Didecyldimethylammonium Chloride- and Polyhexamethylene Guanidine-Resistant Bacteria Isolated from Fecal Sludge and Their Potential Use in Biological Products for the Detoxification of Biocide-Contaminated Wastewater Prior to Conventional Biological Treatment

Toxic shock caused by the discharge of biocide-contaminated fecal sludge (FS) from chemical toilets to conventional wastewater treatment plants (WWTP) can be a major problem in activated sludge operation. It is necessary to develop new environmental approaches to mitigate the toxicity of biocides in order to avoid degrading the performance of WWTP. “Latrina”, a chemical toilet additive containing didecyldimethylammonium chloride and polyhexamethylene guanidine, is widely used in environmentally safe toilet complexes (ESTC) on Russian railway trains to deodorize FS and control microbial activity. In this work, seven biocide-resistant bacterial strains were isolated and identified from the FS of ESTC. The values of the minimum inhibitory and bactericidal concentrations of biocides for the isolated strains were 4.5–10 times higher than for the collection microorganisms. The bacterium Alcaligenes faecalis DOS7 was found to be particularly resistant to “Latrina”, the minimum inhibitory concentration of which was almost 30 times higher than recommended for ESTC. Biological products based on isolated bacterial strains proved to be effective for FS biodegradation under both aerobic and anaerobic conditions. The results of the biochemical oxygen demand test and the newly developed disk-diffusion bioassay confirmed that isolated strains contribute to reducing toxicity of biocidal agents in FS. Hyper-resistance, non-pathogenicity, and potential plant growth-promoting ability make A. faecalis DOS7 promising for use in various biological products for wastewater treatment and bioremediation of soils contaminated with biocides, as well as in agriculture to increase plant productivity

DNA-Binding Protein Dps Protects <i>Escherichia coli</i> Cells against Multiple Stresses during Desiccation

Gradual dehydration is one of the frequent lethal yet poorly understood stresses that bacterial cells constantly face in the environment when their micro ecotopes dry out, as well as in industrial processes. Bacteria successfully survive extreme desiccation through complex rearrangements at the structural, physiological, and molecular levels, in which proteins are involved. The DNA-binding protein Dps has previously been shown to protect bacterial cells from many adverse effects. In our work, using engineered genetic models of E. coli to produce bacterial cells with overproduction of Dps protein, the protective function of Dps protein under multiple desiccation stresses was demonstrated for the first time. It was shown that the titer of viable cells after rehydration in the experimental variants with Dps protein overexpression was 1.5–8.5 times higher. Scanning electron microscopy was used to show a change in cell morphology upon rehydration. It was also proved that immobilization in the extracellular matrix, which is greater when the Dps protein is overexpressed, helps the cells survive. Transmission electron microscopy revealed disruption of the crystal structure of DNA–Dps crystals in E. coli cells that underwent desiccation stress and subsequent watering. Coarse-grained molecular dynamics simulations showed the protective function of Dps in DNA–Dps co-crystals during desiccation. The data obtained are important for improving biotechnological processes in which bacterial cells undergo desiccation