Search

Extracellular vesicles (EVs or exosomes) are well described for bacterial pathogens associated with our gastrointestinal system, and more recently as a novel mechanism for environmental persistence, dissemination and infection for human enteric viruses. However, the roles played by EVs in the ancient arms race that continues between amoebae and one of their prey, Legionella pneumophila, is poorly understood. At best we know of intracellular vesicles of amoebae containing a mix of bacterial prey species, which also provides an enhanced niche for bacteriophage infection/spread. Free-living amoeba-associated pathogens have recently been recognized to have enhanced resistance to disinfection and environmental stressors, adding to previously understood (but for relatively few species of) bacteria sequestered within amoebal cysts. However, the focus of the current work is to review the likely impacts of large numbers of respiratory-sized EVs containing numerous L. pneumophila cells studied in pure and biofilm systems with mixed prey species. These encapsulated pathogens are orders of magnitude more resistant to disinfection than free cells, and our engineered systems with residual disinfectants could promote evolution of resistance (including AMR), enhanced virulence and EV release. All these are key features for evolution within a dead-end human pathogen post lung infection. Traditional single-hit pathogen infection models used to estimate the probability of infection/disease and critical environmental concentrations via quantitative microbial risk assessments may also need to change. In short, recognizing that EV-packaged cells are highly virulent units for transmission of legionellae, which may also modulate/avoid human host immune responses. Key data gaps are raised and a previous conceptual model expanded upon to clarify where biofilm EVs could play a role promoting risk as well as inform a more wholistic management program to proactively control legionellosis.

• CT kinetics of SO 4 •− and •OH-driven disinfection of fungi and bacteria were established and modeled. • •OH played a more significant role in the PMS/Me(II) disinfection systems than SO 4 •−. • The differences in disinfection resistance between bacteria and fungal spores were compared. • Antioxidant mechanisms and external structures made fungal spores more resistant to radicals attack. Peroxymonosulfate(PMS)-based advanced oxidation process have been recognized as efficient disinfection processes. This study comprehensively investigated the role of sulfate radical (SO 4 •−) and hydroxyl radical (•OH)-driven disinfection of bacteria and fungal spores by the PMS/metals ions (Me(II)) systems and modeled the CT value based on the relationship between survival and ∫ [ Radical ] dt, with the aim to provide an accurate and quantitative kinetic data of inactivation processes. The results indicated that •OH played a more central role than SO 4 •− in the inactivation process, and bacteria were more vulnerable to radical attack than fungal spores due to the differences in antioxidant mechanisms and external structures. The k value of •OH -induced inactivation of E. coli was approximately 3-fold higher than that of A. niger , and the shoulder length of •OH -induced inactivation of E. coli was closely 52-fold shorter than that of A. niger after treated with the PMS/Co(II) system. The morphological and biochemical changes revealed that PMS/Me(II) treatment caused membrane damage, intracellular ROS accumulation and esterase activity loss in microorganisms. This study significantly improved the understanding of the contribution of radicals in the process of microbial inactivation by PMS/Me(II) and would provide important implications for the further development of technologies to cope with the highly resistant fungal spores in drinking water. [Display omitted] [ABSTRACT FROM AUTHOR]

Extracellular vesicles (EVs or exosomes) are well described for bacterial pathogens associated with our gastrointestinal system, and more recently as a novel mechanism for environmental persistence, dissemination and infection for human enteric viruses. However, the roles played by EVs in the ancient arms race that continues between amoebae and one of their prey, Legionella pneumophila , is poorly understood. At best we know of intracellular vesicles of amoebae containing a mix of bacterial prey species, which also provides an enhanced niche for bacteriophage infection/spread. Free-living amoeba-associated pathogens have recently been recognized to have enhanced resistance to disinfection and environmental stressors, adding to previously understood (but for relatively few species of) bacteria sequestered within amoebal cysts. However, the focus of the current work is to review the likely impacts of large numbers of respiratory-sized EVs containing numerous L. pneumophila cells studied in pure and biofilm systems with mixed prey species. These encapsulated pathogens are orders of magnitude more resistant to disinfection than free cells, and our engineered systems with residual disinfectants could promote evolution of resistance (including AMR), enhanced virulence and EV release. All these are key features for evolution within a dead-end human pathogen post lung infection. Traditional single-hit pathogen infection models used to estimate the probability of infection/disease and critical environmental concentrations via quantitative microbial risk assessments may also need to change. In short, recognizing that EV-packaged cells are highly virulent units for transmission of legionellae, which may also modulate/avoid human host immune responses. Key data gaps are raised and a previous conceptual model expanded upon to clarify where biofilm EVs could play a role promoting risk as well as inform a more wholistic management program to proactively control legionellosis., Competing Interests: The author declares that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Ashbolt.)

In this paper, for the first time, we show in chloraminated systems, the chloramine decaying proteins (CDP) play an important role in bulk water and biomass (biofilm) in resisting disinfectant. Extracellular polymeric substances in biofilm/biomass are known to protect microbes from disinfectants and toxic materials, but the exact mechanism(s) is/are not known. Starting with the seed from a nitrifying chloraminated reactor, two 5 L reactors were fed intermittently with either chloramine or ammonia containing nutrient solution. The degree of nitrification increased with time in both reactors despite an increase in soluble CDP in the chloraminated reactor, while soluble CDP decreased in the ammoniated one. The suspended biomass collected after eight months of operation from chloraminated reactor contained CDP and responded to short-term chloramine stress (1.5 h with initial 1.5 mg-Cl 2 ·L−1) by the additional production of soluble CDP. The suspended biomass from ammoniated reactor neither contained CDP nor produced soluble CDP as a stress response. The production, release and accumulation of CDP in biomass (biofilm) could be one of several mechanisms microbes use to defend against disinfectants (stress). The new understanding will pave the way for better disinfection management and better design of experiments. Image 1 • Chloramine decaying Protein (CDP) existed in chloraminated system. • Chloramine dosing in chloraminated reactor induced the production of CDP. • Both bulk water and suspended biomass from chloraminated reactor contained CDP. • Short term chloramine stress further induced CDP production in biomass. • In the absence of chloramine, in ammoniated reactor, CDP gradually disappeared. [ABSTRACT FROM AUTHOR]