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Correctly, the connection anywhere between stress and you may temperatures investigation throughout the heat is actually computed

We hypothesized that there would be a difference among aw conditions in glass transition temperature (Tg) for bacterial cells. Differential scanning calorimetry (DSC) is widely used as a method for measuring Tg [29,38,39]. vgl návÅ¡tÄ›vníků However, it is difficult to measure Tg of a composite using DSC because the thermogram shows intricate thermal responses . Therefore, here, thermal rheological analysis (TRA) was used to measure Tg. TRA, which measures Tg by attaching a temperature control device to a rheometer, is based on the principle of thermal mechanical analysis [28–30]. Previous studies used by TRA investigated the effect of water content on the Tg of cookies [29, 40], hazelnuts , and deep-fried food . To conduct the measurements, a sample is compressed at a temperature below Tg, and heated above Tg with compression. Then, the Tg of the sample can be determined as a force drop induced by the glass transition. This is a useful method to apply to amorphous powders. By determining Tg values, we could confirm the glass transition of bacterial cells. In addition, we sought to elucidate the influence of aw on bacterial survival and its relationship with Tg. Finally, we aimed to resolve the relationship between the state change of several Salmonella serotypes that is known to be present in low water activity foods due to glass transition and the changes in thermal resistance in a desiccation environment. The results obtained here will help to understand bacterial survival in a dry environment, which has not been clarified.

Bacterial challenges and you may culturing

Salmonella enterica Typhimurium (RMID 1985009 from the Browse Institute to have Bacterial Illness from Osaka University; remote out-of customers for the sporadic circumstances), S. enterica Chester, S. enterica Oranienburg (about Aomori Prefectural Search Research off Public Health; remote from dehydrated squid chips regarding the a break out when you look at the 1999), S. enterica Stanley (RIMD 1981001 in the Browse Institute to have Bacterial Infection out of Osaka University; separated out of clients from inside the sporadic case), and S. enterica Enteritidis (RIMD 1933001 about Lookup Institute for Bacterial Disease of Osaka University; separated of customers into the sporadic instance) were used in this research.

These serovars was indeed managed on -80°C in tryptic soya broth (TSB, Merck, Darmstadt, Germany) with which has ten% glycerol. The newest stresses were activated just after incubating during the 37°C having twenty-four h toward tryptic soy ager (TSA, Merck) plates. An isolated nest of any micro-organisms ended up being moved to 5 mL off TSB for the a good sterile centrifuge tube, incubated in the 37°C to possess 24 h, following a hundred ?L aliquot out-of cultured bacteria try added so you can eight hundred mL TSB and you will incubated at 37°C having forty eight h. The new cultured structure was in fact built-up of the centrifugation (step three,100000 ? grams, ten minute) in addition to pellets was basically resuspended in the 5 mL from uncontaminated water. Bacterial-cell pellets had been acquired by the pipetting off the extreme drinking water and you may compiled with the a vinyl plate. This new plates was indeed frozen in the -80°C to have twenty-four h ahead of drying getting 24 h playing with a freeze more dry (FDU-2200, EYELA, Tokyo, Japan). Dried microbial structure were ground, listed in an air-strict basket within wished relative moisture (% RH), that was introduced playing with soaked salt aqueous choices (43% RH: potassium carbonate, 57% RH: salt bromide, 75% RH: salt chloride, and you can 87% RH: potassium chloride), and you may held at cuatro°C to own forty-eight h. The water interest and you will temperatures floating around-tight basket was indeed consistently searched having fun with thermo recorder (TR-72wf, T and you will D, Nagano, Japan). And also the liquid interest of the bacterium is actually confirmed by the an effective h2o hobby meter (Aqualab 4TE, Decagon Devices, Arizona, USA).

Devotion from cup transition temperatures (Tg)

Thermal rheological analysis (TRA) was used to measure Tg by attaching a temperature control device to a rheometer (EZ-SX, SHIMADZU, Kyoto, Japan) (illustrated in Fig 1); the analysis is based on the principle of thermal mechanical analysis [28–30]. A dried bacterial cell sample (ca. 100 mg) was placed in the forming die (? = 3 mm) and compacted with a rheometer at ca. 10 MPa. Subsequently, the sample was compressed at ca. 5 MPa ca. for 1 to 3 min and then heated at a rate of approximately 3°C/min until the temperature reached 120°C. Pressure-time data were collected with software attached to the rheometer. In parallel, a thermocouple was attached to the bottom of the forming die and time-temperature data were collected every second using a data logger. Since pressure reduction begins at the point at which the bottom temperature of the sample reaches the mechanical Tg, the onset temperature of pressure reduction could be regarded as the Tg of the sample .

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