Abstract:Dormant spores of Bacillus megaterium were activated for germination on glucose by heating them in aqueous suspension (but not if heated dry), by treating them with aqueous ethyl alcohol at 30 C, or by exposing them to water vapor at room temperature. The degree of water vapor activation depended upon the relative humidity, the time, and the temperature of exposure. Activation increased the extent and rate of glucose-induced germination and decreased the average microlag. Extended water vapor treatment also ac… Show more
“…Thus, the alteration of the germination rate might occur prior to, or in the early stage of germination. The germination rates of the heated spores remained constant for more than one month when they were stored at -40 C, 4 C, or 20 C. A small amount of dipicolinic acid was released during heat treatment of the spores of B. subtilis as reported in B. cereus (9) and B. megaterium (7,14,15). The amount of dipicolinic acid in the exudate of the spores treated at 65 C for 30 min was about 6% of that in the exudate at 120 C for 20 min.…”
mentioning
confidence: 93%
“…Many investigators have analyzed the effects of heat treatment on the germination response of the spores of various species and strains (2)(3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17). Effects of heat (i) on the dormant state prior to germination, (ii) on the trigger reaction of germination, and (iii) on the later events during germination are discussed below.…”
mentioning
confidence: 99%
“…The following observations may be associated with the first point. It is generally postulated that water is essential for spore activation, and heating alters the molecular structure of liquid water, so that it can readily penetrate and hydrate a critical spore site for germination (7,11). Heat also changes reversibly the tertiary structure of a protein responsible for the maintenance of the dormant state of the B. cereu s spore, resulting in a change in the permeability (8).…”
Sublethal preheating of bacterial spores has been used to stimulate their germination. This treatment is known as "heat activation" or "heat shock." In our germination experiments of spores of Bacillus subtilis PCI219 (18-23), a sufficient and reproducible germination rate could be obtained with L-alanine and its analogs without preheating, and the heated spores required higher concentrations of Lalanine to germinate than the unheated spores in the presence of glucose. In the present study we examined the effect of heat treatment on the germination response, especially on the sites for the germinant and related compounds in the alanine receptor field of the spores (22). We found that heat treatment at 65 C for 30 min did not alter the binding sites for L-alanine, D-alanine, and the hydrophobic compounds, but it did affect the binding site for glucose. The results are described in some detail as follows.Actually, heat treatment of the spores of B. subtilis at various temperatures altered the germination rate (Fig. 1). At a sufficient concentration (1 mm) of L-alanine (21) the maximum germination rate was obtained by preheating at around 65 C for 30 min with or without glucose. The maximum germination rate of heated spores on L-alanine (Fig. 1A) or L-alanine plus glucose (Fig. 1B) was about 1.4 times more than that of the respective unheated ones, showing the heat activation. Heating at 70 C and higher temperatures caused a rapid decrease in germination rate. However, at lower concentrations of L-alanine only a little stimulation of the rate was observed between 50 and 60 C with L-alanine alone (Fig. 1A), and the germination rate decreased gradually as the preheating temperature in creased with L-alanine plus glucose (Fig. 1B). Germinability on glucose alone was not significant (Fig. 1B). These different responses of the germination rate to temperature indicated the complex effects of preheating.Preheating spores at 65 C for 30 min, a condition at which the maximum germination rate was obtained, was used in subsequent experiments. In the concentration-germination response curves the level of the maximum germination rate of heated spores in the absence of glucose was higher than that of unheated ones, but the apparent affinities of both spores for L-alanine, which were determined as the concentration of L-alanine showing 50 % of the maximum rate of each curve, were 101 1
“…Thus, the alteration of the germination rate might occur prior to, or in the early stage of germination. The germination rates of the heated spores remained constant for more than one month when they were stored at -40 C, 4 C, or 20 C. A small amount of dipicolinic acid was released during heat treatment of the spores of B. subtilis as reported in B. cereus (9) and B. megaterium (7,14,15). The amount of dipicolinic acid in the exudate of the spores treated at 65 C for 30 min was about 6% of that in the exudate at 120 C for 20 min.…”
mentioning
confidence: 93%
“…Many investigators have analyzed the effects of heat treatment on the germination response of the spores of various species and strains (2)(3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17). Effects of heat (i) on the dormant state prior to germination, (ii) on the trigger reaction of germination, and (iii) on the later events during germination are discussed below.…”
mentioning
confidence: 99%
“…The following observations may be associated with the first point. It is generally postulated that water is essential for spore activation, and heating alters the molecular structure of liquid water, so that it can readily penetrate and hydrate a critical spore site for germination (7,11). Heat also changes reversibly the tertiary structure of a protein responsible for the maintenance of the dormant state of the B. cereu s spore, resulting in a change in the permeability (8).…”
Sublethal preheating of bacterial spores has been used to stimulate their germination. This treatment is known as "heat activation" or "heat shock." In our germination experiments of spores of Bacillus subtilis PCI219 (18-23), a sufficient and reproducible germination rate could be obtained with L-alanine and its analogs without preheating, and the heated spores required higher concentrations of Lalanine to germinate than the unheated spores in the presence of glucose. In the present study we examined the effect of heat treatment on the germination response, especially on the sites for the germinant and related compounds in the alanine receptor field of the spores (22). We found that heat treatment at 65 C for 30 min did not alter the binding sites for L-alanine, D-alanine, and the hydrophobic compounds, but it did affect the binding site for glucose. The results are described in some detail as follows.Actually, heat treatment of the spores of B. subtilis at various temperatures altered the germination rate (Fig. 1). At a sufficient concentration (1 mm) of L-alanine (21) the maximum germination rate was obtained by preheating at around 65 C for 30 min with or without glucose. The maximum germination rate of heated spores on L-alanine (Fig. 1A) or L-alanine plus glucose (Fig. 1B) was about 1.4 times more than that of the respective unheated ones, showing the heat activation. Heating at 70 C and higher temperatures caused a rapid decrease in germination rate. However, at lower concentrations of L-alanine only a little stimulation of the rate was observed between 50 and 60 C with L-alanine alone (Fig. 1A), and the germination rate decreased gradually as the preheating temperature in creased with L-alanine plus glucose (Fig. 1B). Germinability on glucose alone was not significant (Fig. 1B). These different responses of the germination rate to temperature indicated the complex effects of preheating.Preheating spores at 65 C for 30 min, a condition at which the maximum germination rate was obtained, was used in subsequent experiments. In the concentration-germination response curves the level of the maximum germination rate of heated spores in the absence of glucose was higher than that of unheated ones, but the apparent affinities of both spores for L-alanine, which were determined as the concentration of L-alanine showing 50 % of the maximum rate of each curve, were 101 1
“…Heat activation has been credited with increasing the germination rate, activating enzymes which are dormant in resting spores, and broadening the variety of agents that will induce initiation (Keynan, Issahary-Brand & Evenchick, 1965). The activation requirements may vary depending on the sporulation medium, the chemical composition of the spore, its storage history, and the nature of the germination agent (Hyatt & Levinson, 1968). It has been postulated that the site of activation is an enzyme or enzyme system that is activated by hydration as the result of treatments which change the structure of water t o facilitate its access to the site (Hyatt & Levinson, 1968).…”
Section: Effect Of Curing Salts On Heat Activation Initiation Of Germentioning
confidence: 99%
“…The first, which is not necessary for all species (Keynan, Issahary-Brand & Evenchik, 1965), is activation. Activation is considered to be a reversible process that conditions the spore for germination (Hyatt & Levinson, 1968) The second stage is initiation of germination which terminates irreversibly the cryptobiotic state and results in a viable spore that lacks the typical resistance seen during dormancy (Sierra & Bowman, 1969). The third stage of development, occurring subsequently to initiation, has been called outgrowth This step depends on the synthesis of new macromolecules for development of the spore into a vegetative cell (Halvorson, Vary & Steinberg, 1966).…”
Von endotroph in Tris-Puffer mit Calcium-, Magnesium-und Manganionen beeinfluDten Bacillus stearothermophilus-Sporen wurden fur eine reine Dampferhitzung bei 121 "C die uberlebensdaten aufgenommen. Unter vergleichbaren Bedingungen erfolgte nach der HALVORSONund Zmormc-Gleichung beim Einsatz von 24 Cellulosescheiben rnit angetrockneten Sporen der letztmogliche Uberlebensnachweis fur Ca-Sporen nach 9 min bei I>,,, = 1,6 min, fur Mg-Sporen nach 7,2 min bei Dial : 1,3 min und fur Mnund Tris-Sporen nach 5,5 nrin bei D,,, = 1,0 min. Aus 9 verschiedenen Uberlebenskurven konnte von angetrockneten Ca-Sporen eine Absterbekurve in dem Temperaturbereich 116-135% mit einen z-Wert von 10 "C ermittelt werden.Entscheidenden EinfluB auf dic Hitzeresistenz von Bakteriensporen haben im Zusammenhang mit der Dipicolinsiiure Calcium-und evtl. weitere bivalente Kationen. Mit stcigcndem Calcium-und Dipicolinsiiuregehalt erhoht sich die Hitzeresistenz, wobei angenommen wird, daD beide Verbindungen unter Bcteiligung von Aminosliurcn bzw. Peptiden eine besondere Chelatbindung ein-
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