Development and Substantiation of the Methodology for Automated Probiotic-Based Cleaning of Indoor Environments

Introduction: Cleaning of the indoor environment with probiotic agents is an evolving environmentally friendly disinfection technology aimed at improving epidemiological and hygienic conditions by displacing pathogenic microflora from environmental objects with probiotic cultures.
Objective: To develop and validate the methodology for an automated probiotic cleaning of indoor environment using Bacillus spp. designed for a modern network software and hardware complex based on the Internet of Things system and to test it under simulating real-life application conditions on laboratory animals.
Materials and Methods: Having reviewed 32 literature sources, we estimated the regimen for automated probioticbased purification and tested the new technique in a 40 m3 closed experimental chamber. The solution of a commercial probiotic of Bacillus spp. was sprayed using an adiabatic humidifier connected to the control module of the Internet of Things system. We established the antimicrobial efficacy of cleaning using standard microbiological testing methods and evaluated health effects using integral indicators of the functional state and changes in gut microbiota in laboratory rats.
Results: We developed and substantiated the methodology for automated probiotic cleaning of indoor environment using Bacillus spp. To clean a 40 m3 room, 273 mL of the Bacillus-based probiotic cleanser containing (2.8 ± 0.4) × 10³ CFU/mL of microorganisms should be sprayed hourly for 28 days. Testing of this automated mode proved the reduction in the surface bioburden of Enterococcus spp. and Staphylococcus spp. with no excessive growth of Bacillus spp. No adverse health effects were observed in laboratory animals. The probiotic demonstrated anticlostridial activity in the large intestine of rats.
Conclusions: The developed and validated new methodology of automated probiotic-based cleaning of the indoor environment has undergone real-life scenario testing. Basic data on its impact on indoor pathogens and some physiological parameters of laboratory animals were collected.

1. Shashina TA, Novikov SM, Matsyuk AV, Lando NG. Guidelines for the assessment of regional factors of the urban population exposure. Gigiena i Sanitariya. 2007;(5):20-24. (In Russ.)

2. Lee S, Lee K. Seasonal differences in determinants of time location patterns in an urban population: A large population-based study in Korea. Int J Environ Res Public Health. 2017;14(7):672. doi: 10.3390/ijerph14070672

3. Raju S, Siddharthan T, McCormack MC. Indoor air pollution and respiratory health. Clin Chest Med. 2020;41(4):825-843. doi: 10.1016/j.ccm.2020.08.014

4. Kumar P, Singh AB, Singh R. Comprehensive health risk assessment of microbial indoor air quality in microenvironments. PLoS One. 2022;17(2):e0264226. doi: 10.1371/journal.pone.0264226

5. Rathbone CJ, Bousiotis D, Rose OG, Pope FD. Using lowcost sensors to assess common air pollution sources across multiple residences. Sci Rep. 2025;15(1):1803. doi: 10.1038/s41598-025-85985-1

6. Caselli E. Hygiene: Microbial strategies to reduce pathogens and drug resistance in clinical settings. Microb Biotechnol. 2017;10(5):1079-1083. doi: 10.1111/1751-7915.12755

7. Falagas ME, Makris GC. Probiotic bacteria and biosurfactants for nosocomial infection control: A hypothesis. J Hosp Infect. 2009;71(4):301-306. doi: 10.1016/j.jhin.2008.12.008

8. Ilyakova AV, Shestopalov NV, Fedorova LS, Belova AS. The possibility of using bacteria Bacillus in the production of disinfectants. Gigiena i Sanitariya. 2020;99(5):436-442. (In Russ.) doi: 10.33029/0016-9900-2020-99-5-436-442

9. Torres-Sánchez A, Pardo-Cacho J, López-Moreno A, Ruiz-Moreno A, Cerk K, Aguilera M. Antimicrobial effects of potential probiotics of Bacillus spp. isolated from human microbiota: In vitro and in silico methods. Microorganisms. 2021;9(8):1615. doi: 10.3390/microorganisms9081615

10. Soffritti I, D’Accolti M, Cason C, et al. Introduction of probiotic–based sanitation in the emergency ward of a children’s hospital during the COVID–19 pandemic. Infect Drug Resist. 2022;15:1399-1410. doi: 10.2147/idr.s356740

11. Leistner R, Kohlmorgen B, Brodzinski A, et al. Environmental cleaning to prevent hospital-acquired infections on non-intensive care units: A pragmatic, single-centre, cluster randomized controlled, crossover trial comparing soap-based, disinfection and probiotic cleaning. EClinicalMedicine. 2023;59:101958. doi: 10.1016/j.eclinm.2023.101958

12. Cowell N, Chapman L, Bloss W, Srivastava D, Bartingtonb S, Singh A. Particulate matter in a lockdown home: Evaluation, calibration, results and health risk from an IoT enabled lowcost sensor network for residential air quality monitoring. Environ Sci Atmos. 2023;3:65-84. doi: 10.1039/d2ea00124a

13. Bobulski J, Szymoniak S, Pasternak K. An IoT system for air pollution monitoring with safe data transmission. Sensors (Basel). 2024;24(2):445. doi: 10.3390/s24020445

14. Zhukova ES, Shcherbatyuk TG, Pozdniakova MA. The relationship between a malignant tumor growth and peculiarities of behavior of laboratory animals. Prirodnye Resursy Zemli i Okhrana Okruzhayushchey Sredy. 2021;2(1):44-47. (In Russ.) doi: 10.26787/nydha-2713-203x-2021-2-1-44-47

15. Redina OE, Smolenskaya SE, Markel AL. Genetic control of the behavior of ISIAH rats in the open field test. Russian Journal of Genetics. 2022;58(7):791-803. doi: 10.1134/s1022795422070146

16. Tochilina AG, Belova IV, Solovieva IV, Zhirnov VA, Ivanova TP. [Criteria for assessing the composition of the biocenosis of the lumen of the large intestine.] Spravochnik Zaveduyushchego KDL. 2016;(8):54-78. (In Russ.)

17. Kim AD, Goldberg OA, Lepekhova SA, et al. Peculiarities of topographic anatomy and crypt compartment of distal colon in Wistar rats. Byulleten’ Vostochno-Sibirskogo Na­ uchnogo Tsentra Sibirskogo Otdeleniya Rossiyskoy Akademii Meditsinskikh Nauk. 2016;1(2(108)):48-54. (In Russ.)

18. Makarova MN, Kryshen KL, Alyakrinskaya AA, Rybakova AV, Makarov VG. Characteristics of the intestinal microflora in humans and laboratory animals. Mezhdunarodnyy Vestnik Veterinarii. 2016;(4):97-105. (In Russ.)

19. Zavgorodnyaya EF. [Intestinal dysbiosis (review).] Dal’nevostochnyy Zhurnal Infektsionnoy Patologii. 2010;(16(16)):131-141. (In Russ.)

20. Zabokritskiy NA. The biologically active substances produced probiotic microorganisms of the genera Bacillus and Lactobacillus. Zhurnal Nauchnykh Statey Zdorov’e i Obrazovanie v XXI Veke. 2015;17(3):80-90. (In Russ.)

21. Zhu J, Chen Y, Imre K, et al. Mechanisms of probiotic Bacillus against enteric bacterial infections. One Health Adv. 2023;1:21. doi: 10.1186/s44280-023-00020-0

22. Kalinina NV, Zagainova AV, Yudin SM, Gaponova EB, Bunin IM. Hygienic assessment of mycological contamination of the internal environment of residential and public buildings. Gigiena i Sanitariya. 2023;102(7):632-638. (In Russ.) doi: 10.47470/0016-9900-2023-102-7-632-638

23. Francois M, Canal Delgado I, Shargorodsky N, Leu CS, Zeltser L. Assessing the effects of stress on feeding behaviors in laboratory mice. eLife. 2022;11:e70271. doi: 10.7554/eLife.70271

24. Koterov AN, Ushenkova LN, Zubenkova ES, Wainson AA, Biryukov AP, Samoylov AS. Dependence of body weight on age for random-bred albino rat and for eight lines of laboratory rat: Synthetic studies of data from experimental works and nurseries in aspect of the relationship with radiosensitivity. Some characteristics of rat species. Meditsinskaya Radiologiya i Radiatsionnaya Bezopasnost’. 2018;63(2):1-41. (In Russ.) doi: 10.12737/article_5ac6190e95da25.42157674

25. Umriukhin PE, Grigorchuk OS. Open field behavior as a predictive criteria reflecting rat corticosteronelevel before and after stress. Rossiyskiy Fiziologicheskiy Zhurnal im. I.M. Sechenova. 2015;101(12):1366-1371. (In Russ.)

26. Yildyrym EA, Laptev GYu, Novikova NI, et al. Protecting poultry against clostridiosis. Ptitsevodstvo. 2021;(12):35-38. (In Russ.)