Characterization of the natural bacterial microbiota of pathogenic free-living amoebae (Acanthamoeba spp. and Naegleria fowleri) isolated from rivers and tap water in Guadeloupe

The Science of The Total Environment, Journal Year: 2025, Volume and Issue: 975, P. 179204 - 179204

Published: April 7, 2025

Free-living amoebae (FLA) are ubiquitous protists found in water, feeding mainly on bacteria. While most FLA harmless, Acanthamoeba spp. and Naegleria fowleri can cause keratitis and/or meningitis. host amoeba-resistant bacteria (ARB), but their natural bacterial microbiota is largely unknown. This study aimed to identify the of fowleri, castellanii, lenticulata, sp. T17, isolated from untreated (rivers) treated (tap) waters Guadeloupe. The whole water source grown with E. coli under axenic culture conditions, during successive passages, were characterized using 16S rRNA gene metabarcoding. culturable subset ARB was identified by mass spectrometry (MALDI-TOF MS) followed conventional PCR, antibiotic resistance analyzed disk diffusion method. Transmission electron microscopy used locate within amoebae. metabarcoding analyses Salmonella, Enterobacter Klebsiella genera as abundant waters. However, frequently detected (ARB) Bosea, Escherichia-Shigella, Microbacterium, Pseudomonas genera. Our findings revealed, for first time, occurrence several N. including Escherichia coli. Additionally, we Legionella A. castellanii Bordetella lenticulata. four pathogenic showed both temporary permanent associations various genera, depending number passages conditions. species distinct exhibited different antibiotics. cytoplasm trophozoites. presence Guadeloupe's drinking systems pose health risks. results highlight need regular monitoring ensure safety understanding amoebae-bacteria interactions better management. also questions immune response amoeba infection.

Language: Английский

Recombinant protein expression in Acanthamoeba castellanii

Frontiers in Bioengineering and Biotechnology, Journal Year: 2025, Volume and Issue: 13

Published: March 20, 2025

The ongoing quest to improve protein production efficiency, quality, and versatility fuels the exploration of novel expression systems. In this research, we explored potential axenically culturable Acanthamoeba as an alternative for producing recombinant eukaryotic proteins. We constructed plasmid vectors utilizing TBP promoter facilitate within protozoan system. Our primary objectives were develop efficient transfection method assess capacity castellanii glycoprotein expression. initial efforts yielded successful firefly luciferase reporter gene, allowing us optimize protocol. Subsequently, compared Chikungunya virus E2 across three systems: E. coli , Acanthamoeba, mammalian cells. Interestingly, expressed in exhibited a molecular weight higher than bacterial cells but lower cells, suggesting possibility glycosylation occurring These findings collectively suggest that protozoa, like A. represent promising avenue developing low-cost

Language: Английский