Lack of efficient killing of purified dormant spores of Bacillales and Clostridiales species by glycerol monolaurate in a non‐aqueous gel

Inactivation of Bacillales and Clostridiales spores is of interest, since some cause food spoilage and human diseases. A recent publication (mSphere 3: e00597‐1, 2018) reported that glycerol monolaurate (GML) in a non‐aqueous gel (GMLg) effectively killed spores of Bacillus subtilis, Bacillus cereus and Clostridioides difficile, and Bacillus anthracis spores to a lesser extent. We now show that (i) the B. subtilis spores prepared as in the prior work were impure; (ii) if spore viability was measured by diluting spores 1/10 in GMLg, serially diluting incubations 10‐fold and spotting aliquots on recovery plates, there was no colony formation from the 1/10 to 1/1000 dilutions due to GMLg carryover, although thorough ethanol washes of incubated spores eliminated this problem and (iii) GMLg did not kill highly purified spores of B. subtilis, B. cereus, Bacillus megaterium and C. difficile in 3–20 h in the conditions used in the recent publication. GMLg also gave no killing of crude B. subtilis spores prepared as in the recent publication in 5 h but gave ~1·5 log killing at 24 h. Thus, GMLg does not appear to be an effective sporicide, although the gel likely inhibits spore germination and could kill spores somewhat upon long incubations.

[1]  G. Korza,et al.  Killing of spores of Bacillus species by cetyltrimethylammonium bromide , 2019, Journal of applied microbiology.

[2]  P. Schlievert,et al.  Glycerol Monolaurate (GML) and a Nonaqueous Five-Percent GML Gel Kill Bacillus and Clostridium Spores , 2018, mSphere.

[3]  P. Setlow Observations on research with spores of Bacillales and Clostridiales species , 2018, Journal of applied microbiology.

[4]  R. Raffin,et al.  Evaluation of antimicrobial activity of glycerol monolaurate nanocapsules against American foulbrood disease agent and toxicity on bees. , 2016, Microbial pathogenesis.

[5]  A. Shen,et al.  Characterization of Clostridium difficile Spores Lacking Either SpoVAC or Dipicolinic Acid Synthetase , 2016, Journal of bacteriology.

[6]  W. H. Coleman,et al.  Mechanism of killing of spores of Bacillus cereus and Bacillus megaterium by wet heat , 2010, Letters in applied microbiology.

[7]  P. Setlow Spores of Bacillus subtilis: their resistance to and killing by radiation, heat and chemicals , 2006, Journal of applied microbiology.

[8]  P. Setlow,et al.  Treatment with oxidizing agents damages the inner membrane of spores of Bacillus subtilis and sensitizes spores to subsequent stress , 2004, Journal of applied microbiology.

[9]  P. Setlow,et al.  Lipids in the inner membrane of dormant spores of Bacillus species are largely immobile. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[10]  D. Hoover,et al.  Bacterial spore inhibition and inactivation in foods by pressure, chemical preservatives, and mild heat. , 2000, Journal of food protection.

[11]  Peter Setlow,et al.  Characterization of Spores of Bacillus subtilis Which Lack Dipicolinic Acid , 2000, Journal of bacteriology.

[12]  M. Linder,et al.  Inhibition of Bacillus licheniformis spore growth in milk by nisin, monolaurin, and pH combinations , 1999, Journal of applied microbiology.

[13]  A. Moir,et al.  Role of the gerI Operon ofBacillus cereus 569 in the Response of Spores to Germinants , 1998, Journal of bacteriology.

[14]  F. Busta,et al.  Inhibition by Monoglycerides of l-Alanine-Triggered Bacillus cereus and Clostridium botulinum Spore Germination and Outgrowth. , 1996, Journal of food protection.

[15]  F. Busta,et al.  Inhibition of Bacterial Spores and Vegetative Cells by Glycerides. , 1996, Journal of food protection.

[16]  P. Setlow,et al.  Role of DNA repair in Bacillus subtilis spore resistance , 1996, Journal of bacteriology.

[17]  J. Foster,et al.  GERMINATION OF BACTERIAL SPORES WITH ALKYL PRIMARY AMINES , 1961, Journal of bacteriology.

[18]  P. Setlow,et al.  Spores and Their Significance , 2019, Food Microbiology.