In this study, we investigated biodeterioration of materials used in tempera painting by analyzing the structure of the microbiome in ancient tempera paintings exhibited in State Tretyakov Gallery, Moscow, Russia. Samples were obtained from 16 th-century paintings, including a grand Russian Orthodox icon "The Church Militant" (all exhibits were without visible signs of biodeterioration), and from surrounding walls and ceilings (with vast zones of visible microbial growth). A number of microorganisms isolated from visible signs of environmental bio-damage were also detected in tempera paintings kept in temperature-and humidity-controlled conditions unfavorable for the growth of microflora. To determine the biodegrading potential of the microbiome for tempera paintings, we developed a set of mock layers from paintwork materials used in tempera painting of 16 th century and their modern analogues and inoculated them with cultures containing filamentous fungi and bacteria. The susceptibility to microbial degradation of individual tempera painting materials was examined by micro-Fourier Transform Infrared (FTIR) spectroscopy, which enabled detection of even invisible signs of biodeterioration. The results indicate that the microorganisms isolated from paintings and surrounding areas in the museum are capable of causing significant damage of various tempera materials, among which varnishes were the most resistant; however, the addition of antiseptic (sodium pentachlorophenolate) can inhibit microbial growth on sturgeon glue.

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