The present invention discloses new live attenuated strains for oral
immunization against cholera that are provided in freeze dried
formulations for long term storage and administration to humans. These
strains combine the two most important properties of live attenuated
cholera vaccine candidates. One such property is being well tolerated by
people ingesting them. This was achieved by virtue of mutations already
described in the art. The second property is having enhanced
environmental safety due to the absence of VGJ.PHI. DNA in their genomes
and also due to null mutations in the mshA gene or other spontaneous
mutations conducive to the lack of MSHA type IV fimbria on the bacterial
surface. This was done envisioning that VGJ.PHI. is a filamentous phage
able to recombine with CTX.PHI. and disseminate the cholera toxin genes.
This VGJ.PHI. phage as well as the VGJ.PHI.-CTX.PHI. recombinants uses
the MSHA fibers as receptor. Being devoid of MSHA fimbria the vaccine
candidates are protected from acquiring CTX.PHI. from the recombinant
hybrid VGJ.PHI.-CTX.PHI.. Being devoid of VGJ.PHI., the vaccine
candidates are impaired in the dissemination of CTX.PHI., via VGJ.PHI..