Cytogenetic analysis of the larval polytene chromosomes of Anopheles nuneztovari from 5 collection sites in Tachira and Zulia states northwest of the Andean Cordillera in western Venezuela and from 2 sites in the Department of Valle, western Colombia, revealed what appears to be a distinctive cytotype informally designated as An. nuneztovari C. Its chromosomes are homosequential with those of An. nuneztovari B from western Venezuela southeast of the Cordillera but differ in the presence of a well-defined chromocenter and unique inversion polymorphisms. The large complex inversion in western Venezuela, 2Lb, is present at a frequency of 0.263 and deviates significantly from Hardy-Weinberg equilibrium in 3 of the 5 sites. Two smaller inversions (2Lc and 2Ld) that are included in 2Lb are present in the Colombian samples at a frequency of 0.300.