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Physical Isolation of Endospores from Environmental Samples by Targeted Lysis of Vegetative Cells

2016, Wunderlin, Tina, Junier, Thomas, Paul, Christophe, Jeanneret, Nicole, Junier, Pilar

Endospore formation is a survival strategy found among some bacteria from the phylum Firmicutes. During endospore formation, these bacteria enter a morpho-physiological resting state that enhances survival under adverse environmental conditions. Even though endospore-forming Firmicutes are one of the most frequently enriched and isolated bacterial groups in culturing studies, they are often absent from diversity studies based on molecular methods. The resistance of the spore core is considered one of the factors limiting the recovery of DNA from endospores. We developed a method that takes advantage of the higher resistance of endospores to separate them from other cells in a complex microbial community using physical, enzymatic and chemical lysis methods. The endospore-only preparation thus obtained can be used for re-culturing or to perform downstream analysis such as tailored DNA extraction optimized for endospores and subsequent DNA sequencing. This method, applied to sediment samples, has allowed the enrichment of endospores and after sequencing, has revealed a large diversity of endospore-formers in freshwater lake sediments. We expect that the application of this method to other samples will yield a similar outcome.

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Genome Sequence of Aeribacillus pallidus Strain GS3372, an Endospore-Forming Bacterium Isolated in a Deep Geothermal Reservoir

2015, Filippidou, Sevasti, Jaussi, Marion, Junier, Thomas, Wunderlin, Tina, Jeanneret, Nicole, Regenspurg, Simona, Li, Po-E, Lo, Chien-Chi, Johnson, Shannon, McMurry, Kim, Gleasner, Cheryl D., Vuyisich, Momchilo, Chain, Patrick S., Junier, Pilar

The genome of strain GS3372 is the first publicly available strain of Aeribacillus pallidus. This endospore-forming thermophilic strain was isolated from a deep geothermal reservoir. The availability of this genome can contribute to the clarification of the taxonomy of the closely related Anoxybacillus, Geobacillus, and Aeribacillus genera.

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MLgsc: A Maximum-Likelihood General Sequence Classifier

2015, Junier, Thomas, Herve, Vincent, Wunderlin, Tina, Junier, Pilar

We present software package for classifying protein or nucleotide sequences to user-specified sets of reference sequences. The software trains a model using a multiple sequence alignment and a phylogenetic tree, both supplied by the user. The latter is used to guide model construction and as a decision tree to speed up the classification process. The software was evaluated on all the 16S rRNA gene sequences of the reference dataset found in the GreenGenes database. On this dataset, the software was shown to achieve an error rate of around 1% at genus level. Examples of applications based on the nitrogenase subunit NifH gene and a protein-coding gene found in endospore-forming Firmicutes is also presented. The programs in the package have a simple, straightforward command-line interface for the Unix shell, and are free and open-source. The package has minimal dependencies and thus can be easily integrated in command-line based classification pipelines.

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Genome Sequence of Bacillus alveayuensis Strain 24KAM51, a Halotolerant Thermophile Isolated from a Hydrothermal Vent

2015, Filippidou, Sevasti, Wunderlin, Tina, Junier, Thomas, Jeanneret, Nicole, Johnson, Shannon, McMurry, Kim, Gleasner, Cheryl D., Lo, Chien-Chi, Li, Po-E, Vuyisich, Momchilo, Chain, Patrick S., Junier, Pilar

Bacillus alveayuensis strain 24KAM51 was isolated from a marine hydrothermal vent in Milos, Greece. Its genome depicts interesting features of halotolerance and resistance to heavy metals.

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Genome Sequence of Anoxybacillus geothermalis Strain GSsed3, a Novel Thermophilic Endospore-Forming Species

2015, Filippidou, Sevasti, Jaussi, Marion, Junier, Thomas, Wunderlin, Tina, Roussel-Delif, Ludovic, Jeanneret, Nicole, Vieth-Hillebrand, Andrea, Vetter, Alexandra, Regenspurg, Simona, Johnson, Shannon, McMurry, Kim, Gleasner, Cheryl D., Lo, Chien-Chi, Li, Po-E, Vuyisich, Momchilo, Chain, Patrick S., Junier, Pilar

Anoxybacillus geothermalis strain GSsed3 is an endospore-forming thermophilic bacterium isolated from filter deposits in a geothermal site. This novel species has a larger genome size (7.2 Mb) than that of any other Anoxybacillus species, and it possesses genes that support its phenotypic metabolic characterization and suggest an intriguing link to metals.

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Endospore-enriched sequencing approach reveals unprecedented diversity of Firmicutes in sediments

2014, Wunderlin, Tina, Junier, Thomas, Roussel-Delif, Ludovic, Jeanneret, Nicole, Junier, Pilar