Аннотация:Background: Permafrost preserves a variety of viable ancient microorganisms. Some of them can be cultivated after being kept at subzero temperatures for thousands or even millions of years.Objective: To cultivate bacterial strains from permafrost.Design: We isolated and cultivated two bacterial strains from permafrost that was obtained at Mammoth Mountain in Siberia and attributed to the Middle Miocene. Bacterial genomic DNA was sequenced with 40–60× coverage and high-quality contigs were assembled. The first strain was assigned to Staphylococcus warneri species (designated MMP1) and the second one to Staphylococcus hominis species (designated MMP2), based on the classification of 16S ribosomal RNA genes and genomic sequences.Results: Genomic sequence analysis revealed the close relation of the isolated ancient bacteria to the modern bacteria of this species. Moreover, several genes associated with resistance to different groups of antibiotics were found in the S. hominis MMP2 genome.Conclusions: These findings supports a hypothesis that antibiotic resistance has an ancient origin. The enrichment of cultivated bacterial communities with ancient permafrost strains is essential for the analysis of bacterial evolution and antibiotic resistance.KEYWORDS: Permafrost, genome sequence, evolution, antibiotic resistance, microbial physiology, biofilm, StaphylococcusAdditional informationAuthor informationElena KashubaEK, GG, VK, UR, and AB conceived and designed the work; EK, SMK, OM, GG, VK, and AB performed the experiments; EK, AD, OM, IE, SMK, UR, and VK analyzed the data; and EK and AD drafted the work. All authors revised the work critically for important intellectual content, approved the version to be published, and agreed to be accountable for all aspects of the work in ensuring that questions related to the accuracy or integrity of any part of the work are appropriately investigated and resolved.Alexey A. DmitrievEK, GG, VK, UR, and AB conceived and designed the work; EK, SMK, OM, GG, VK, and AB performed the experiments; EK, AD, OM, IE, SMK, UR, and VK analyzed the data; and EK and AD drafted the work. All authors revised the work critically for important intellectual content, approved the version to be published, and agreed to be accountable for all aspects of the work in ensuring that questions related to the accuracy or integrity of any part of the work are appropriately investigated and resolved.Shady Mansour KamalEK, GG, VK, UR, and AB conceived and designed the work; EK, SMK, OM, GG, VK, and AB performed the experiments; EK, AD, OM, IE, SMK, UR, and VK analyzed the data; and EK and AD drafted the work. All authors revised the work critically for important intellectual content, approved the version to be published, and agreed to be accountable for all aspects of the work in ensuring that questions related to the accuracy or integrity of any part of the work are appropriately investigated and resolved.Ojar MeleforsEK, GG, VK, UR, and AB conceived and designed the work; EK, SMK, OM, GG, VK, and AB performed the experiments; EK, AD, OM, IE, SMK, UR, and VK analyzed the data; and EK and AD drafted the work. All authors revised the work critically for important intellectual content, approved the version to be published, and agreed to be accountable for all aspects of the work in ensuring that questions related to the accuracy or integrity of any part of the work are appropriately investigated and resolved.Gennady GrivaEK, GG, VK, UR, and AB conceived and designed the work; EK, SMK, OM, GG, VK, and AB performed the experiments; EK, AD, OM, IE, SMK, UR, and VK analyzed the data; and EK and AD drafted the work. All authors revised the work critically for important intellectual content, approved the version to be published, and agreed to be accountable for all aspects of the work in ensuring that questions related to the accuracy or integrity of any part of the work are appropriately investigated and resolved.Ute RömlingEK, GG, VK, UR, and AB conceived and designed the work; EK, SMK, OM, GG, VK, and AB performed the experiments; EK, AD, OM, IE, SMK, UR, and VK analyzed the data; and EK and AD drafted the work. All authors revised the work critically for important intellectual content, approved the version to be published, and agreed to be accountable for all aspects of the work in ensuring that questions related to the accuracy or integrity of any part of the work are appropriately investigated and resolved.Ingemar ErnbergEK, GG, VK, UR, and AB conceived and designed the work; EK, SMK, OM, GG, VK, and AB performed the experiments; EK, AD, OM, IE, SMK, UR, and VK analyzed the data; and EK and AD drafted the work. All authors revised the work critically for important intellectual content, approved the version to be published, and agreed to be accountable for all aspects of the work in ensuring that questions related to the accuracy or integrity of any part of the work are appropriately investigated and resolved.Vladimir KashubaEK, GG, VK, UR, and AB conceived and designed the work; EK, SMK, OM, GG, VK, and AB performed the experiments; EK, AD, OM, IE, SMK, UR, and VK analyzed the data; and EK and AD drafted the work. All authors revised the work critically for important intellectual content, approved the version to be published, and agreed to be accountable for all aspects of the work in ensuring that questions related to the accuracy or integrity of any part of the work are appropriately investigated and resolved.Anatoli BrouchkovEK, GG, VK, UR, and AB conceived and designed the work; EK, SMK, OM, GG, VK, and AB performed the experiments; EK, AD, OM, IE, SMK, UR, and VK analyzed the data; and EK and AD drafted the work. All authors revised the work critically for important intellectual content, approved the version to be published, and agreed to be accountable for all aspects of the work in ensuring that questions related to the accuracy or integrity of any part of the work are appropriately investigated and resolved.FundingThis work was supported by the Swedish Institute, the Concern Foundation (Los Angeles), and the Cancer Research Institute (New York).
