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Lisina T.O., Gladkov G.V., Orlova O.V., Kimeklis A.K., Kichko A.A., Andronov E.E. Effects of preservation and storage methods on the microbiome functionality and composition of BAGS, a bioactive formulation. Sel'skokhozyaistvennaya Biologiya [Agricultural Biology], 2026, Vol. 61, № 3, p. 521-533.
(how to cite this article)

doi: 10.15389/agrobiology.2026.3.521eng

UDC: 579.64:631.461.61:57.083.1

Acknowledgements:
The research was conducted using equipment from the “Genomic Technologies, Proteomics, and Cell Biology” Research Center at the Federal State Budgetary Scientific Institution All-Russian Research Institute of Agricultural Microbiology.
Funded by grant 23-16-00147 from the Russian Science Foundation.

 

EFFECTS OF PRESERVATION AND STORAGE METHODS ON THE MICROBIOME FUNCTIONALITY AND COMPOSITION OF BAGS, A BIOACTIVE FORMULATION

T.O. Lisina ✉, G.V. Gladkov, O.V. Orlova, A.K. Kimeklis,
A.A. Kichko, E.E. Andronov

All-Russian Research Institute for Agricultural Microbiology, 3, sh. Podbel’skogo, St. Petersburg, 196608 Russia, e-mail lisina-to@yandex.ru (✉ corresponding author) grgladkov@arriam.ru,
falenki@hotmail.com, kimeklis@gmail.com, 2014arki@gmail.com, eeandr@gmail.com, eeandr@gmail.com

ORCID:
Lisina T.O. orcid.org/0000-0003-1268-4166
Kimeklis A.K. orcid.org/0000-0003-0348-7021
Gladkov G.V. orcid.org/0000-0002-5489-1414
Kichko A.A. orcid.org/0000-0002-8482-6226
Orlova O.V. orcid.org/0000-0002-2154-503Х
Andronov E.E. orcid.org/0000-0002-5204-262Х

Final revision received October 21, 2025
Accepted November 15, 2025

Currently, methods for the preservation and storage of microorganisms have been developed and optimized, based on inducing a state of anabiosis in cells, during which vital processes are partially (storage at low temperatures) or completely (drying, cryopreservation) suspended. However, only a few studies have addressed the preservation of complex microbial consortia while maintaining the properties that ensure their effectiveness. The ability to analyze the taxonomic composition of complex microbial communities became possible with the development of high-throughput sequencing techniques/. In this study, data on the possibility of conserving stock samples of the BAGS were obtained for the first time. The study aimed to test different methods of storage and preservation of the BAGS fertilizer and to estimate their effect on cellulolytic activity and the taxonomic composition of the prokaryotic component of the consortium using deep sequencing of 16S rRNA gene amplicon libraries. The study was conducted in 2024-2025 at the All-Russian Research Institute of Agricultural Microbiology. The objects of the study were: the initial mature microbial BAGS (first generation), obtained as a result of composting a peat-straw-mineral mixture for 6 months and inoculum (previously prepared BAGS); reactivated BAGS samples after storage by different methods for 6 months; experimental batches of BAGS obtained by composting a peat-straw-mineral mixture using samples of the fertilizer  fertilizer reactivated after preservation as inoculum. Three methods of storage and preservation of the initial fertilizer BAGS samples were tested: storage of the fresh fertilizer fertilizer in a refrigerator at +4 °C; drying at room temperature to an at room temperature for 1 week, during which the pre-dried samples were moistened with water to 60 % of their total water-holding capacity. The reactivated samples were used to inoculate a peat-straw-mineral mixture during the preparation of experimental BAGS batches. To obtain a control batch of BAGS (3rd generation), a fresh fertilizer h of the 2nd generation was used as inoculum; it was prepared using the standard method simultaneously with the storage of the experimental samples. All BAGS batches were incubated at 28 °C for 5 months. Cellulolytic activity was assessed monthly using two methods (the modified Christensen method and the application method). Microbiological analysis of the finished 5-month preparations was performed using deep sequencing of 16S rRNA gene amplicon libraries. Quantitative comparison of the microbiomes was carried out based on alpha- (within-sample) and beta-diversity (between-sample) indices. To calculate alpha-diversity indices, the amplicon libraries were normalized using the minimum size rarefaction method. The assessment of alpha-diversity included the determination of the total number of ASVs and the Simpson’s inverse index. Beta-diversity was visualized using non-metric multidimensional scaling (NMDS) based on the Bray-Curtis metric. An effective BAGS batch with high cellulolytic activity was obtained by using a reactivated sample after drying as the inoculum. The species diversity of the communities in all three experimental BAGS batches was twofold higher than that of the control. Similarity in taxon was observed across all variants at the phylum level. However, based on the beta-diversity indices, all communities composition differed in their microbial composition. The most comparable were the consortia of the control preparation and the BAGS fertilizer made using inoculum after drying. Representatives of the microbiome—markers of an effective fertilizer associated with nitrogen metabolism and the breakdown of complex polysaccharides—were identified: MND1, Methylomirabilota, Cryseolinea, Gracillibacteria.

Keywords: BAGS preparation, drying, cryopreservation, cellulolytic activity, taxonomic composition, species diversity.

 

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