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Two actinomycete strains, designated H225(T) and H137, were isolated from two soil samples collected from the arid region of Ahbas at Beni-Isguen (Mzab), located in the Algerian Sahara. Phylogenetic analyses based on 16S rRNA gene sequences indicated that the novel strains should be assigned to the genus Prauserella of the family Pseudonocardiaceae, and they were therefore subjected to a polyphasic taxonomic study. These two strains contained meso-diaminopimelic acid as the diagnostic diamino acid and arabinose and galactose as major wholecell sugars. The diagnostic phospholipid was phosphatidylethanolamine. The predominant menaquinone was MK-9(H-4), and the major fatty acid was iso-C-16:0. DNA-DNA hybridization values between strain H225T and its closest phylogenetic neighbours, namely Prauserella flava DSM 45265(T), Prauserella alba DSM 44590(T), Prauserella aidingensis DSM 45266(T), Prauserella salsuginis DSM 45264(T) and Prauserella sediminis DSM 45267(T), were clearly below the 70% threshold used for species delineation. The genomic DNA G+C content of strains H225(T) and H137 was 70.4 mol%. On the basis of phenotypic and genotypic data, strains H225(T) and H137(T) are considered to represent a novel species of the genus Prauserella, for which the name Prauserella isguenensis sp. nov. is proposed. The type strain is H225(T) (=DSM 46664(T)=CECT 8577(T)).