Dataset Information

Relationship between glass transition temperature, and desiccation and heat tolerance in Salmonella enterica.

ABSTRACT: Pathogenic bacteria such as Salmonella enterica exhibit high desiccation tolerance, enabling long-term survival in low water activity (aw) environments. Although there are many reports on the effects of low aw on bacterial survival, the mechanism by which bacteria acquire desiccation tolerance and resistance to heat inactivation in low-aw foods remains unclear. We focused on the glass transition phenomenon, as bacteria may acquire environmental tolerance by state change due to glass transition. In this study, we determined the glass transition temperature (Tg) in S. enterica serovars under different aw conditions using thermal rheological analysis (TRA). The softening behaviour associated with the state change of bacterial cells was confirmed by TRA, and Tg was determined from the softening behaviour. Tg increased as the aw decreased in all S. enterica serovars. For example, while the Tg of five S. enterica serovars was determined as 35.16°C to 57.46°C at 0.87 aw, the Tg of all the five serovars increased by 77.10°C to 83.30°C at 0.43 aw. Furthermore, to verify the thermal tolerance of bacterial cells, a thermal inactivation assay was conducted at 60°C for 10 min under each aw condition. A higher survival ratio was observed as aw decreased; this represented an increase in Tg for Salmonella strains. These results suggest that the glass transition phenomenon of bacterial cells would associate with environmental tolerance.

SUBMITTER: Lee K

PROVIDER: S-EPMC7259547 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature

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Relationship between glass transition temperature, and desiccation and heat tolerance in Salmonella enterica.

Lee Kyeongmin K Shoda Masaki M Kawai Kiyoshi K Koseki Shigenobu S

PloS one 20200529 5

Pathogenic bacteria such as Salmonella enterica exhibit high desiccation tolerance, enabling long-term survival in low water activity (aw) environments. Although there are many reports on the effects of low aw on bacterial survival, the mechanism by which bacteria acquire desiccation tolerance and resistance to heat inactivation in low-aw foods remains unclear. We focused on the glass transition phenomenon, as bacteria may acquire environmental tolerance by state change due to glass transition. ...[more]

PMID: 32469949

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Project description:Recent heightened interest in inhaled bacteriophage (phage) therapy for combating antibacterial resistance in pulmonary infections has led to the development of phage powder formulations. Although phages have been successfully bioengineered into inhalable powders with preserved bioactivity, the stabilization mechanism is yet unknown. This paper reports the first study investigating the stabilization mechanism for phages in these powders. Proteins and other biologics are known to be preserved in dry state within a glassy sugar matrix at storage temperatures (T s) at least ~50°C below the glass transition temperature (T g). This is because at (T g - T s) >50°C, molecules are sufficiently immobilized with reduced reactivity. We hypothesized that this glass stabilization mechanism may also be applicable to phages comprising mostly of proteins. In this study, spray dried powders of Pseudomonas phage PEV20 containing lactose and leucine as excipients were stored at 5, 25 or 50°C and 15 or 33% relative humidity (RH), followed by assessment of bioactivity (PEV20 stability) and physical properties. PEV20 was stable with negligible titer loss after storage at 5°C/15% RH for 250?days, while storage at 33% RH caused increased titer losses of 1 log10 and 3 log10 at 5 and 25°C, respectively. The plasticizing effect of water at 33% RH lowered the T g by 30°C, thus narrowing the gap between T s and T g to 19-28°C, which was insufficient for glass stabilization. In contrast, the (T g - T s) values were higher (range, 46-65°C) under the drier condition of 15% RH, resulting in the improved stability which corroborated with the vitrification hypothesis. Furthermore, phage remained stable (?1 log10) when the (T g - T s) value lay between 26-48°C, but became inactivated as the value fell below 20°C. In conclusion, this study demonstrated that phage can be sufficiently stabilized in spray dried powders by keeping the (T g - T s) value above 46°C, thus supporting the vitrification hypothesis that phages are stabilized by immobilization inside a rigid glassy sugar matrix. These findings provide a guide to better manufacture and storage practices of inhaled phage powder products using for translational medicines.

Dataset Information

Relationship between glass transition temperature, and desiccation and heat tolerance in Salmonella enterica.

Publications

Relationship between glass transition temperature, and desiccation and heat tolerance in Salmonella enterica.

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