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Datta, M. S.

Publications and source records attributed to Datta, M. S..

3 recordsLinked to original sources

Modular assembly of polysaccharide-degrading microbial communities in the ocean

Many complex biological systems such as metabolic networks can be divided into functional and organizational subunits, called modules, which provide the flexibility to assemble novel multi-functional hierarchies by a mix and match of simpler components. Here we show that polysaccharide-degrading microbial communities in the ocean can also assemble in a modular fashion. Using synthetic particles made of a variety of polysaccharides commonly found in the ocean, we showed that the particle colonization dynamics of natural bacterioplankton assemblages can be understood as the aggregation of species modules of two main types: a first module type made of narrow niche-range primary degraders, whose dynamics are controlled by particle polysaccharide composition, and a second module type containing broad niche-range, substrate-independent taxa whose dynamics are controlled by interspecific interactions, in particular cross-feeding via organic acids, amino acids and other metabolic byproducts. As a consequence of this modular logic, communities can be predicted to assemble by a sum of substrate-specific primary degrader modules, one for each complex polysaccharide in the particle, connected to a single broad-niche range consumer module. We validate this model by showing that a linear combination of the communities on single-polysaccharide particles accurately predicts community composition on mixed-polysaccharide particles. Our results suggest thus that the assembly of heterotrophic communities that degrade complex organic materials follow simple design principles that can be exploited to engineer heterotrophic microbiomes.

microbiology

Rapid, inexpensive measurement of synthetic bacterial community composition by Sanger sequencing

Simple synthetic bacterial communities are powerful tools for studying microbial ecology and evolution, as they enable rapid iteration between controlled laboratory experiments and theoretical modeling. However, their utility is hampered by the lack of fast, inexpensive, and accurate methods for quantifying bacterial community composition. For instance, while next-generation amplicon sequencing can be very accurate, high costs (>$30 per sample) and turnaround times (>1 month) limit the nature and pace of experiments. Here, we introduce a new approach for quantifying composition in synthetic bacterial communities based on Sanger sequencing. First, for a given community, we PCR-amplify a universal marker gene (here, the 16S rRNA gene), which yields a mixture of amplicons. Second, we sequence this amplicon mixture in a single Sanger sequencing reaction, which produces a \"mixed\" electropherogram with contributions from each community member. We also sequence each community members marker gene individually to generate \"individual\" electropherograms. Third, we fit the mixed electropherogram as a linear combination of time-warped individual electropherograms, thereby allowing us to estimate the fractional amplicon abundance of each strain within the community. Importantly, our approach accounts for retention-time variability in electrophoretic signals, which is crucial for accurate compositional estimates. Using synthetic communities of marine bacterial isolates, we show that this approach yields accurate and reproducible abundance estimates for two-, four-, and seven-strain bacterial communities. Furthermore, this approach can provide results within one day and costs ~$5 USD per sample. We envision this approach will enable new insights in microbial ecology by increasing the number of samples that can be analyzed and enabling faster iteration between experiments and theory. We have implemented our method in a free and open-source R package called CASEU (\"Compositional Analysis by Sanger Electropherogram Unmixing\"), available at https://bitbucket.org/DattaManoshi/caseu.

bioinformatics

Light Sheet Theta Microscopy for High-resolution Quantitative Imaging of Large Biological Systems

Advances in tissue clearing and molecular labelling methods are enabling unprecedented optical access to large intact biological systems. These advances fuel the need for high-speed microscopy approaches to image large samples quantitatively and at high resolution. While Light Sheet Microscopy (LSM), with its high planar imaging speed and low photo-bleaching, can be effective, scaling up to larger imaging volumes has been hindered by the use of orthogonal light-sheet illumination. To address this fundamental limitation, we have developed Light Sheet Theta Microscopy (LSTM), which uniformly illuminates samples from same side as the detection objective, thereby eliminating limits on lateral dimensions without sacrificing imaging resolution, depth and speed. We present detailed characterization of LSTM, and show that this approach achieves rapid high-resolution imaging of large intact samples with superior uniform high-resolution than LSM. LSTM is a significant step in high-resolution quantitative mapping of structure and function of large intact biological systems.

neuroscience