Influence of the sporulation temperature upon the heat resistance of Bacillus subtilis.

The influence of different sporulation temperatures (30, 37, 44 and 52 degrees C) upon heat resistance of Bacillus subtilis was investigated. Heat resistance was greater after higher sporulation temperatures. Relation of heat resistance and temperature of sporulation was not linear over all the range of temperatures tested. Heat resistance increased about tenfold in the range of 30-44 degrees C. Sporulation at 52 degrees C did not show any further increase in heat resistance. This effect was constant over all the range of heating temperatures tested (100-120 degrees C). z value remained constant (z = 9 degrees C). Greater heat resistances at higher temperatures of sporulation were not due to selection of more heat resistant cells by a higher sporulation temperature. Spores obtained from cells incubated at 32 or 52 degrees C always possessed heat resistances that corresponded to the sporulation temperature regardless of the incubation temperature of their vegetative cells.     

The heat resistance of ascospores of four Saccharomyces spp. isolated from spoiled heat-processed soft drinks and fruit products

The heat resistance ( D and z values) of four Saccharomyces spp., viz. S. bailii, S. cerevisiae, S. chevalieri, S. uvarum , was investigated by a previously described method (Put et al. 1977). The highest heat resistance was observed in ascospores of S. cerevisiae 175 showing a decimal reduction time or death rate constant at 60°C of 22.5 minutes. The average D 60°C values of the remaining species were: S. bailii 10 min; S. chevalieri 13 min; and S. uvarum 1.5 min; while z values varied between 4.0 and 6.5°C. It can be postulated that there may exist some comparison between the mechanism of yeast ascospore and bacterial endospore heat resistance. However, (i) washed Saccharomyces ascospores cannot be stored at 5°C for a longer period than 2–3 weeks without loss of heat resistance; and (ii) ascospore heat survival curves are essentially not linear over the whole range, which may be due to some heterogeneity of the ascospore population tested. Besides, it was observed that the heat resistance ( D 60°C values) of the Saccharomyces ascospores proved to be 50–150-fold higher than the D _60°C values of the corresponding vegetative cells.     

Influence of the incubation temperature after heat treatment upon the estimated heat resistance values of spores of Bacillus subtilis

S. CONDÓN, A. PALOP, J. RASO AND F.J. SALA. 1996. The influence of the incubation temperature on the estimated heat resistance for survivors after heat treatment was investigated. The survival curves and the D _t values of spores of Bacillus subtilis heated at different temperatures in pH 7 buffer, obtained after incubating survivors at different temperatures (30, 37, 44 or 51°C), were compared. The incubation temperature influenced the profile of survival curves. Lower incubation temperatures led to bigger D _t values and longer shoulders. D _t values obtained after incubating at 30°C were higher (x3 approx.) than those obtained by incubating at 51°C. The incubation temperature did not modify z values ( z = 9.1). These results show that shoulders are not only due to the activation of dormant spores but also to heat damage repair mechanisms. From the profile of survival curves at different incubation temperatures it would seem that heat damage is accumulative. Cells can repair the initial heat injury, but the accumulation of injuries would eventually make the damage irreversible.     

Relationship between Bacillus sphaericus spore heat resistance and sporulation temperature

Bacillus sphaericus 9602 was grown in batch culture at various temperatures. At 10°C and 12°C the maximum sporulation yield was <10%, while at 15°C, 20°C and 30°C, a sporulation yield of >95% was achieved. However at 40°C B. sphaericus grew only vegetatively. The heat resistances (D values at 90°C) of spores grown at 15°C and 20°C were significantly higher than those grown at 30°C.     

Heat resistance of Listeria monocytogenes in dairy products as affected by the growth medium

Listeria monocytogenes strains 1151 and Scott A were grown in broth at 30 °C and transferred to half cream, double cream and butter stored at 5 °C to determine the influence of dairy product composition on heat resistance at 52, 56, 60, 64 and 68 °C. Strain 1151 showed a higher heat resistance than strain Scott A. The heat resistance of both strains was higher in the dairy products than in broth, particularly at lower temperatures. A significant difference was observed between log 10 of the D -values in the different dairy products. The D -values obtained for both strains resuspended in all the dairy products would result in efficient elimination of the pathogen at 72·7 °C for 15 s. The highest D -value was 11·30 s at 68 °C and by using a z -value of 6·71 °C it can be determined that at 72·7 °C the D -value would be 1·5 s. The 15 s process would therefore achieve 10 log reductions. The effect of growth conditions on the heat resistance at 60 °C of L. monocytogenes Scott A was also investigated. When the cells were grown in the dairy products themselves, and particularly butter, the heat resistance of Scott A was enhanced; for example, the D -values were 7·15 times higher than in broth. Further studies are required to investigate if this protection against heating exists at higher temperatures, in which case the efficiency of pasteurization treatments or other heat treatments would be considerably lowered.     

Elevation of the heat resistance of Salmonella typhimurium by sublethal heat shock

The survival of Salmonella typhimurium after a standard heat challenge at 55°C for 25 min increased by several orders of magnitude when cells grown at 37°C were pre-incubated at 42°, 45° or 48°C before heating at the higher temperature. Heat resistance increased rapidly after the temperature shift, reaching near maximum levels within 30 min. Elevated heat resistance persisted for at least 10 h. Preincubation of cells at 48°C for 30 min increased their resistance to subsequent heating at 50°, 52°, 55°, 57° or 59°C. Survival curves of resistant cells were curvilinear. Estimated times for a '7D' inactivation increased by 2.6- to 20-fold compared with cells not pre-incubated before heat challenge.     

The Effects of Temperature on the Incubation and Latent Periods of Powdery Mildew (Erysiphe polygoni) on Clematis

The effects of temperature on the length of the incubation and latent periods of clematis powdery mildew, caused by Erysiphe polygoni , were studied. At constant temperatures over the range of 10–29°C, the length of the incubation period ranged from 4 to 11 days, and the length of the latent period ranged from 4 to 13 days; no visible colonies developed at 30°C after 19 days. The relationship between temperature and the rates of fungal development within the incubation and latent periods (expressed as the reciprocal of the lengths of the incubation and latent periods) under constant temperature were described well by a non-linear model. The resulting curves were not the usual form of an asymmetrically bell-shaped type; instead, they were of exponential type with the development rate increasing with increasing temperatures. The relationship between temperature and the development rate during the post-incubation (before sporulation) period was non-linear with an optimum temperature of approximately 20°C.     

Effect of culture age, pre-incubation at low temperature and pH on the thermal resistance of Aeromonas hydrophila

S. CONDON, M.L. GARCIA, A. OTERO AND F.J. SALA. 1992. The thermal resistance of Aeromonas hydrophila strain NCTC 8049 was determined within the range 48°-65°C with a thermoresistometer TR-SC and McIlvaine buffer. The effects of culture age, pre-incubation at 7°C and the pH of the heating menstruum were evaluated. The pattern of thermal death was dependent on culture age. Cells heated in the late logarithmic growth phase (15 h at 30°C) were twice as resistant as those in the early stage (5 h at 30°C), and the maximum D -value was obtained after 72 h incubation (5.5 total increase). The age of the cells did not affect z -values significantly. The heat resistance of cells incubated for 48 h at 30°C increased (twice) after holding at 7°C for 72 h. Pre-incubation at low temperature of older cultures (72 h, 30°C) did not influence their D -values. Maximum heat resistance was found at pH 6.0 and minimal at pH 4.0. Decreasing the pH from 6.0 4.0 reduced D -values by a factor of 5. Although the strain studied was heat-sensitive ( D _55°C= 0.17 min; z = 5.11°C), survivor curves of cultures older than 50 h showed a significant tailing. Organisms surviving in the tails were only slightly more resistant than were the original population.     

Heat resistance of Cronobacter species (Enterobacter sakazakii) in milk and special feeding formula

Aim:  To determine D - and z -values of Cronobacter species ( Enterobacter sakazakii ) in different reconstituted milk and special feeding formula and the effect of reconstitution of powdered milk and special feeding formula with hot water on the survival of the micro-organism.
Methods and Results:  Five Cronobacter species (four C. sakazakii isolates and C. muytjensii ) were heated in reconstituted milk or feeding formula pre-equilibrated at 52–58°C for various times or mixed with powdered milk or feeding formula prior to reconstitution with water at 60–100°C. The D -values of Cronobacter at 52–58°C were significantly higher in whole milk (22·10–0·68 min) than in low fat (15·87–0·62 min) or skim milk (15·30–0·51 min) and significantly higher in lactose-free formula (19·57–0·66 min) than in soy protein formula (17·22–0·63 min). The z -values of Cronobacter in reconstituted milk or feeding formula ranged from 4·01°C to 4·39°C. Water heated to ≥70°C and added to powdered milk and formula resulted in a > 4 log₁₀ reduction of Cronobacter .
Conclusions:  The heat resistance of Cronobacter should not allow the survival of the pathogen during normal pasteurization treatment. The use of hot water (≥70°C) during reconstitution appears to be an effective means to reduce the risk of Cronobacter in these products.
Significance and Impact of the Study:  This study supports existing data available to regulatory agencies and milk producers that recommended heat treatments are sufficient to substantially reduce risk from Cronobacter which may be present in these products.     

Thermal characterization of different isolates of Thiobacillus ferrooxidans

Abstract Ten different isolates of Thiobacillus ferrooxidans were characterized with respect to temperature in the range 2–35°C. Four of the ten strains oxidized ferrous iron exponentially over the entire range of incubation temperatures, including the lowest temperature tested (2°C), and were therefore characterized as psychrotrophic. Jarosite production was substantially reduced at temperatures less than 10°C and was not observed at 2°C. Energy of activation values were in the range 75.2–96.6 kJ/mol°C and indicated that iron oxidation at low temperatures was governed by both a chemical and a physical control.     

Effect of temperature on the hatching time of eggs of Ephemerella ignita (Poda) (Ephemeroptera:Ephemerellidae)

SUMMARY. Eggs of Ephemerella ignita (Poda) were kept at eight constant temperatures (range 5.9–19.8°C) in the laboratory. Over 85% of the eggs hatched in the temperature range 10.0–14.2°C but the percentage decreased markedly to 39% at 5.9°C and 42% at 19.8°C. Hatching time (days after oviposition) decreased with increasing water temperature over the range 5.9–14.2°C and the relationship between the two variables was well described by a hyperbola. Therefore, the time taken for development was expressed in units of degree-days above a threshold temperature. Mean values (with 95%CL) were 552 (534–573) degree-days above 4.25°C for 10% of the eggs hatched, 862 (725–1064) degree-days above 3.57°C for 50% hatched and 1383 (1294–1486) degree-days above 3.14°C for 90% hatched. These values can be used to predict hatching times at temperatures below 14.68°C for 10% hatched, 14.54°C for 50% hatched and 14.45°C for 90% hatched. At higher temperatures, the hatching time and the number of degree-days required for development both increased with increasing temperature. Equations were developed to estimate the number of degree-days required for development at these higher temperatures.
Eggs were also placed in the Wilfin Beck, a small stony stream in the English Lake District. Maximum and minimum water temperatures were recorded in each week and the summation of degree-days was used to predict the dates on which 10%, 50% and 90% of the eggs should have hatched. There was good agreement between these estimates and the actual hatching times. Only 10–15% of the eggs hatched between October and late February with most of the eggs hatching in March, April and May. Nymphs hatching in October and November probably did not survive the winter.     

Heat resistance in Salmonella enteritidis phage type 4: the influence of storage temperatures before heating

H umphrey , T.J. 1990. Heat resistance in Salmonella enteritidis phage type 4: the influence of storage temperatures before heating. Journal of Applied Bacteriology 69 493–497.
Storage of cultures of Salmonella enteritidis PT4 at either 4° or 8°C before heating significantly increased heat sensitivity. The differences between fresh and stored cultures, which became apparent after 4–7 h, were more pronounced with cultures stored at the lower temperature and in those heated at 60° rather than 55°C. Incubation of the stored cultures in either egg or Lemco broth for 30 min at 37°C prior to heating enabled the organisms to recover heat resistance.     

Heat resistance of different serovars of Listeria monocytogenes

Twelve Listeria monocytogenes strains representing seven serovars were heat-treated in physiological saline by a glass capillary tube method. Five strains were treated at 58°, 60° and 62°C, three at 60°, 62° and 64°C and four at 60°C. Heat-treated bacteria were recovered on blood agar in two ways: (1) incubation at 37°C for 7 d; and (2) preincubation at 4°C for 5 d, followed by incubation at 37°C for 7 d. D and z values were determined. Better average recovery and higher D values were obtained when the preincubation procedure was used. The final evaluations of the heat resistance properties of the strains were therefore based on values for preincubated samples. D values recorded at 58°, 60°, 62° and 64°C for preincubated samples were 1.7–3.4, 0.72–3.1, 0.30–1.3 and 0.33–0.68 min, respectively. z values determined were 5.2–6.9°C. D values were compared statistically. Significant differences in heat resistance were noted both between serovars and between strains belonging to the same serovar.     

Sporulation temperature affects initiation of germination and inactivation by high hydrostatic pressure of Bacillus cereus

The influence of sporulation temperature (20, 30 and 37 °C) on the heat resistance and initiation of germination and inactivation by high pressure on Bacillus cereus ATCC 14579 spores was investigated. Spores sporulated at 37 °C were the most heat-resistant. However, spores sporulated at 20 °C were more resistant to the initiation of germination and inactivation by high pressure. Spores were more sensitive to pressure at higher treatment temperatures. At 25 °C, there was an optimum pressure (250 MPa) for the initiation of germination for the three suspensions; at higher temperatures an increase of pressure up to 690 MPa caused progressively more germination. Resistance to the germinability and inactivation by high pressure of the spore population was distributed heterogeneously. Semilogarithmic curves of the ungerminated and survival fraction of B. cereus spores were concave. The resistant fraction of the spore population was lower at higher treatment temperatures. At 60 °C after 30 s of treatment at 690 MPa almost 5 log cycles of the population of B. cereus sporulated at 20 °C was germinated, and more than 7 log cycles of the population of B. cereus sporulated at 30 and 37 °C. The same treatment inactivated 4, 6 and 7 log cycles of the population of B. cereus sporulated at 20, 30 and 37 °C, respectively.     

Effects of sporulation pH on the heat resistance and the sporulation of Bacillus cereus

Spores of Bacillus cereus ATCC 7004, 4342 and 9818 were obtained in nutrient agar at several pH from 5·9 to 8·3. The optimum pH for sporulation was around 7, but good production of spores was obtained in the range 6·5–8·3. With all three strains, D -values clearly dropped with sporulation pH, decreasing by about 65% per pH unit. z -Values were not significantly modified ( P > 0·05) by this factor. Mean z -values of 7·13 °C ± 0·16 for strain 7004, 7·67 °C ± 0·04 for 4342 and 8·80 °C ± 0·64 for 9818 were obtained.     

Heat resistance of Campylobacter and Yersinia strains by three methods

Two methods of determining the heat resistance of bacteria, a glass cup and a test tube method, were compared with a method using capillary tubes. Three strains of Yersinia enterocolitica , one of Campylobacter jejuni and two of C. coli were tested in physiological saline. The differences between the results obtained by the glass cup method and the reference method were not statistically significant for five strains and were small also for the other, a Yersinia strain. D values obtained by the glass cup method at 58, 60 and 62°C were 1.4–1.8, 0.40–0.51 and 0.15–0.19 min ( z values 4.00–4.52°C) for the Yersinia strains, and 0.42, 0.13 and 0.07 min ( z value 5.07°C) for one C. coli strain. For the other Campylobacter strains, D values of 0.71–0.78, 0.24–0.28 and 0.12–0.14 min ( z values 4.94 and 5.60°C) were recorded at 56, 58 and 60°C. D values obtained at 60°C by the test tube method were 2.7–5.0 min and were considered to be unrealistic.     

The effects of temperature on the development and mortality of eggs of perch, Perca fluviatilis

SUMMARY. The rate of development and mortality of perch Perca fluviatilis was studied at ten different constant temperatures. The rate of development was inversely related to the incubation temperature, whereas the rate of mortality was directly related to the incubation temperature. The sum of heat (Σ H , degree-days) required for 10, 50 and 90% of the eggs to hatch was found to be constant, regardless of the incubation temperature, with mean values (with 95% confidence limits) of 91.4 (83.3–102.0) degree-days above 4.6°C for 10% hatched, 97.0 (90.9–104.2) degree-days above 4.9°C for 50% hatched and 101.0 (94.3–108.7) degree-days above 5.0°C for 90% hatched. Mortality among the different embryological stages was highest for the pre-hatching stage (i.e. when eye-pigment has been formed) at all temperatures. High mortality among the early stages occurred at temperatures below 8°C and above 12°C.     

The effect of recovery conditions on the apparent heat resistance of Bacillus cereus spores

The effect of recovery media and incubation temperature on the apparent heat resistance of three ATCC strains (4342, 7004 and 9818) of Bacillus cereus spores were studied. Nutrient Agar (NA), Tryptic Soy Agar (TSA), Plate Count Agar (PCA) and Milk Agar (MA) as the media and temperatures in the range of 15–40°C were used to recover heated spores. Higher counts of heat injured spores were obtained on PCA and NA. The optimum subculture temperature was about 5°C below the optimum temperature for unheated spores. No significant differences in heat resistance were observed with the different recovery conditions except for strains 4342 and 9818 when MA was used as plating medium.
Large differences in D -values were found among the strains ( D ₁₀₀=0·28 min for 7004; D ₁₀₀=0·99 min for 4342; D ₁₀₀= 4·57 min for 9818). The 7004 strain showed a sub-population with a greater heat resistance. The z values obtained for the three strains studied under the different recovery conditions were similar (7·64°C 0·25).     

The effect of temperature on the ability of Tilapia mossambica Peters to enter deep water

The ability of adult Tilapia mossambica Peters to enter deep water was determined at 15, 22 and 30°C. At 30°C adults compensate to about 20m depth but at 15°C to only 7 m. Compensation is more rapid at high than at low temperatures. T. mossambica haemoglobin has a marked Root effect which is the same at 22 and 30°C. The oxygen affinity of the haemoglobin is higher at 15°C than at 30°C. There was no measurable difference in the rate of passive oxygen diffusion across the swimbladder wall in the temperature range 15–30°C. It is concluded that the ability to enter deeper water at higher temperatures is related to decreased oxygen affinity of the haemoglobin and higher rates of oxygen secretion and blood circulation.     

Elevated red myotomal muscle temperatures in the most basal tuna species, Allothunnus fallai

The present study tested the hypothesis that Allothunnus fallai can elevate its slow-oxidative red myotomal muscle (RM) temperature. Measurements on 30 A. fallai (750–850 mm fork length) captured by hook and line off the coast of southern New Zealand revealed that RM temperatures are elevated by mean ± s . d . 8·1 ± 1·3° C (range 6·7–10·0° C) above the mean ± s . d . ambient sea surface temperature 15·3 ± 0·8° C (range 14·3 to 16·4° C). These data provide evidence that the vascular modifications to the central circulation of A. fallai act as a counter-current heat exchanger and that RM heat conservation is a character state present in all extant tuna species.