Proteomics and Lipidomics Investigations to Decipher the Behavior of Willaertia magna C2c Maky According to Different Culture Modes.

Issam Hasni,Amina Cherif Louazani,Philippe Decloquement,Olivier Abbe,Bernard La Scola,Sandrine Demanèche,Nicholas Armstrong,Said Azza,Philippe Colson,Anthony Fontanini,Eric Chabrière

doi:10.3390/microorganisms8111791

Abstract

Willaertia magna C2c Maky is a free-living amoeba that has demonstrated its ability to inhibit the intracellular multiplication of some Legionella pneumophila strains, which are pathogenic bacteria inhabiting the aquatic environment. The Amoeba, an industry involved in the treatment of microbiological risk in the water and plant protection sectors, has developed a natural biocide based on the property of W. magna to manage the proliferation of the pathogen in cooling towers. In axenic liquid medium, amoebas are usually cultivated in adhesion on culture flask. However, we implemented a liquid culture in suspension using bioreactors in order to produce large quantities of W. magna. In order to investigate the culture condition effects on W. magna, we conducted a study based on microscopic, proteomics and lipidomics analyzes. According to the culture condition, amoeba exhibited two different phenotypes. The differential proteomics study showed that amoebas seemed to promote the lipid metabolism pathway in suspension culture, whereas we observed an upregulation of the carbohydrate pathway in adherent culture. Furthermore, we observed an over-regulation of proteins related to the cytoskeleton for W. magna cells grown in adhesion. Regarding the lipid analysis, suspension and adhesion cell growth showed comparable lipid class compositions. However, the differential lipid analysis revealed differences that confirmed cell phenotype differences observed by microscopy and predicted by proteomics. Overall, this study provides us with a better insight into the biology and molecular processes of W. magna in different culture lifestyles.

Highlights

Willaertia magna is a free-living amoeba (FLA) belonging to the class Heterolobosea and family Vahlkampfiidae [1,2]
W. magna is phylogenetically close to Naegleria fowleri, a virulent amoeba causing primary amoebic meningoencephalitis (PAM) in humans, in vivo and in silico experiments have demonstrated the non-pathogenicity of W. magna [4,7,8]
The length of the amoebas cultivated under two different conditions exhibited approximately similar size, Micwroiotrhganliesnmgs t2h020o,f8,21179μ1 m (+/−3.34) and 19.5 μm (+/−4.50) for the amoeba grown in adhesion a6nodf 16 suspension, respectively (Table S4). sizaem, woeiHtbhoaswleiennvgbetirho, rtoehafec2at1omrμo(m5eb.2a(μ+inm/−a)3.d.Wh34ees)ioaobnnsde(1r1v49.e2.d5μtμmhme) pis(r+eas/p−epn4rc.o5ex0oi)mffpaostreeltuyhde3oatpimomdoeiseabwfaoirdgatrhmowtohenabnaintshiaendcahudelhtsieivosaintoenad,nd suswpheincshioisnc, hreasrapcetcetriivsetilcyo(fTtahbeleacSt4iv).e movement of the amoebas

Summary

Introduction

Willaertia magna is a free-living amoeba (FLA) belonging to the class Heterolobosea and family Vahlkampfiidae [1,2]. This FLA is found in natural and artificial environments, such as humid soil, bovine feces, composts, thermal waters, and fresh water sediments [3,4,5]. The trophozoites stage is characterized by a large form containing one or several nuclei, food and contractile vacuoles, and mitochondria. The amoeboid form can temporary transform into ameboflagellate under certain conditions [6]. The trophozoite has the capacity to transform into large cysts containing pores in the cell wall [4]. The analysis of the W. magna draft genome revealed a DNA length of 36.5 megabases and less than 19,000 genes [8]

Methods

Results

Conclusion