Dodecylamine rapidly kills of spores of multiple Firmicute species: properties of the killed spores and the mechanism of the killing.

AIMS: Previous work showed that Bacillus subtilis dormant spore killing and germination by dodecylamine take place by different mechanisms. This new work aimed to optimize killing of B. subtilis and other Firmicutes spores and to determine the mechanism of the killing. METHODS AND RESULTS: Spores of seven Firmicute species were killed rapidly by dodecylamine under optimal conditions and more slowly by decylamine or tetradecylamine. The killed spores were not recovered by additions to recovery media, and some of the killed spores subsequently germinated, all indicating that dodecylamine-killed spores truly are dead. Spores of two species treated with dodecylamine were more sensitive to killing by a subsequent heat treatment, and spore killing of at least one species was faster with chemically decoated spores. The cores of dodecylamine-killed spores were stained by the nucleic acid stain propidium iodide, and dodecylamine-killed wild-type and germination-deficient spores released their stores of phosphate-containing small molecules. CONCLUSIONS: This work indicates that dodecylamine is likely a universal sporicide for Firmicute species, and it kills spores by damaging their inner membrane, with attendant loss of this membrane as a permeability barrier. SIGNIFICANCE AND IMPACT OF THE STUDY: There is a significant need for agents that can effectively kill spores of a number of Firmicute species, especially in wide area decontamination. Dodecylamine appears to be a universal sporicide with a novel mechanism of action, and this or some comparable molecule could be useful in wide area spore decontamination.

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. SIGNIFICANCE AND IMPACT OF THE STUDY: Given potential deleterious effects of spores of Bacillales and Clostridiales, there is an ongoing interest in new ways of spore killing. A recent paper (mSphere 3: e00597-1, 2018) reported that glycerol monolaurate (GML) in a non-aqueous gel (GMLg) effectively killed spores of many species. We now find that (i) the Bacillus subtilis spores prepared as in the previous report were impure and (ii) GMLg gave no killing of purified spores of Bacillales and Clostridiales species in ≤5 h under the published conditions. Thus, GMLg is not an effective sporicide, though may prevent spore germination or kill germinated spores.

Oxygen transfer and evolution in microalgal culture in open raceways.

The mass transfer characteristics of all sections of a 100 m(2) raceway were evaluated. The efficiency of different diffusers was determined dynamically and the most effective was used for steady state system characterisation at water depth 0.2 m and velocity 0.22 m s(-1). Mass transfer coefficients at a gas flow rate of 6 m(3) h(-1) were 164.50, 63.66, 0.87 and 0.94 h(-1) for the paddlewheel, sump, straight and curved channel sections, with associated oxygen transfer rates of 106, 172, 27 and 39 g h(-1). Oxygen supersaturation during algal cultivation led to a reduction in biomass productivity, which was more severe with pure CO2 than flue gas. Simulations showed the energy required to increase mass transfer and reduce oxygen concentrations was more than compensated for by increased biomass and potential energy yields. Oxygen removal is likely to be a critical criterion, and maintenance of mass transfer by sparging may be necessary even when CO2 is not required.

Evaluation of flocculants for the recovery of freshwater microalgae.

The use flocculants on the recovery of freshwater microalgae is studied. Flocculants tested include metal salts, chitosan, and polyelectrolytes used in wastewater treatment processes. Influence of flocculant, but also the doses and biomass concentrations affecting biomass recovery as well as the concentration factor has been evaluated. Results showed that the use of metal salts or chitosan was not efficient, whereas polyelectrolytes allow the efficient recovery of biomass, at doses of 2-25 mg per gram of microalgae biomass. The required doses depend on the particular polyelectrolyte and the freshwater strain present; but cationic polyelectrolytes are generally recommended. The use of polyelectrolytes does not adversely affect water reuse in the production process. The concentration factors obtained are higher than 35 in most cases. Such high concentration factors allow a reduction in the equipment size necessary for biomass dewatering, thus improving the viability of using these microorganisms in biofuel or wastewater processes.