Abstract
The discovery of several giant amoeba viruses has opened up a novel area in the field of virology. Despite this, knowledge about ecology of these viruses remains patchy. In this study, we aimed to characterize the diversity of giant viruses in Algeria by inoculating 64 environmental samples on various amoeba strains. After isolation by co-culture with nine amoeba supports, flow cytometry and electron microscopy were used to putatively identify viruses. Definitive identification was performed by PCR and sequencing. Mimiviruses, marseilleviruses, faustoviruses and cedratviruses were the main viruses isolated in this study. Moreover, a new virus, which we named fadolivirus, was also isolated and was found to belong to the recent metagenomic descriptions of Klosneuvirinae. Despite the use of 9 amoeba supports for co-culture, most of the isolates were obtained from two amoebas: Acanthamoeba castellanii Neff and Vermamoeba vermiformis CDC 19. Finally, the viruses most frequently isolated were marseilleviruses (55.5%) and Mimiviruses (22.2%). This work shows that the isolation of viruses previously detected by metagenomic analyses can be tedious, but possible.
Highlights
Megavirales order has recently been proposed to replace the current nucleocytoplasmic large DNA viruses (NCLDV) based on capsid and genome sizes, ancestral genes encoding autonomy transcription and even translation components [1,2,3,4,5,6]
Viruses (NCLDV) based on capsid and genome sizes, ancestral genes encoding autonomy transcription and even translation components [1,2,3,4,5,6]. This order contains viruses considered giant viruses and the story began at the beginning of the 21st century with the discovery of Acanthamoeba polyphaga mimivirus (APMV) [1,7]
Many other giant viruses infecting amoeba have been isolated and described in several geographical areas worldwide, such as Pandoravirus, Pithovirus, Mollivirus, faustovirus, cedratvirus, kaumoebavirus and pacmanvirus which have recently been included into the proposed order of Megavirales [2,9,10,11,12,13,14,15]
Summary
Megavirales order has recently been proposed to replace the current nucleocytoplasmic large DNA viruses (NCLDV) based on capsid and genome sizes, ancestral genes encoding autonomy transcription and even translation components [1,2,3,4,5,6]. This order contains viruses considered giant viruses and the story began at the beginning of the 21st century with the discovery of Acanthamoeba polyphaga mimivirus (APMV) [1,7]. Several high throughput procedures were implemented to speed up the process of isolating giant virus in amoeba co-cultures and flow cytometry sorting was used to separate mixtures of these viruses and putatively identified the viral population [9,16,17,18]
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