Axenic mass cultivation of the free-living soil amoeba, Acanthamoeba castellanii in a laboratory fermentor

1996 ◽  
Vol 69 (4) ◽  
pp. 317-322 ◽  
Author(s):  
Peter H. H. Weekers ◽  
John P. H. Wijen ◽  
Bart P. Lomans ◽  
Godfried D. Vogels
Keyword(s):  
Acanthamoeba Castellanii ◽  
Free Living ◽  
Mass Cultivation ◽  
Soil Amoeba ◽  
Laboratory Fermentor

scholarly journals Mitigation of Expression of Virulence Genes in Legionella pneumophila Internalized in the Free-Living Amoeba Willaertia magna C2c Maky

Pathogens ◽  
2020 ◽  
Vol 9 (6) ◽  
pp. 447 ◽  
Author(s):  
Rayane Mouh Mameri ◽  
Jacques Bodennec ◽  
Laurent Bezin ◽  
Sandrine Demanèche
Keyword(s):  
Gene Expression ◽  
Legionella Pneumophila ◽  
Virulence Genes ◽  
Natural Habitat ◽  
Acanthamoeba Castellanii ◽  
Severe Form ◽  
Aquatic Environments ◽  
Free Living ◽  
Intracellular Parasites ◽  
Free Living Amoeba

Legionella pneumophila is a human pathogen responsible for a severe form of pneumonia named Legionnaire disease. Its natural habitat is aquatic environments, being in a free state or intracellular parasites of free-living amoebae, such as Acanthamoeba castellanii. This pathogen is able to replicate within some amoebae. Willaertia magna C2c Maky, a non-pathogenic amoeba, was previously demonstrated to resist to L. pneumophila and even to be able to eliminate the L. pneumophila strains Philadelphia, Lens, and Paris. Here, we studied the induction of seven virulence genes of three L. pneumophila strains (Paris, Philadelphia, and Lens) within W. magna C2c Maky in comparison within A. castellanii and with the gene expression level of L. pneumophila strains alone used as controls. We defined a gene expression-based virulence index to compare easily and without bias the transcript levels in different conditions and demonstrated that W. magna C2c Maky did not increase the virulence of L. pneumophila strains in contrast to A. castellanii. These results confirmed the non-permissiveness of W. magna C2c Maky toward L. pneumophila strains.

scholarly journals Behavior of Yersinia enterocolitica in the Presence of the Bacterivorous Acanthamoeba castellanii

2013 ◽  
Vol 79 (20) ◽  
pp. 6407-6413 ◽  
Author(s):  
E. Lambrecht ◽  
J. Baré ◽  
I. Van Damme ◽  
W. Bert ◽  
K. Sabbe ◽  
...  
Keyword(s):  
Yersinia Enterocolitica ◽  
Pathogenic Bacteria ◽  
Acanthamoeba Castellanii ◽  
Ecological Niches ◽  
Free Living ◽  
Content Type ◽  
Food Borne ◽  
Transmission Electron ◽  
Set Up ◽  
The Impact

ABSTRACTFree-living protozoa play an important role in the ecology and epidemiology of human-pathogenic bacteria. In the present study, the interaction betweenYersinia enterocolitica, an important food-borne pathogen, and the free-living amoebaAcanthamoeba castellaniiwas studied. Several cocultivation assays were set up to assess the resistance ofY. enterocoliticatoA. castellaniipredation and the impact of environmental factors and bacterial strain-specific characteristics. Results showed that allY. enterocoliticastrains persist in association withA. castellaniifor at least 14 days, and associations withA. castellaniienhanced survival ofYersiniaunder nutrient-rich conditions at 25°C and under nutrient-poor conditions at 37°C. Amoebae cultivated in the supernatant of oneYersiniastrain showed temperature- and time-dependent permeabilization. Intraprotozoan survival ofY. enterocoliticadepended on nutrient availability and temperature, with up to 2.8 log CFU/ml bacteria displaying intracellular survival at 7°C for at least 4 days in nutrient-rich medium. Transmission electron microscopy was performed to locate theYersiniacells inside the amoebae. AsYersiniaandAcanthamoebashare similar ecological niches, this interaction identifies a role of free-living protozoa in the ecology and epidemiology ofY. enterocolitica.

scholarly journals Cellular Response of the Amoeba Acanthamoeba castellanii to Chlorine, Chlorine Dioxide, and Monochloramine Treatments

2011 ◽  
Vol 77 (14) ◽  
pp. 4974-4980 ◽  
Author(s):  
Emerancienne Mogoa ◽  
Charles Bodet ◽  
Franck Morel ◽  
Marie-Hélène Rodier ◽  
Bernard Legube ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Flow Cytometry ◽  
Chlorine Dioxide ◽  
Mode Of Action ◽  
Cellular Response ◽  
Acanthamoeba Castellanii ◽  
Size Reduction ◽  
Resistant Bacteria ◽  
Free Living ◽  
Content Type

ABSTRACTAcanthamoeba castellaniiis a free-living amoebae commonly found in water systems. Free-living amoebae might be pathogenic but are also known to bear phagocytosis-resistant bacteria, protecting these bacteria from water treatments. The mode of action of these treatments is poorly understood, particularly on amoebae. It is important to examine the action of these treatments on amoebae in order to improve them. The cellular response to chlorine, chlorine dioxide, and monochloramine was tested onA. castellaniitrophozoites. Doses of disinfectants leading to up to a 3-log reduction were compared by flow cytometry and electron microscopy. Chlorine treatment led to size reduction, permeabilization, and retraction of pseudopods. In addition, treatment with chlorine dioxide led to a vacuolization of the cytoplasm. Monochloramine had a dose-dependent effect. At the highest doses monochloramine treatment resulted in almost no changes in cell size and permeability, as shown by flow cytometry, but the cell surface became smooth and dense, as seen by electron microscopy. We show that these disinfectants globally induced size reduction, membrane permeabilization, and morphological modifications but that they have a different mode of action onA. castellanii.

scholarly journals Isoniazid Conjugated Magnetic Nanoparticles Loaded with Amphotericin B as a Potent Antiamoebic Agent against Acanthamoeba castellanii

Antibiotics ◽  
2020 ◽  
Vol 9 (5) ◽  
pp. 276
Author(s):  
Kawish Iqbal ◽  
Sumayah Abdelnasir Osman Abdalla ◽  
Ayaz Anwar ◽  
Kanwal Muhammad Iqbal ◽  
Muhammad Raza Shah ◽  
...  
Keyword(s):  
Magnetic Nanoparticles ◽  
Amphotericin B ◽  
Central Nervous System Infection ◽  
Acanthamoeba Castellanii ◽  
Acanthamoeba Keratitis ◽  
Free Living ◽  
Granulomatous Amoebic Encephalitis ◽  
Nano Drug Delivery ◽  
Free Living Amoeba ◽  
Acanthamoeba Species

The pathogenic free-living amoeba, Acanthamoeba castellanii, is responsible for a rare but deadly central nervous system infection, granulomatous amoebic encephalitis and a blinding eye disease called Acanthamoeba keratitis. Currently, a combination of biguanides, amidine, azoles and antibiotics are used to manage these infections; however, the host cell cytotoxicity of these drugs remains a challenge. Furthermore, Acanthamoeba species are capable of transforming to the cyst form to resist chemotherapy. Herein, we have developed a nano drug delivery system based on iron oxide nanoparticles conjugated with isoniazid, which were further loaded with amphotericin B (ISO-NPs-AMP) to cause potent antiamoebic effects against Acanthamoeba castellanii. The IC50 of isoniazid conjugated with magnetic nanoparticles and loaded with amphotericin B was found to be 45 μg/mL against Acanthamoeba castellanii trophozoites and 50 μg/mL against cysts. The results obtained in this study have promising implications in drug discovery as these nanomaterials exhibited high trophicidal and cysticidal effects, as well as limited cytotoxicity against rat and human cells.

scholarly journals Encystment Induces Down-Regulation of an Acetyltransferase-Like Gene in Acanthamoeba castellanii

Pathogens ◽  
2020 ◽  
Vol 9 (5) ◽  
pp. 321 ◽  
Author(s):  
Steven Rolland ◽  
Luce Mengue ◽  
Cyril Noël ◽  
Stéphanie Crapart ◽  
Anne Mercier ◽  
...  
Keyword(s):  
Bioinformatics Analysis ◽  
Acanthamoeba Castellanii ◽  
Acanthamoeba Keratitis ◽  
Free Living ◽  
Adverse Conditions ◽  
Genetic Modifications ◽  
Granulomatous Amoebic Encephalitis ◽  
Free Living Amoeba ◽  
Causative Agents

Acanthamoeba castellanii is a ubiquitous free-living amoeba. Pathogenic strains are causative agents of Acanthamoeba keratitis and granulomatous amoebic encephalitis. In response to adverse conditions, A. castellanii differentiate into cysts, which are metabolically inactive and resistant cells. This process, also named encystment, involves biochemical and genetic modifications that remain largely unknown. This study characterizes the role of the ACA1_384820 Acanthamoeba gene during encystment. This gene encodes a putative N-acetyltransferase, belonging to the Gcn5-related N-acetyltransferase (GNAT) family. We showed that expression of the ACA1_384820 gene was down-regulated as early as two hours after induction of encystment in A. castellanii. Interestingly, overexpression of the ACA1_384820 gene affects formation of cysts. Unexpectedly, the search of homologs of ACA1_384820 in the Eukaryota gene datasets failed, except for some species in the Acanthamoeba genus. Bioinformatics analysis suggested a possible lateral acquisition of this gene from prokaryotic cells. This study enabled us to describe a new Acanthamoeba gene that is down-regulated during encystment.

scholarly journals Oxygen affinity of the respiratory chain of Acanthamoeba castellanii

1983 ◽  
Vol 214 (1) ◽  
pp. 47-51 ◽  
Author(s):  
D Lloyd ◽  
H Mellor ◽  
J L Williams
Keyword(s):  
Plasma Membrane ◽  
Cytochrome C ◽  
Cytochrome B ◽  
Respiratory Chain ◽  
Oxygen Affinity ◽  
Acanthamoeba Castellanii ◽  
Intact Cells ◽  
Cytochrome Aa3 ◽  
Isolated Mitochondria ◽  
Soil Amoeba

Apparent Km values for O2 for the soil amoeba Acanthamoeba castellanii determined polarographically and by bioluminescence gave similar values (0.37 and 0.41 microM respectively). Mitochondria oxidizing succinate or NADH in the presence or absence of ADP gave values in the range 0.21-0.36 microM-O2. Oxidation of respiratory-chain components to 50% of the aerobic steady states in intact cells was observed at the following O2 concentrations: cytochrome aa3, 0.1-0.25 microM; cytochrome c, 0.3-0.6 microM; cytochrome b, 0.35-0.45 microM; flavoprotein, 2 microM. In isolated mitochondria corresponding values for a-, c- and b-type cytochromes were 0.007, 0.035-0.05 and 0.06-0.09 microM-O2. It is concluded that an O2 gradient exists between plasma membrane and mitochondria in A. castellanii.

scholarly journals Legionella pneumophila Contains a Type II General Secretion Pathway Required for Growth in Amoebae as Well as for Secretion of the Msp Protease

1999 ◽  
Vol 67 (7) ◽  
pp. 3662-3666 ◽  
Author(s):  
Laura M. Hales ◽  
Howard A. Shuman
Keyword(s):  
Legionella Pneumophila ◽  
Acanthamoeba Castellanii ◽  
Gram Negative Bacteria ◽  
Type Ii ◽  
Free Living ◽  
Gram Negative ◽  
Secretion Pathway ◽  
Free Living Amoeba ◽  
Substitution Mutation

ABSTRACT We report the identification of a set of Legionella pneumophila genes that encode products with homology to proteins of the type II general secretion pathway of gram-negative bacteria. A strain containing a deletion-substitution mutation of two of these genes was unable to secrete the Msp protease. This strain was unable to multiply within the free-living amoeba Acanthamoeba castellanii yet was able to kill HL-60-derived macrophages. Because Msp is not required for growth in amoebae, other proteins which are important for growth in amoebae are likely secreted by this pathway.

scholarly journals Phagocyte chase behaviours: discrimination between Gram-negative and Gram-positive bacteria by amoebae

2019 ◽  
Vol 15 (1) ◽  
pp. 20180607 ◽  
Author(s):  
Ghazal Rashidi ◽  
Elizabeth A. Ostrowski
Keyword(s):  
Prior Work ◽  
Free Living ◽  
Gram Positive ◽  
Gram Negative ◽  
Gram Positive Bacteria ◽  
Different Types ◽  
Soil Amoeba ◽  
Response Profiles ◽  
Mammalian Immune System ◽  
Geographical Locations

Phagocytes are cells that pursue, engulf and kill bacteria. They include macrophages and neutrophils of the mammalian immune system, as well as free-living amoebae that hunt and engulf bacteria for food. Phagocytosis can result in diverse outcomes, ranging from sustenance to infection and colonization by either pathogens or beneficial symbionts—and thus, discrimination may be necessary to seek out good bacteria while avoiding bad ones. Here we tested whether the soil amoeba Dictyostelium discoideum can discriminate among different types of bacteria using behavioural assays where amoebae were presented with paired choices of different bacteria. We observed variation in the extent to which the amoebae pursued different types of bacteria, as well as preferential migration towards Gram-negative compared with Gram-positive bacteria. Response profiles were similar for amoebae that originated from different geographical locations, suggesting that chase preference is conserved across much of the species range. While prior work has demonstrated that bacteria use chemotaxis to seek out amoebae they colonize, our work suggests that the opposite also occurs—amoebae can preferentially direct themselves to particular bacteria in the environment. Preferential sensing and response may help to explain why some amoeba–bacteria associations are more common in nature than others.

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