New variants of Vibrio cholerae O1 biotype El Tor with attributes of the classical biotype from hospitalized patients with acute diarrhea in Bangladesh

Abstract

The sixth pandemic of cholera and, presumably, the earlier pandemics were caused by the classical biotype of Vibrio cholerae O1, which was progressively replaced by the El Tor biotype representing the seventh cholera pandemic. Although the classical biotype of V. cholerae O1 is extinct, even in southern Bangladesh, the last of the niches where this biotype prevailed, we have identified new varieties of V. cholerae O1, of the El Tor biotype with attributes of the classical biotype, from hospitalized patients with acute diarrhea in Bangladesh. Twenty-four strains of V. cholerae O1 isolated between 1991 and 1994 from hospitalized patients with acute diarrhea in Matlab, a rural area of Bangladesh, were examined for the phenotypic and genotypic traits that distinguish the two biotypes of V. cholerae O1. Standard reference strains of V. cholerae O1 belonging to the classical and El Tor biotypes were used as controls in all of the tests. The phenotypic traits commonly used to distinguish between the El Tor and classical biotypes, including polymyxin B sensitivity, chicken cell agglutination, type of tcpA and rstR genes, and restriction patterns of conserved rRNA genes (ribotypes), differentiated the 24 strains of toxigenic V. cholerae O1 into three types designated the Matlab types. Although all of the strains belonged to ribotypes that have been previously found among El Tor vibrios, type I strains had more traits of the classical biotype while type II and III strains appeared to be more like the El Tor biotype but had some classical biotype properties. These results suggest that, although the classical and El Tor biotypes have different lineages, there are possible naturally occurring genetic hybrids between the classical and El Tor biotypes that can cause cholera and thus provide new insight into the epidemiology of cholera in Bangladesh. Furthermore, the existence of such novel strains may have implications for the development of a cholera vaccine.

FIG. 1.

BglI restriction patterns of rRNA genes of V. cholerae strains compared to those of selected typical strains of the El Tor and classical biotypes of V. cholerae O1. A Southern blot of BglI-digested genomic DNA was hybridized with the 7.5-kb BamHI fragment of E. coli rRNA clone pKK3535. Lanes (including strain designations and relevant characteristics): 1, toxigenic El Tor strain G-3669 (isolated in 1969 in Bangladesh); 2 through 10, strains MH-08 (Matlab type III), MG-117159 (Matlab type III), MG-117086 (Matlab type III), MG-116926 (Matlab type III), MG-116955 (Matlab type III), MG-116025 (Matlab type III), MG-116226 (Matlab type II), MJ-1485 (Matlab type I), and MJ-1236 (Matlab type I); 11, toxigenic El Tor strain 1849 (isolated in 2001); 12, toxigenic classical biotype strain (isolated in 1963 in Bangladesh).