Strains of Giardia duodenalis from diverse parts of the world have three or four chromosomal bands in the range of 650-800 kb as defined by field inversion gel electrophoresis. The extra chromosome band in this range defined a group of strains which are geographically distinct from other strains missing this band. The cloned line WB-1B has three chromosome bands in this size range and chromosome band 3 was used to construct a library of chromosome-specific probes. In some strains examined, including BRIS/83/HEPU/106, a subset of these WB-1B probes hybridized to chromosome band 3 and to the extra chromosome band 4, indicating a partial duplication of chromosome 3 in BRIS/83/HEPU/106. This duplication was estimated to be at least 500 kb when the sizes of NotI chromosome segments which hybridized with chromosome band 3-specific probes were added. A second subset of WB-1B chromosome 3-specific probes hybridized to a fifth chromosome of strain BRIS/83/HEPU/106, in the size range 650-800 kb, which was not visible by ethidium bromide staining. The two subsets of WB-1B probes hybridized to a variety of chromosome combinations in this size range in other Giardia strains and included apparent reversal of chromosome 3 and 4 mobility as well as identification of other minor chromosomes. These data indicate that chromosome band 3 of the line WB-1B is a cluster of at least two different chromosomes that cannot be electrophoretically separated but genetic rearrangements in other strains allow separation of linkage groups carried by chromosome band 3 of WB-1B.(ABSTRACT TRUNCATED AT 250 WORDS)