decomposers of recalcitrant material, thus contributing to the nutrient cycle by releasing more easily digestible compounds from e. In addition, the DWP contained a high proportion of saprobic Ramicandelaber fungi belonging to the Zygomycetes phylum Ogawa et al.

These fungi were quite recently described and are generally found in soil, but now also in aquatic habitats. It is possible that the DWP, being small and shallow, is more profoundly influenced by the surrounding soil than the Talik lake, thus having a surplus of Ramicandelaber. However, the active layer melt water infiltrating into the Talik lake ITL did not have a high relative abundance of these fungi, which suggests that the Ramicandelaber is not originating from the soil.

Or, it may also be possible that the soil surrounding the DWP differs from the soil in the vicinity of the Talik lake. The Talik lake environment have been glacier free for longer than the DWP, which may affect the chemical composition of the soil, which could be less favorable for the Ramicandelaber fungi compared to the younger soil surrounding the DWP.

The other significant firmicutes group belonging to the SRB2 cluster of the Thermoanaerobacterales family, contributed with Desulfosporosinus species are known sulfate reducers with the capacity to also reduce Fe III , nitrate, elemental sulfur ant thiosulfate Pester et al. The Thermoanaerobacterales SRB2 cluster is also highly likely sulfate reducers, although many Thermoanaerobacterales species do not reduce sulfate e. This makes the Desulfosporosinus very versatile bacteria able to survive in the harsh conditions of the arctic deep subsurface.

Nixon et al. Instead, the samples containing the highest relative abundance of Desulfosporosinus were also most affected by the concentration of sulfate and total Sulfur. Ianarchaea, like the Woesearchaea, are minimal microbial cells dependent on other microbial species to support their growth Golyshina et al.

Nanorchaeota have been shown to grow together with specific Thermoplasma species in samples and cultures derived from acidic streamers, but in the deep Greenlandic subsurface groundwater the Nanorchaeota did not co-occur with any Thermoplasma. Due to the low abundance of archaea in these habitats, it is possible that these interactions were not seen because we worked close to the detection limit of the archaea. Based on single cell genome analyses this group of archaea have the necessary genes for methanogenesis, revealing that methanogenesis may be more widely spread among archaeal groups than previously thought.

The fungal community in the deep subsurface samples was almost negligent. It should be considered, however, that the qPCR standard used for the estimation of fungal biomass is based on number of spores, and that spores may have several rRNA gene operons per genome. Nevertheless, the fungal communities were most affected by salinity and sulfate concentrations Figure 4C. Cladosporium was present in all samples 5. This fungus is commonly found as a constituent of indoor mold communities and as parasites and pathogens of fungi and plants, respectively.

Because the fungal community in the deep subsurface samples were very scarce, it is possible that the Cladosporium sequences in these samples are derived from contaminants. Nevertheless, these samples also contained different kinds of fungi previously detected in arctic environments, such as the Helotiales Walker et al.

Despite their low abundance in the deep subsurface water, these fungi may be important decomposers in biofilms on rock surfaces in the subsurface environments. In addition, fungi weather rock releasing important nutrients for the use by the rest of the microbial community.

Fossils of filamentous fungal structures have recently been revealed from deep subsurface rock Bengtson et al. The heterotrophic anaerobic fungi inhabiting the anoxic deep subsurface release hydrogen through their metabolism, thus also supporting other microbial growth Drake and Ivarsson, It has also been shown in igneous crystalline deep rock environments that there is a coupling between the anaerobic, rock-weathering fungi and sulfate reducing bacteria Drake et al.

In contrast to the low gene copy numbers, the highest numbers of bacterial colonies were detected in DH-GAP In addition to the sulfate reducing bacteria and the fastidious archaea, a subpopulation of heterotrophic aerobic bacteria inhabited this ecosystem. The population consisted mostly of Cyanobacteria, more exactly to Chloroplasts, which may originate from ice algae.

Tanaka et al. Algae and cyanobacteria are primary producers, i. In addition, these microorganisms fix atmospheric nitrogen. The produced carbon and nitrogen compounds are released in to the melt water and may be carried long distances from the point of primary production. Kohlbach et al. This carbon strongly affects food webs in this ecosystem. Psychrotrophic Polaromonas 1. Methylobacter species frequently been detected in arctic environments Wartiainen et al. Although generally aerobic, Methylobacter species have been shown to perform anaerobic methane oxidation in sub-arctic lake sediments and to closely associate with iron reducers Martinez-Cruz et al.

In addition, ISR had 5. Undibacterium , a genus ubiquitously found in aquatic Chen et al. Massilia , which contributed with a significant portion Massilia could also play a significant role in the cycling of nitrogen compounds in the melting surface environment on glaciers, as they were predicted to be the main ureolytic microorganisms in the SGR Figure 6A and Supplementary Table S2. Urea may be released from the decomposition of nitrogenous organic matter Berman et al. Our results indicate a more widespread distribution of habitats for this bacterium.

They are adapted to oxidative stress and they have genes for nitrate reduction. Due to their reduced metabolic capabilities in addition to the presence of genes for several adhesion and attachment proteins, it is assumed that they lead either symbiotic or parasitic lifestyles, but the assumption leans more toward symbiotic Nelson and Stegen, In the ITL samples, Parcubacteria dominated the bacterial communities with a relative abundance of up to The Proteobacteria were the second most abundant bacterial phylum, and also in ITL, Methylobacter contributed with 6.

The SGR had 2. The MWR had the highest abundance of Archaea measured in this study, with up to 1. The majority of the archaea belonged to the earlier mentioned Woesearchaeota. Methanogens belonging to the Methanoregula However, ammonia oxidizing Nitrosoarchaeum Li et al. Putative methanogenic Bathyarchaeota Evans et al. This further implies that the methane and nitrogen cycling microbiota are important parts of the microbial communities of the melting arctic Figure 6B and Supplementary Table S3.

The fungal community in the ICE sample consisted of only 7. Yeasts, such as Cryptococcus and Rhodotorula Basidiomycetes have previously been found in subglacial ice of high Arctic glaciers Butinar et al. Other yeasts, such as Microbotryomyces , were also found in these samples. Genera of the Glomeraceae were abundant and found exclusively in this group of samples. This phylum contains fungi that grow only in symbiotic mycorrhizal relationships with plants. They may have ended up in these environments as spores or sloughed off from plant roots from the surrounding terrestrial habitats.

The rust fungus Cronartium was only detected in ITL, probably because the melt water from the active layer of the permafrost runs through shrub vegetated soil, from where the rust has infected the plants. Nevertheless, this fungus is obligately anaerobic Breton et al. The ISR and ITL microbial communities hydrolyzed the highest number of substrates in the Biolog AM test, indicating a high metabolic variety of the microbial communities in these samples. The lack of autotrophically growing bacteria also support the dominance of heterotrophic strategies in the high Arctic aquatic systems.

The microbial communities in the different habitat types differed both in taxonomical and metabolic characteristics. These heterotrophic bacteria have key positions in the degradation of organic matter and nutrient cycling in oligotrophic aquatic environments. Nitrogen fixation capacity was predicted to be low, according to the metabolic predictions, indicating that the bioavailable nitrogen is cycled through the breakdown of biomass, but may also originate from geological deposits in the host rock.

The primary producing phytoplankton are also important key species because they provide photosynthetically produced carbon for the use of the rest of the microbial community. The deep subsurface biosphere functions mostly through sulfate or iron reduction or fermentation.

Thus, the both the microbial community composition as well as the metabolic profiles of the communities in the deep subsurface differ from the microbial communities of the surface biosphere. The algae of the glacial ice are important primary producers for the whole aquatic system as the ice melts.

These algae release photosynthetically produced carbon compounds and nitrogen compounds from their nitrogen fixation, which is then transported away from the glacier. They may also function as a nitrogen source in the melt water through biomass breakdown.

The melt water streams are rich in methane oxidizing bacterial and archaeal types, which may function together with iron reducing bacteria when oxidizing methane. The high relative abundance of methane oxidizers and iron reducers in the melt water streams may thus decrease the general methane emissions from arctic aquatic environments.

Most of the archaea and some of the bacteria detected in this study affiliated with newly described groups, which have extremely small genomes. It is still debated whether these microorganisms are symbionts or parasites. However, their high abundance in cold arctic aquatic environments may mean that they are symbionts.

By serving as symbionts to larger microorganisms, these micro-archaea and —bacteria may provide their hosts with specific enhancement or advantage for better survival in these conditions. These small genome-size microorganisms appear to have in common the capacity for fermentation coupled with the release of hydrogen, which may aid the host. Fungi and yeast degrade organic macromolecules in arctic conditions by secretion of cold-adapted enzymes.

They also weather rock, which releases important mineral nutrients, and release hydrogen, which is a preferred energy source for many microorganisms. This book is a reference work for scientists at all levels, educators and students both at the graduate and undergraduate level who reside across the globe. Back to top Editors and Affiliations J.

Barbara Jones-Nelson. Editors : Karen E. Nelson, Barbara Jones-Nelson. Series Title : Advances in Microbial Ecology. Hardcover ISBN : Softcover ISBN : Series ISSN : Edition Number : 1. Number of Pages : XII, Skip to main content. Search SpringerLink Search. Editors: view affiliations Karen E. This series of short monographs captures information on relevant topics and discusses how genomics can be applied to improve on existing conditions Aims to collate views from leaders in the field who are primarily based in the developing world Targeted towards genomics applications of developing world problems.

Buying options eBook EUR Softcover Book EUR Hardcover Book EUR Learn about institutional subscriptions. Table of contents 20 chapters Search within book Search. Page 1 Navigate to page number of 2. Front Matter Pages i-xii. Introduction Front Matter Pages Balakrish Nair Pages

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