Nitrospira distribution, diversity, and ecophysiology were studied in hypo- and subsaline, highly alkaline lakes by amplicon sequencing, metagenomics, and cultivation-based approaches. Surprisingly, not only were Nitrospira populations detected, but they were also considerably diverse. The ability of Nitrospira enrichment cultures to oxidize nitrite at neutral to highly alkaline pH of 10.5 was demonstrated. Metagenomic analysis of a newly enriched Nitrospira species, "Candidatus Nitrospira alkalitolerans", revealed numerous adaptive features of this organism to its extreme environment.