Open AccessResuscitation of Vibrio cholerae O1 strain TSI‐4 from a viable but nonculturable state by heat shock
Author(s) -
Wai Sun Nyunt,
Moriya Tetsuhiro,
Kondo Katsuhiko,
Misumi Hiroyasu,
Amako Kazunobu
Publication year - 1996
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1996.tb08047.x
Subject(s) - viable but nonculturable , microbiology and biotechnology , nutrient agar , bacteria , vibrio cholerae , strain (injury) , agar , shock (circulatory) , biology , agar plate , vibrionaceae , yeast extract , chemistry , anatomy , medicine , genetics
Vibrio cholerae strain TSI‐4 was incubated in an M9 salt solution at 15 °C for more than 100 days. The plate counts showed no viable cells on day 30, but a broth culture from that day showed the growth of bacteria. However, after 35 days the bacteria entered the nonculturable state, based on the assessment of both the plate counts and broth culture. A portion of the culture was heated at 45 °C for 1 min in a water bath and subsequently plated onto a nutrient agar plate. More than 1000 colonies were recovered after this heat‐shock treatment. The recovered cells showed the same chromosomal DNA pattern in the restriction map and the same outer membrane protein pattern in SDS‐PAGE. Recovery of viable cells by heat‐shock was achieved in cultures grown on M9 salt but not from cultures grown in phosphate‐buffered saline. This suggests that the presence of NH 4 Cl in the M9 salt solution may support the growth of the bacteria in a low nutrient medium, while also playing an important role in resuscitation.