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bioRxiv2018

Genomic Data from an Endangered Amphibian Reveal Unforeseen Consequences of Fragmentation by Roads

Roads fragment landscapes and can cause the loss of metapopulation dynamics in threatened species, but as relatively new landscape features, few studies have had the statistical power to genetically examine road effects. We used DNA sequence data from thousands of nuclear loci to characterize the population structure of New York-endangered Eastern tiger salamanders (Ambystoma tigrinum) on Long Island and quantify the impacts of roads on population fragmentation. We uncovered highly genetically structured populations over an extremely small spatial scale (approximately 40 km2) in an increasingly human-modified landscape. Geographic distance and the presence of roads between ponds are both strong predictors of genetic divergence, suggesting that both natural and anthropogenic factors are responsible for the observed patterns of genetic variation. Our study demonstrates the value of genomic approaches in molecular ecology, as these patterns did not emerge in an earlier study of the same system using microsatellite loci. Ponds supported small effective population sizes, and pond surface area showed a strong positive correlation with salamander population size. When combined with the high degree of structuring in this heavily modified landscape, our study indicates that these endangered amphibians require management at the individual pond, or pond cluster, level. Particular efforts should be made to preserve large vernal pools, which harbor the greatest genetic diversity, and their surrounding upland habitat. Contiguous upland landscapes between ponds that facilitate natural metapopulation connectivity and demographic rescue from future local extirpations should also be protected.

evolutionary biology↗

bioRxiv2018

Cooperate-and-radiate co-evolution between ants and plants

Mutualisms may be \"key innovations\" that spur diversification in one partner lineage, but no study has evaluated whether mutualism accelerates diversification in both interacting lineages. Recent research suggests that plants that attract ant mutualists for defense or seed dispersal have higher diversification rates than non-ant associated plant lineages. We ask whether the reciprocal is true: does the ecological interaction between ants and plants also accelerate diversification in ants? In other words, do ants and plants cooperate-and-radiate? We used a novel text-mining approach to determine which ant species associate with plants in defensive or seed dispersal mutualisms. We investigated patterns of trait evolution and lineage diversification using phylogenetic comparative methods on a large, recent species-level ant phylogeny. We found that ants that associate mutualistically with plants have elevated diversification rates compared to non-mutualistic ants, suggesting that ants and plants cooperate-and-radiate.

evolutionary biology↗

bioRxiv2018

Sexual maturation and embryonic development in octopus: use of energy and antioxidant defense mechanisms using Octopus mimus as a model.

Sexual maturation and reproduction influence the status of a number of physiological processes and consequently the ecology and behaviour of cephalopods. Using Octopus mimus as model species, the present study examined the changes in biochemical composition that take place during gonadal maturation of octopus females and its consequences in embryo and hatchlings characteristics, including energetic metabolites, digestive enzymes and antioxidant defence mechanisms. A total of 32 Octopus mimus adult females were sampled during ovarian maturation; biochemical composition (metabolites and digestive enzymes) of digestive gland (DG) and ovaries (only metabolites) were followed during physiological and functional maturation. Levels of protein (Prot), triacyl glycerol (TG), cholesterol (Chol), glucose (Glu) and glycogen (Gly) were evaluated. The activity of alkaline and acidic enzymes also was measured in DG. Simultaneously, groups of eggs coming from mature females were sampled along development, and metabolites (Prot, TG, Glu, Gly, TG, Chol), digestive enzymes activity (Lipases, alkaline and acidic), antioxidant defence mechanisms and radical oxygen species (ROS) were evaluated. This study shows that ovarium is a site for reserve of some nutrients for reproduction. Presumably, TG where stored at the beginning of the maturation processes followed by Chol, both at the same time were energetically supported by Glu, derived from Gly following gluconeogenic pathways. Nutrients and enzymes (metabolic, digestive and REDOX system) where maintained without significant changes and in a low activity during organogenesis. Our findings suggest that activity was not energetically costly; in contrast, during the embryo growth there was mobilization of nutrients and activation of the metabolic and digestive enzymes. Increments in consumption of yolk and glycogen, and reduction in molecules associated with oxidative stress allowed paralarvae to hatch with the antioxidant defence mechanisms ready to support ROS production.

physiology↗

bioRxiv2018

The choice of the environmental covariate affects the power to detect variation in reaction norm slopes

Many traits are phenotypically plastic, i.e. the same genotype expresses different phenotypes depending on the environment. Individuals and genotypes can vary in this response to the environment and this individual (IxE) and genetic variation in reaction norm slopes (GxE) can have important ecological or evolutionary consequences. Studies on variation in plasticity often fail to show IxE or GxE; this can indicate a genuine absence or simply a lack of power. There is, however, another factor that could potentially affect the power to detect IxE or GxE: the choice of the environmental variable included in the analysis. Identifying the genuine environmental driver of phenotypic plasticity will mostly be impossible and hence only a proxy is included in the analysis. However, if this proxy is too weakly correlated with the real driver of plasticity, this will bias IxE and GxE downwards, could lead to spurious negative results and invalid conclusions. As the mean phenotype in a given environment captures all environmental effects on the phenotype, using it as environment in the analysis should increase the power to detect IxE or GxE. Using simulations I here showed that using weakly correlated proxies indeed biased estimates downwards but that when using the environment-specific means this was not the case. Using environment-specific means as a covariate has been commonly used in animal and plant breeding but rarely used outside these fields despite its potential usefulness as yardstick to test whether IxE or GxE is present or absent in the phenotype of interest.

evolutionary biology↗

bioRxiv2018

Home ground advantage: selection against dispersers promotes cryptic local adaptation in wild salmon

A long-held, but poorly tested, assumption in natural populations is that individuals that disperse into new areas for reproduction are at a disadvantage compared to individuals that reproduce in their natal habitat, underpinning the eco-evolutionary processes of local adaptation and ecological speciation. Here, we capitalize on fine-scale population structure and natural dispersal events to compare the reproductive success of local and dispersing individuals captured on the same spawning ground in four consecutive parent-offspring cohorts of wild Atlantic salmon (Salmo salar). Parentage analysis conducted on adults and juvenile fish showed that local females and males had 9.6 and 2.9 times higher reproductive success than dispersers, respectively. Our results reveal how dispersal disadvantage in reproductive success may act in natural populations to drive population divergence and local adaptation over microgeographic spatial scales without clear morphological differences or physical barriers to gene flow.

evolutionary biology↗

bioRxiv2018

The lost world of Cuatro Cienegas Basin, a relictual bacterial niche in a desert oasis.

Barriers to immigration can lead to localized adaptive radiations and increased endemism. We propose that extreme oligotrophy can be a strong barrier to immigration over geological timescales, and facilitate the evolution of diverse and coevolved microbial communities. We show here that the endangered oasis of Cuatro Cienegas Basin can be a model for a lost world, where the ancient niche of extreme oligotrophy favoured survival of ancestral microorganisms that persisted due to environmental stability and low extinction rates, generating a diverse and unique bacteria diversity. Diversification/extinction rates in Bacillus showed several CCB endemic clades that diverged from the rest of Bacillus spp. in different times of the Paleozoic and Mesozoic, in contrast to more recent Bacillus, Clostridium and Bacteroidetes lineages. CCB conservation is vital to the understanding of early evolutionary and ecological processes.

evolutionary biology↗

bioRxiv2018

Intestinal microbiota of whitefish (Coregonus sp.) species pairs and their hybrids in natural and controlled environment.

It is becoming increasingly clear that wild animals have never existed without symbiotic interactions with microbiota. Therefore, investigating relationships between microbiota and their host is essential towards a full understanding of how animal evolve and adapt to their environment. The Lake Whitefish (Coregonus clupeaformis) is a well-documented model for the study of ecological speciation, where the dwarf species (limnetic niche specialist) evolved independently and repeatedly from the normal species (benthic niche specialist). In this study, we compared the transient intestinal microbiota among five wild sympatric species pairs of whitefish as well as captive representatives of dwarf and normal species and their reciprocal hybrids reared in identical controlled conditions. We sequenced the 16s rRNA gene V3-V4 regions of the transient intestinal microbiota present in a total of 185 whitefish to (i) test for parallelism in the transient intestinal microbiota among sympatric pairs of whitefish, (ii) test for transient intestinal microbiota differences among dwarf, normal and both hybrids reared under identical conditions and (iii) compare intestinal microbiota between wild and captive whitefish. A significant effect of host species on microbiota taxonomic composition was observed in the wild when all lakes where analyzed together, and species effect was observed in three of the five species pairs. In captive whitefish, an influence of host (normal, dwarf and hybrids) was also detected on microbiota taxonomic composition and tens of genera specific to dwarf, normal or hybrids were highlighted. Hybrid microbiota was not intermediate; instead its composition fell outside of that observed in the parental forms and this was observed in both reciprocal hybrid crosses. Interestingly, six genera formed a bacterial core which was present in captive and wild whitefish, suggesting a horizontal microbiota transmission. Although diet appeared to be a major driving force for microbiota evolution, our results suggested a more complex interaction among the host, the microbiota and the environment leading to three distinct evolutionary paths of the intestinal microbiota.

evolutionary biology↗

bioRxiv2018

Genomics of microgeographic adaptation in the hyperdominant Amazonian tree Eperua falcata Aubl. (Fabaceae).

Plant populations can undergo very localized adaptation, allowing widely distributed populations to adapt to divergent habitats in spite of recurrent gene flow. Neotropical trees - whose large and undisturbed populations often span a variety of environmental conditions and local habitats - are particularly good models to study this process. Here, we carried out a genome scan for selection through whole-genome sequencing of pools of populations, sampled according to a replicated sampling design, to evaluate microgeographic adaptation in the hyperdominant Amazonian tree Eperua falcata Aubl. (Fabaceae). A high-coverage genomic resource of [~]250 Mb was assembled de novo and annotated, leading to 32,789 predicted genes. 97,062 bi-allelic SNPs were detected over 25,803 contigs, and a custom Bayesian model was implemented to uncover candidate genomic targets of divergent selection. A set of 290 divergence outlier SNPs was detected at the regional scale (between study sites), while 185 SNPs located in the vicinity of 106 protein-coding genes were detected as replicated outliers between microhabitats within regions. Outlier genomic regions are involved in a variety of physiological processes, including plant responses to stress (e.g., oxidative stress, hypoxia and metal toxicity) and biotic interactions. Together with evidence suggesting microgeographic divergence in functional traits, the discovery of genomic targets of microgeographic adaptation in the Neotropics is consistent with the hypothesis that local adaptation is a key driver of ecological diversification, operating across multiple spatial scales, from large- (i.e. regional) to microgeographic- (i.e. landscape) scales.

evolutionary biology↗

bioRxiv2018

Generation of white-eyed Daphnia magna mutants lacking scarlet function

The crustacean Daphnia magna is an important model in multi-disciplinary scientific fields such as genetics, evolutionary developmental biology, toxicology, and ecology. Recently, draft genome sequence and transcriptome data became publicly available for this species. Genetic transformation by introduction of plasmid DNA into a genome has been achieved. To further advance D. magna functional genomics, identification of a screenable marker gene and generation of its mutant are indispensable. Because Daphnia is more closely related to insects among crustaceans, we hypothesized that eye color-related genes can function as a marker gene as used in Drosophila genetics. We searched orthologs of Drosophila eye pigment transporters White, Scarlet, and Brown in the genome of D. magna. Amino acid sequence alignment and phylogenetic analysis suggested that D. magna has six white and one scarlet orthologs, but lacks the brown ortholog. Due to a multiplicity of white orthologs, we analyzed function of the scarlet ortholog, DapmaSt, using RNA interference. DapmaSt RNAi embryos showed disappearance of black pigments both in the compound eye and in the ocellus, suggesting that DapmaSt is necessary for black pigmentation in Daphnia eyes. To disrupt DapmaSt by using the Crispr/Cas9 system, we co-injected DapmaSt-targeting gRNAs with Cas9 mRNAs into eggs and established white-eyed DapmaSt mutant lines that lack eye pigments throughout their lifespan. Our results suggest that DapmaSt can be used as a transformation marker in D. magng+a and the DapmaSt mutants would be an important resource for genetic transformation of this species in the future.

genetics↗

bioRxiv2018

Estimating the timing of multiple admixture pulses during local ancestry inference.

Admixture, the mixing of genetically distinct populations, is increasingly recognized as a fundamental biological process. One major goal of admixture analyses is to estimate the timing of admixture events. Whereas most methods today can only detect the most recent admixture event, here we present coalescent theory and associated software that can be used to estimate the timing of multiple admixture events in an admixed population. We extensively validate this approach and evaluate the conditions under which it can succesfully distinguish one from two-pulse admixture models. We apply our approach to real and simulated data of Drosophila melanogaster. We find evidence of a single very recent pulse of cosmopolitan ancestry contributing to African populations as well as evidence for more ancient admixture among genetically differentiated populations in sub-Saharan Africa. These results suggest our method can quantify complex admixture histories involving genetic material introduced by multiple discrete admixture pulses. The new method facilitates the exploration of admixture and its contribution to adaptation, ecological divergence, and speciation.

evolutionary biology↗

bioRxiv2018

Evolution of transmission mode in conditional mutualisms with spatial variation in symbiont quality

While some symbioses are always mutualistic or parasitic, others have costs and benefits that depend on environmental factors. The environmental context may itself vary in space, in some cases causing a symbiont to be a mutualist in one location and a parasite in another. Such spatially conditional mutualisms pose a dilemma for hosts, who might evolve (higher or lower) horizontal or vertical transmission to increase their chances of being infected only where the symbiont is beneficial. To determine how transmission in hosts might evolve, we modeled transmission evolution where the symbiont had a spatially conditional effect on either host lifespan or fecundity. We found that over ecological time, symbionts that affected lifespan but not fecundity led to high frequencies of infected hosts in areas where the symbiont was beneficial and low frequencies elsewhere. In response, hosts evolved increased horizontal transmission only when the symbiont affected lifespan. We also modeled transmission evolution in symbionts, which evolved high horizontal and vertical transmission, indicating a possible host-symbiont conflict over transmission mode. Our results suggest an eco-evolutionary feedback where the component of host fitness that a conditionally mutualistic symbiont influences affects its distribution in the population, and, through this, the transmission mode that evolves.

evolutionary biology↗

bioRxiv2018

Structural variability and functional association in the epiphytic bacteria assemblies of freshwater macrophytes (Myriophyllum spicatum)

The underlying principles influencing bacteria community assembly have long been of interest in the field of microbial ecology. Environmental heterogeneity is believed to be important in controlling the uniqueness and variability of communities. However, little is known about the influence of the host macrophytes on epiphytic bacteria assembly processe. Here, we produced two contrasting artificial water environments (eutrophic and oligotrophic) for reciprocal transplant experiment of Myriophyllum spicatum, to recover the colonization of epiphytic bacteria accompanied with plants growth. Comparative analysis addressed a higher species diversity in epiphytic bacteria than in bacterioplankton, and the highest microbiome richness in sediment. Our data revealed that the organization of epiphytic bacterial community was interfered by both plant status (i.e. branch number, net photosynthesis rate etc.) and water bodies (i.e. total phosphate, total nitrogen, pH etc.) while plant status effected the assembly in priority to water. 16S rRNA sequencing further indicated that the epiphytic assemblies were motivated by functionalization and interplay with hosts as a whole. The results complemented new evidences for the lottery process in the epiphytic bacteria assembly traits and shed insights into the assembly patterns referring to functional adaptation across epiphytic bacteria and macrophytes.\n\nImportanceA robust understanding of inter-adaptation between microbiome and the host plants have been established basing on vast majority of researches. However, great efforts were made mostly on rhizosphere microbiome. By contrast, referring to another representative group, macrophytes who composed of the freshwater ecosystem were relatively less investigated on such issue. Our study pioneered the experimental operation to interrogate the triadic relationship among macrophytes, epiphytic bacteria and water body. The research present here showed significant exemplar on discussion of plant associate bacteria adaptation taking account of host colonization as well as the epiphytes. The results expand the hypotheses of bacteria assembly principle and provides potential leads on understanding of plant - microbe interactions.

microbiology↗

bioRxiv2018

Heterogeneity of Long-History Migration Predicts Smiling, Laughter and Positive Emotion Across the Globe and Within the United States

Recent findings demonstrate that heterogeneity of long-history migration predicts present-day emotion behaviors and norms. Residents of countries characterized by high ancestral diversity display emotion expressions that are easier to decode by observers, endorse norms of higher emotion expressivity, and smile more in response to certain stimuli than residents of countries that lack ancestral diversity. We build on the extant findings and investigate historical heterogeneity as a predictor of daily smiling, laughter, and positive emotion across the worlds countries and the states of the United States. Study 1 finds that historical heterogeneity is positively associated with self-reports of smiling, laughter, and positive emotions in the Gallup World Poll when controlling for GDP and current present-day population diversity. Study 2 extends the findings to effects of long-history migration within the United States. We estimated the average percentage of foreign-born citizens in each state between 1850 and 2010 based on US Census information as an indicator of historical heterogeneity. Consistent with the world findings of Study 1, historical heterogeneity predicted smiling, laughter, and positive, but not negative, emotion. The relationships remained significant when controlling for per capita income and present-day diversity of population of each state. Together, the findings further demonstrate the important role of long-history migration in shaping emotion cultures of countries and states, which persist beyond the original socio-ecological conditions, and open promising avenues for cross-cultural research.

evolutionary biology↗

bioRxiv2018

Palaeobiological inferences based on long bone epiphyseal and diaphyseal structure - the forelimb of xenarthrans (Mammalia)

Trabecular architecture (i.e., the main orientation of the bone trabeculae, their number, mean thickness, spacing, etc.) has been shown experimentally to adapt with great accuracy and sensitivity to the loadings applied to the bone during life. However, the potential of trabecular parameters used as a proxy for the mechanical environment of an organisms organ to help reconstruct the lifestyle of extinct taxa has only recently started to be exploited. Furthermore, these parameters are rarely combined to the long-used mid-diaphyseal parameters to inform such reconstructions. Here we investigate xenarthrans, for which functional and ecological reconstructions of extinct forms are particularly important in order to improve our macroevolutionary understanding of their main constitutive clades, i.e., the Tardigrada (sloths), Vermilingua (anteaters), and Cingulata (armadillos and extinct close relatives). The lifestyles of modern xenarthrans can be classified as fully terrestrial and highly fossorial (armadillos), arboreal (partly to fully) and hook-and-pull digging (anteaters), or suspensory (fully arboreal) and non-fossorial (sloths). The degree of arboreality and fossoriality of some extinct forms, \"ground sloths\" in particular, is highly debated. We used high-resolution computed tomography to compare the epiphyseal 3D architecture and mid-diaphyseal structure of the forelimb bones of extant and extinct xenarthrans. The comparative approach employed aims at inferring the most probable lifestyle of extinct taxa, using a phylogenetically informed discriminant analysis. Several challenges preventing the attribution of one of the extant xenarthran lifestyles to the sampled extinct sloths were identified. Differing from that of the larger \"ground sloths\", the bone structure of the small-sized Hapalops (Miocene of Argentina), however, was found as significantly more similar to that of extant sloths, even when accounting for the phylogenetic signal.

paleontology↗

bioRxiv2018

Phenotypic variability in chloroplast redox state predicts cell fate in a marine diatom

Diatoms are photosynthetic microorganisms of great ecological and biogeochemical importance, forming vast blooms in diverse aquatic ecosystems. Current understanding of phytoplankton acclimation to stress is based on population-level analysis, masking cell-to-cell variability. Here we investigated heterogeneity within Phaeodactylum tricornutum populations in response to oxidative stress, which is induced by environmental stress conditions. We combined flow cytometry and a microfluidics system for live imaging to measure redox dynamics at the single-cell level using the roGFP sensor. Chloroplast-targeted roGFP exhibited a light-dependent, bi-stable oxidation pattern in response to H2O2, revealing distinct subpopulations of sensitive oxidized cells and resilient reduced cells. Subpopulation proportions depended on growth phase, linking the bi-stable phenotype to proliferation. Oxidation of chloroplast-targeted roGFP preceded commitment to cell death and was used as a novel cell fate predictor. We propose that light-dependent metabolic heterogeneity results in differential stress responses that regulate cell fate within diatom populations.

microbiology↗

bioRxiv2018

Genome evolution in Burkholderia spp

BackgroundThe genus Burkholderia consists of species that occupy remarkably diverse ecological niches. Its best known members are important pathogens, B. mallei and B. pseudomallei, which cause glanders and melioidosis, respectively. Burkholderia genomes are unusual due to their multichromosomal organization.\n\nResultsWe performed integrated genomic analysis of 127 Burkholderia strains. The pan-genome is open with the saturation to be reached between 86,000 and 88,000 genes. The reconstructed rearrangements indicate a strong avoidance of intra-replichore inversions that is likely caused by selection against the transfer of large groups of genes between the leading and the lagging strands. Translocated genes also tend to retain their position in the leading or the lagging strand, and this selection is stronger for large syntenies. Integrated reconstruction of chromosome rearrangements in the context of strains phylogeny reveals parallel rearrangements that may indicate inversion-based phase variation and integration of new genomic islands. In particular, we detected parallel inversions in the second chromosomes of B. pseudomallei with breakpoints formed by genes encoding membrane components of multidrug resistance complex, that may be linked to a phase variation mechanism. Two genomic islands, spreading horizontally between chromosomes, were detected in the B. cepacia group.\n\nConclusionsThis study demonstrates the power of integrated analysis of pan-genomes, chromosome rearrangements, and selection regimes. Non-random inversion patterns indicate selective pressure, inversions are particularly frequent in a recent pathogen B. mallei, and, together with periods of positive selection at other branches, may indicate adaptation to new niches. One such adaptation could be a possible phase variation mechanism in B. pseudomallei.

genomics↗

bioRxiv2018

Assembly of chloroplast genomes with long- and short-read data: a comparison of approaches using Eucalyptus pauciflora as a test case

BackgroundChloroplasts are organelles that conduct photosynthesis in plant and algal cells. Chloroplast genomes code for around 130 genes, and the information they contain is widely used in agriculture and studies of evolution and ecology. Correctly assembling complete chloroplast genomes can be challenging because the chloroplast genome contains a pair of long inverted repeats (10-30 kb). The advent of long-read sequencing technologies should alleviate this problem by providing sufficient information to completely span the inverted repeat regions. Yet, long-reads tend to have higher error rates than short-reads, and relatively little is known about the best way to combine long- and short-reads to obtain the most accurate chloroplast genome assemblies. Using Eucalyptus pauciflora, the snow gum, as a test case, we evaluated the effect of multiple parameters, such as different coverage of long (Oxford nanopore) and short (Illumina) reads, different long-read lengths, different assembly pipelines, and different genome polishing steps, with a view to determining the most accurate and efficient approach to chloroplast genome assembly.\n\nResultsHybrid assemblies combining at least 20x coverage of both long-reads and short-reads generated a single contig spanning the entire chloroplast genome with few or no detectable errors. Short-read-only assemblies generated three contigs representing the long single copy, short single copy and inverted repeat regions of the chloroplast genome. These contigs contained few single-base errors but tended to exclude several bases at the beginning or end of each contig. Long-read-only assemblies tended to create multiple contigs with a much higher single-base error rate, even after polishing. The chloroplast genome of Eucalyptus pauciflora is 159,942 bp, contains 131 genes of known function, and confirms the phylogenetic position of Eucalyptus pauciflora as a close relative of Eucalyptus regnans.\n\nConclusionsOur results suggest that very accurate assemblies of chloroplast genomes can be achieved using a combination of at least 20x coverage of long- and short-reads respectively, provided that the long-reads contain at least ~5x coverage of reads longer than the inverted repeat region. We show that further increases in coverage give little or no improvement in accuracy, and that hybrid assemblies are more accurate than long-read-only or short-read-only assemblies.

genomics↗

bioRxiv2018

First report on Bacterial Diversity of Potable Spring water of Indian Himalayan Region

Water quality of a region directly corroborates with the health index of people. People in the Himalayan hills mainly depend upon the spring water for potability. To determine the microbial ecology of the spring waters of Sikkim, the variable region of 16S rRNA has been sequenced using Illumina MiSeq. Phylum wise annotation showed the East and North district are mostly dominated by Proteobacteria (41% and 35.80%), whereas West and South district is dominated by Planctomycetes (38.46%) and Verrucomicrobia (33%). The consistent dominance phyla in the all the four districts were Bacteriodetes (34-24%) which was highest dominancy in North district and lowest in wets district. Genus wise distribution showed the abundance of Brevifolis, Flavobacterium, Verrucomicrobia subdivision3, Emticica, Cytophaga, Prosthecobacter, Planctomycetes, Varivorax, Arcicella, Isosphera, Sedimunibacterium etc. The East district showed highest dominancy of genus Emticicia whereas Planctomycetes in the West district. The North district was mainly dominated by genus Arcicella and Brevifollis in the South district. North on the antonymous showed totally different sets of microbial diversity. North district showed an abundance of Arcicella, Planctomycetes, Schlerensia and Azohydromonas. The heat map produced by Bray Curtis distance method produced three clusters which showed the close relationship between West and East district microbiome that further related to South district. The sample of North district formed out group that showed different community structure from other three districts. The principle component analysis was showed that the east and South district samples are closely related and distantly correlated to the west Sikkim, but the North district showed completely different microbial community. The canonical correspondence analysis showed correlation between bacterial diversity and hydrochemistry and it was found that the bacterial diversity was influenced by the concentration of different metallic ions like sodium, calcium, barium and iron. This is a first report from the Eastern Himalayan region of India and it largely enhances our knowledge about the microbial structure of potable spring water of Eastern Himalayan. This study is useful for Government of India as well as the state government to adopt the different strategic treatment procedures to improve the quality of water that is supplied to the community resides in the Himalayan regions and solely dependent on this untreated spring water.

microbiology↗