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Conceptual model to inform Legionella-amoebae control, including the roles of extracellular vesicles in engineered water system infections

FRONTIERS IN CELLULAR AND INFECTION MICROBIOLOGY (2023)

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Summary: This study presents a new method, VFC + qPCR assay, to quantify VBNC Legionella from environmental water samples. The viability of VBNC cells was confirmed through their ATP activity and ability to infect amoeba hosts. The traditional culture and qPCR methods were unable to detect the presence of VBNC Legionella.

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Summary: Bacterial pathogens replicate within host cells by acquiring host-derived nutrients while avoiding degradation. The genus Legionella has provided fundamental insights into bacterial pathogenicity through its ability to parasitize protozoa and establish a replicative niche called the Legionella-containing vacuole (LCV). The biogenesis of LCV and intracellular replication involve rapid evasion of the endocytic pathway and acquisition of host-derived nutrients mediated by bacterial effector proteins. The co-evolution of Legionella with eukaryotic hosts has resulted in an expanded genome and a vast repertoire of effector proteins that promote LCV biogenesis, protect it from endolysosomal maturation, and mediate the acquisition of host nutrients. This minireview focuses on the mechanisms by which Legionella leverages effector proteins and hijacks host cell biology for nutrient acquisition and lysosomal degradation prevention.

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Summary: Extracellular vesicles (EVs) are essential for cell-cell communication. The roles of EVs in parasites, especially in terms of pathogenesis, are of great interest for medical insight and therapeutic intervention. However, the mechanisms of EV biogenesis in human parasitic protozoa, particularly in terms of the Endosomal Sorting Complex Required for Transport (ESCRT) machinery, remain elusive. This review focuses on the current knowledge of EV biogenesis pathways in the four main clades of human parasitic protozoa (apicomplexans, trypanosomatids, flagellates, and amoebae), highlighting the diversity in EV biogenesis and the involvement of the ESCRT system in these unique organisms.

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Summary: This study evaluated EVs produced by Acanthamoeba from different genotypes, finding that all strains released EVs regardless of genotype, with no differences in size and/or concentration detected. EVs exhibited a predominant activity of serine proteases, with higher intensity in T4 and T1 strains. EVs from the environmental, nonpathogenic T11 strain had a more proinflammatory profile compared to strains with pathogenic traits, inducing higher levels of Nitrite, tumor necrosis factor alpha, and interleukin-6 via TLR4/TLR2. Preincubation with EVs treated with protease inhibitors or heating drastically decreased nitrite concentration production in macrophages, suggesting that the immunomodulatory effects of EVs may reflect their pathogenic potential depending on the Acanthamoeba strains and are dependent on protease integrity.

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Differential Bacterial Predation by Free-Living Amoebae May Result in Blooms of Legionella in Drinking Water Systems

Mohamed Shaheen et al.

Summary: The research shows that water-related FLA species preferentially feed on Escherichia coli when the preferred bacterial food-sources are limited, although they also support the intracellular growth of Legionella pneumophila. Moreover, once Legionella pneumophila are intracellular, they inhibit the digestion of co-occurring E. coli within the same trophozoites. These findings suggest that FLA's feeding preference may lead to an increase in Legionella abundance and facilitate the internalization of Legionella into FLA within engineered water systems.

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A Methodology for Classifying Root Causes of Outbreaks of Legionnaires' Disease: Deficiencies in Environmental Control and Water Management

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Summary: A methodology was piloted to collect and interpret root causes or environmental deficiencies from Legionnaires' disease outbreak investigation reports. The study found that process failures were the majority of identified deficiencies and highlighted the importance of water management programs in preventing and mitigating outbreaks of LD. Emphasizing documentation, verification, validation, and continuous program review steps are crucial for maximizing the effectiveness of WMPs.

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