JOUJRNAL OF BACTERIOLOGY, June 1978, p. 699-705 Vol. 134, No.

3
0021-9193/78/0134-0699$02.00/0
Copyright © 1978 American Society for Microbiology Printed in U.S.A.

Relationship Between the Heat Resistance of Spores and the

Optimum and Maximum Growth Temperatures of
Bacillus Species
A. D. WARTH
Commonwealth Scientific and Industrial Research Organization, Division of Food Research, North Ryde,
N.S.W. 2113, Australia
Received for publication 15 December 1977

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Heat resistance of spores of Bacillus strains was compared with the tempera-
ture adaptation of each strain as measured by the optimum and maximum growth
temperatures and the heat resistance of vegetative cells. Maximum growth
temperatures ranged from 31 to 760C and were little affected by the nature of the
growth medium. The temperature giving maximum growth rate was closely
correlated to the maximum temperature for growth, and about 60C lower.
Vegetative-cell heat resistance, determined on exponential-phase cells, was also
correlated with maximum growth temperature. The temperature at which spores
were inactivated with a decimal reduction time of 10 min was in the range of 75
to 1210C. This temperature was 46 ± 70C higher than the maximum growth
temperature and correlated with it and the other cell parameters. Spore heat
resistance can be considered to have two components, the temperature adaptation
characteristic of the species and the stabilization conferred by the spore state.
The very large difference in heat resistance bled data to demonstrate the generally higher
between spores of different species has often heat resistance of spores of thermophilic species
been remarked upon (17, 19). Some insights into and the lower resistance typical of psychro-
the mechanism of heat resistance in spores may trophic species.
be gained by comparison of spores having differ- The aim of the present work was to obtain
ent degrees of heat resistance. For example, quantitative data on the relationship between
analyses of spores of Bacillus strains showed a the temperature adaptation of Bacillus species
correlation between diaminopimelic acid con- and the heat resistance of their spores. Temper-
tent and heat resistance (17). It seems more than ature adaptation has been measured in three
likely, however, that differences in heat resist- ways: optimum growth temperature (OGT),
ance between species are dependent not only on maximum growth temperature (MGT), and cell
the effectiveness of the heat resistance mecha- heat resistance. Species that show higher than
nism of the spore, but also to a large degree upon average spore heat resistance with respect to the
the temperatures to which the species is temperature adaptation may have a relatively
adapted. Bacillus and Clostridium are unusual more effective protective mechanism in their
genera in the very wide range of temperatures spores and will be candidates for a study of spore
at which different species can grow. Some ther- heat resistance mechanisms.
mophilic members grow above 700C, whereas
psychrotrophs grow at 10C. In general, thermo- MATERIALS AND METHODS
philic species have much more heat-stable en- Organisms. The origins of strains used and condi-
zymes than mesophiles or psychrotrophs (4, 12, tions for growth of spores are shown in Table 1.
21). When placed in the protective environment Sporulation medium consisted of, per liter: 1.5 g of
within the spore, enzymes from thermophiles yeast extract; 1.5 g of beef extract; 5 g of peptone; and
would be expected to be relatively more heat 1.74 g of K2HPO4 (pH 7.2), to which was added 1% of
stable and thermophilic spores relatively more sporulation salts: 14.7 g of CaC2- 2H20, 10 g of
heat resistant. The tendency for thermophilic MgCl2 6H20, 2.8 g of MnSO4 .7H20, 0.5 g of
species to produce spores more resistant than ZnSO4 7H20, 0.1 g of FeSO4 7H20, and 0.1 g of
those of mesophiles is commonly observed. Ber- CuS04 * 5H20 in 1 liter of 2 mM HCI. The identity of
named species was checked by the methods of Gordon
gey (5) suggested that the great heat resistance et al. (9). Thermophilic sporeformers were isolated
of spores from thennophiles might be related to from local soil as follows. Soil (1 g) in 50 ml of sporu-
their high optimum growth temperature. More lation medium was heated for 10 min at 1000C to kill
recently, Michels and Visser (15) have assem- vegetative cells and then shaken at 60°C for 24 h. The
699
7 00 WARTH J. BAcTERioL
TABLE 1. Origin and sporulation conditions of Bacllus strains
Sporulation
Species FRR no. Origin Temp
Mediumb (0C)
B. macquariensis ATCC 23466 B683 Antarctic soil SM 15
Soil isolate 14 Local soil SM 25
Soil isolate 11 Local soil SM 27
Soil isolate 10 Local soil SM 27
Soilsolate 15 Local soil SM 25
Soil isolate 13 Local soil SM 27
Soil isolate 12 Local soil SM 27

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B. sphaericus DSM 28 J. R. Norrise G 30
B. brevis DSM 30 J. R. Norris" G 30
B. cereus subsp. mycoides B689 Laboratory SM 37
B. cereusT NRS 1417 B694 G 37
B. cerews B687 Laboratory SM 37
B. megaterium QM B1551 B690 SM 37
B. subtilis 168 B696 R G. Wake, University of Sydney SM 37
B. subtilis subsp. niger B654 SM 41
B. subtitis Porton B693 SM 45
B. subtiii B692 Heated milk SM 50
B. licheniformis B691 Unheated mik G 45
B. lichenifor"nis DSM 13 J. R. Norris" G 50
B. brevis B636 Canned cabbage G 50
B. coaguku 320 B666 Canned cabbage SP 50
B. icheniformis 2o2 SMag 50
B. licheniformis B65 University of New South Wales G 55
Soil isolate 3 LLocalsoil SM 60
Soi isolate 5 Localsoil SM 60
Soilisolate I Localsoil SM 60
B. stearothermophilu NCA 1518 B651 SM 60
B. sp BSTS University of New South Wales SM 60
B. stearothermophitus B631 Canned cabbage SM 60
B. caldolyticus B697 J. H. Lindsay SM 68
Austlian National University
a FRR denotes the culture collection at Commonwealth Scientific and Industrial Research Organization Food
Research Laboratory, North Ryde, New South Wales, Australia.
b SM, Sporulation medium (see the text); G, half-strength G medium (22); SP, soya peptone medium (14).
' Obtained from J. R. Norris, Meat Research Institute, Langford, U.K These were test organisms suppLed to
the Subcommittee on Classification of the Genus Bacilu.
reltant spore suspenson was heated for 10 min at absorbaces measured at 605 nm in a modified Perkin-
1000C and plated on sponulation agar. Three isolates Elmer model 55 spectrophotometer. At tempertures
fern dslightly in colonial morphology were used. below 500C, growth rates were measured with 20 ml
Orgnims having a lower growth temperature were of media in 100-mn side-arm flasks shaken at 200 rpm
isolated from the same soil sample. Soil in sporulatio in a covered shaking water bath. Above 50°C, 50 ml of
medium was heate for 10 min at 600C, shaken at culture was shaken in 250-ml indented flasks, and 1-
340C for 2 h to germinate mesphilic spores, heated ml samples were removed. One growing culture was
for 10 min at 60°C, and shaken at 180C for 2 days. used for growth rate determinations at three temper-
The resultant spore supe was heated for 10 min atures by successve dilution and elevation of the
at 600C and plated. Many different types of colonial temperature in seps of 2 to 3°C. Thermophle were
and cell morphology were seen among the isolates, sensitive to dilution and to temperature shock near
and six different types that grew weil at 10°C but not their MGT. Therefore dilutions were made isother-
at 400C were sltdForad strains, a spore crop was mally, and the temperature was raised over a period
grown and ued as the inoclum for all gowth and of 10 min. Rates were determined after at least five
resistance experinents. Before use, the spores were generations at the n ted temperature. MGT was
activated for 10 min at a temperature approximately also determined on solid media. Slants in screw-
200C below the temperature at which the decimal capped tubes were inoculated with heat-activated
reduction time (D) was 10 min. Vegetative growth was spores at less than the OGT, and the temperature was
on (per liter): 2.5 g of yeast extract; 5 g of tryptone; raied during the next 2 to 3 h. Tubes were incubated
and I g of glucose (pH 7.0), with apr up to 30 g/liter in a temperature gradient bar at 1C intervals or in
where nece4sry. sealed cans immersed in water baths at 2°C intervals.
OGT and MGT. Growth rates were calclted from Temperatures of all liquid and solid media were mon-
VOL. 134, 1978
itored with thermocouples during incubation. Maxi-
mum variation was ±0.20C in the gradient bar and
±0.70C in flasks and sealed cans.
Heat resistance. Spore suspensions containing 107
viable spores in 0.2 ml of 0.05 M phosphate (pH 7.0)
were heated in 1-nd thin-wailed glass ampoules. Heat
resistance of vegetative cells was determined with
growing cultures. Cultures were grown at 5 to 10°C
below the heating temperature and diluted 0.1 ml into
9.9 ml of preheated growth medium. At the required
time, samples were withdrawn; dilutions were made in
0.1% peptone, plated on nutrient agar plus 0.1% glu-
cose, and incubated at approximately 10°C below the
HEAT RESISTANCE OF SPORES 701
other hand, most mesophiles were commonly
used laboratory strains, which probably were
selected for unrelated criteria such as complete
and synchronous spore formation.
Another apparent anomaly in the selection of
cultures from laboratory collections was the de-
ficiency of strains with MGTs in the range 60 to
70°C and below 40°C (Tables 1 and 2), even
though a number of additional cultures had been
screened in an effort to fill these gaps. Additional
thermophilic and psychrotrophic strains were
therefore isolated from soil. Species with MGTs

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OGT. below 40°C were very common in soil, but none
of the thermophilic isolates had MGTs between
RESULTS 60°C and 70°C.
Choice of species. Cultures of Bacillm spe- MGT. A standard growth medium was chosen
cies were chosen from laboratory collections to after trials using nine of the bacterial strains and
be representative of a wide range of MGT. Con- 24 varieties of solid media. Addition of ferrous
sideration of the origin of these cultures, how- ethylenediaminetetraacetate (5 ,ug/ml) en-
ever, suggested a possible bias in their proper- hanced the growth of four strains near their
ties. Thermophiles were mostly isolates from MGT, and 20% sucrose stimulated one strain,
spoiled canned food and therefore had been but in only one case (B. caldolyticus) was
selected for high spore heat resistance. On the growth obtained at the next higher temperature
TABLE 2. Temperature parameters for growth and survival of vegetative cells and spores of Bacillus species
Max growth ~~MGT (OC)
Species ratea (h1 OGTh (OC) CDT' (OC) SDT (OC)
rates (h') Liquid media Solid media
B. macquariensis 0.36 25 32 33 40 88
Soil isolate 14 37 77
Soil isolate 11 1.0 28 31 37 82
Soil isolate 10 1.3 32 35 38 82
Soil isolate 15 1.3 35 38 39 89
Soil isolate 13 1.5 36 39 39
Soil isolate 12 1.5 37 41 44 92
B. cereus subsp. mycoides 1.2 33 38 41 47 89
B. cereusT 2.5 39 44 44 48 92
B. sphaericus 1.6 38 45 45 47 88
B. cereus B687 2.8 42 46 46 88
B. brevis DSM30 1.4 42 46 49 49 93
B. megaterium 2.6 42 48 50 47 89
B. subtilis subsp. niger 2.5 43 50 51 99
B. subtilis P 3.2 46 53 55 55 99
B. subtilis 168 2.9 46 54 58 97
B. subtilis B692 2.8 46 53 55 57 111
B. licheniformis B691 3.4 48 56 55 54 99
B. licheniformis DSM13 2.6 51 58 57 104
B. brevis B636 2.5 48 56 57 55 106
B. licheniformis 202 2.5 51 58 57 104
B. licheniforinis B65 3.0 49 58 60 58 103
B. coagulans B666 2.3 55 59 66 118
Soil isolate 3 4.6 66 74 74 116
Soil isolate 5 4.2 66 71 74 113
Soil isolate 1 3.9 66 71 75 113
B. stearothernophilus 1518 3.5 65 71 74 72 120
B. stearother7nophilus 8 3.5 66 74 75 71 118
B. stearothermophilus B631 4.3 66 74 76 71 118
B. caldolyticus 3.7 67 72 75 72 115
a Rate constant.
b OGT is the temperature for maximum growth rate.
CDT is the temperature at which the decimal reduction time D = 10 min.
 702 WARTH
tested (<30C) above that allowing growth on
standard medium Brain heart and other com-
plex media, variation of pH, and addition of
starch, carbon, Mg +, and Ca2+ also affected the
amount of growth but did not increase the mea-
sured MGT.
Observations of MGT in both liquid and solid
media were made on cultures which had been
adapted to high temperatures by raising the
temperature in small steps over a few hours. In
liquid media, growth rates near MGT were less
reproducible than those at lower temperatures,
and on occasions did not remain constant in one
experiment. At temperatures above MGT,
growth sometimes continued temporarily for up
to three generations after the temperature was
raised. Despite these instabilities, plots of
growth rate against temperature yielded well-
defined MGTs.
On solid media, observations of MGT were
also well defined and reproducible (±1.50C).
Consistent results were obtained for slants in-
cubated in sealed cans and in screw-capped
tubes on the temperature gradient bar. Results
for the different strains ranged from 33 to 76°C
(Table 2). With two exceptions, very similar to
slightly lower MGTs were measured using liquid
media. B. coagulans grew at 660C on solid media
but could not be grown in shake flasks above
590C. B. caklolyticus, when supplemented with
Fe , grew at 790C on solid media and 740C in
liquid media.
OGT. The temperature at which cultures
grew most rapidly was determined using the
standard growth medium. A typical result is
shown in Fig. 1. Estimates of OGT could usually
- 3
_x
7 by the observation, during determination of
z viability just above the MGT.
4 Heat resistance of spores. Spore death
0 2 < /~
0)
S. megaterium OM811551
02
30 40 50
TEMPERATURE C with an SDT above 1050C. Three of the ther-
FIG. 1. Growth rate of B. megaterium on glucose-
yeast extract-tryptone meduim. Each symbol repre-
sents a different experiment.
J. BACTERIOL.
be made to ±1.50C and are shown together with
the maximum growth rates in Table 2. OGT
clearly closely correlated with MGT and was an
average of 60C lower. No marked anomalies
between OGT and MGT were evident, and
either would be a useful indicator of the temper-
ature adaptation of species.
Heat resistance of cells. Heat inactivation
kinetics of growing cells were determined in
growth media for 16 strains. A number of the
other species were unsuitable because of exten-
sive chain formation or clumping of cells. Deci-
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mal reduction times (D) were determined at
several temperatures for each strain and where
the kinetics were not exponential, D was taken
as half the time for 99% reduction in survivors.
To facilitate comparison of growth temperatures
with heat resistance, the heat resistance data are
presented (Table 2) as a cell death temperature
(CDT), which is the temperature at which D =
10 min. As inactivation rates were very temper-
ature dependent (z = 4 to 7°C), irregularities in
inactivation rates had only small effects on the
CDT.
CDTs were mostly similar to the respective
MGTs, exceptions being the psychrotroph B.
macquariensis and B. cereus subsp. mycoides,
whose CDTs were 6 to 70C above their respec-
tive MGTs. On the other hand, in the thermo-
philic species, CDTs were a little less than the
corresponding MGTs. The lower values found in
the heat resistance assay, as compared with
growth measurement data, probably resulted
from the sensitivity of the cells to sudden dilu-
tion and temperature changes that were inher-
ent in the former technique but carefully
avoided in the latter. These values show in ther-
mophiles that, near the MGT, cell growth and
death are nearly balanced. This was confirmed
MGT of several thermophiles, of a total loss in
temperatures (SDTs) were between 82 and
1200C (Table 2) and were an average of 460C
above the respective CDTs. Most of the spore
preparations showed approximately first-order
inactivation kinetics, and viable spore counts
were greater than 30% of the microscopic spore
counts. Values of z were between 4.5 and 10.50C.
Where the semilogarithmic plot was nonlinear,
an average D value for the first two decades was
normally used. The strains having lower viabil-
ities and nonlinear inactivation kinetics were B.
mnacquariensis, B. brevis DSM30, and all strains
mophiles showed simple heat activation, and in
these cases slopes were extrapolated to zero
heating time.
VoL 134, 1978
Effect of Ca2+ treatment of spores on
their heat resistance. The heat resistance of
spores can-be altered greatly by manipulation of
their exchangeable cations (3). Recently Alder-
ton et al. (2) found that treatment of a prepara-
tion of Clostridium botulinum spores with Ca2"
greatly increased its heat resistance. These
spores, however, were grown in Ca-deficient me-
dia, and since all spore crops in the present study
were grown with Ca2+-supplemented media they
would be expected to show optimal heat resist-
ance. To test this, 10 spore preparations were
HEAT RESISTANCE OF SPORES 703
tions with SDT were near unity, suggesting that
mesophilic and thermophilic species on average
are stabilized to a similar extent in the spore
state. A plot of the distribution of SDT with
respect to MGT (Fig. 3) identifies those species
for which SDT was greater than expected from
MGT. Four species, B. macquariensis, B. sub-
tilis B692, B. coagulans, and B. stearother-
mophilus 1518, were outstanding in this respect.
DISCUSSION
The heat resistance of a spore can be consid-

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treated with Ca2" (Table 3). No changes in the
refractility of treated spores were detected, but
viable counts generally were reduced. Decimal
reduction times showed both increases and de-
creases but in no case corresponded to a change
in SDT of more than 2°C. The largest change
was with B. brevis B636. Untreated spores of
this strain showed a complex heat inactivation
curve (Fig. 2) suggestive of a mixture of sensitive
spores and more resistant, heat-activated spores.
Ca2+ treatment appears to have converted sen-
sitive spores to resistance and increased SDT by
20C.
Relationships between growth and heat
ered to be comprised of two components. One of
these is the intrinsic stability of the more labile
cellular components as expressed in vegetative
cells and in dilute aqueous media and is part of
the temperature adaptation of the species. The
other is the amount of additional stability
achieved when in the spore state. When the data
are expressed as characteristic temperatures, the
7 B. brevis
107 C

resistance parameters. The results in Table

2 show that the several temperature parameters
measured on vegetative cells give a generally 105 C

consistent indication of the temperature adap-

tation of each species. All were highly correlated 0

and each was correlated with SDT to a similar untreated

extent (Table 4). Slopes of the regression equa-
U)\ \ Ca++ treated

TABLE 3. Effect of Ca2" on spore heat resistance

0 5
Untreated Ca2"-treated' -j

Species Temp D Temp D Viability

(CC) (min) (CC) (min) loss (%)h
B. megaterium 90 6.5 89 6.5
B. cereusT 90 17 90 17
B. cereus B687 88 11 88 21 60
B. subtilis B692 110 16 113 5 30 0 20 40 60
B. licheniformis 107 5 107 5 65 MINUTES
DSM13
B. brevis B636 105 15 107 14 17 FIG. 2. Heat resistance of B. brevis B636 spores.
Soil isolate 3 116 11 116 7 70 0, Untreated spores heated at 1050C; *, Ca2+-treated
Soil isolate 5 113 11 spores heated at 107°C.
113 9 85
Soil isolate 1 113 11 113 11 7
B. stearother- 120 11 120 5 34 TABLE 4. Linear regression analysis"
mophilus 1518
B. stearother- 117 8 118 8 85 x a c r2 n
mophilus MGT, 0.88 52 0.886 28
B631 MGT, 0.854 56 0.842 28
a Spores were treated with 0.1 M calcium acetate OGT 0.89 58 0.859 28
(pH 8.7) for 2 h at 20°C above their respective MGT, CDT 1.08 41 0.918 15
washed twice, and heated in phosphate buffer (pH "SDT = ax + c, in 'C. For SDT, D = 10 min.
7.0). Abbreviations: MGT,,, MGT on solid media; MGT,,
b Reduction in unheated viable count after treat- MGT on liquid media; r , coefficient of determination;
ment with Ca2". n, number of strains.
704 WARTH
spore protective increment is expected to be
approximately additive to the intrinsic stability.
On this basis, the results show that thermophiles
on the average do not have a greater spore
protective increment than mesophiles or psy-
chrotrophs and that the wide range in heat
resistance of spores is accounted for not so much
by differences between species in the spore pro-
tective increment, which varied between 42°C
and 59°C with respect to MGT, but more as a
consequence of the very wide range of temper-
ature adaptation (over 45°C) found in Bacillus
species. A consequence of the limited natural
variation of the spore protective increment is
that the most heat-resistant spores will all be
formed by thermophiles and, conversely, unless
species are found with very much more effective
heat-protective mechanisms, that the SDT of
psychrotrophic species will be limited to about
90°C. This also provides one explanation for the
preeminence of the heat resistance of bacterial
spores, that is, that other organisms that form
spores or desiccated resting forms are not ther-
mophilic.
Several species which have high spore heat
resistance with respect to their temperature ad-
aptation have been identified in this survey.
Most of these had been isolated from heat-proc-
essed food and hence were strongly selected for
high heat resistance.
120
110 I
U
I-
49
£100
I-
x x
49
I
x x
mu
0
a.
x x
so
x
40
MAXIMUM
FIG. 3. SDT (the temperature for which D = 10 min) in relation
J. BACTERIOL.
Although species which have a higher SDT
than expected from their cell growth and sur-
vival temperatures most likely have unusually
effective spore heat-protective mechanisms,
other factors may be significant in particular
cases, i.e., where cell growth or survival is limited
by a particularly heat-sensitive process that is
not relevant to spore survival. Possible examples
are specific nutritional (8) or ion (16) require-
ments for growth or survival at high tempera-
ture. For this reason several parameters were
determined as indicators of temperature adap-
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tation. Use of each parameter nearly always
gave very similar results, indicating that special
effects were not often important. The high heat
resistance of B. macquariensis and B. cereus
subsp. mycoides cells compared with their MGT
suggests that specific factors were limiting
growth at higher temperatures. In fact, the MGT
of B. macquariensis has increased significantly
since its original isolation by Marshall and Ohye
(14). Kim and Larkin (11) reported that B. psy-
chrophilus mutated to increase OGT and MGT
by 15°C. Two other organisms, B. coagulans
and Sll, were limited in their growth in liquid
media compared with MGT on solid media. In
the other direction, species that have an effective
heat-protective mechanism may be missed if, as
reported for C. botulinum type E (1), a specific
heat sensitivity of a germination enzyme or any
x x
x
50 60 70
GROWTH TEMPERATURE C
to MGT for 28 strains of Bacillus.
VOL. 134, 1978 HEAT RESISTANCE OF SPORES 705
other specific defect lowers the SDT signifi- mostability of enzymes from BaciUus stearothermoph-
cantly. iLus and Bacillus cereus. Arch BiochenL Biophys.
125:765-769.
Murrell and Warth (17) found a correlation 5. Bergey, D. H. 1919. Thermophilic bacteria. J. Bacteriol.
between the heat resistance and diaminopimelic 4:301-306.
acid content of spores of 18 Bacilus strains. 6. Bradshaw, J. G., J. T. Peeler, and R. M. Twedt 1975.
Reexamination of their data for those strains in Heat resistance of ileal loop reactive Bacillus cereu.s
strains isolated from commercially canned food. Appl.
common with the present study shows that dia- MicrobioL 30:943-945.
minopimelate content correlates with the spore 7. Cross, T. 1968. Thermophilic actinomycetes. J. Appi.
protective component of heat resistance and to Bacteriol. 31:36-53.
a lesser extent to the temperature adaptation of 8. Garibaldi, J. A. 1971. Influence of temperature on the
iron metabolism of a fluorescent pseudomonad. J. Bac-
the species component. teriol. 105:1036-1038.
The extensive literature on heat resistance of 9. Gordon, R. E., W. C. Haynes, and C. H. Pang. 1973.

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bacterial spores generally shows the same rela- The genus Bacillus. Agriculture Handbook no. 427.
US. Agricultural Research Service, Washington, D.C.
tionships between growth temperature require- 10. Ingram, IL 1969. Sporeformers as food spoilage orga-
ments and SDT as found in this study, but the nisms, p. 549-610. In G. W. Gould and A. Hurst (ed.),
lack of precise data on temperature require- The bacterial spore. Academic Press, London.
ments for growth precludes a close comparison 11. Kim, K. J., and J. AL Larkin. 1973. Production of
in most cases. Ingram (10) and Michels and mesophilic mutants from a psychrophilic Bacillus. Can.
J. MicrobioL 19:1452-1454.
Visser (15) have compiled spore heat resistance 12. Ljungdahl, L G., and D. Sherod. 1976. Proteins from
data for psychrotrophic, mesophilic, and ther- thermophilic microorganisms, p. 147-187. In M. R.
mophilic species of Clostridum and BaciUus. Heinrich (ed.), Extreme environments. Mechanisms of
Psychrotrophic species were not well repre- 13. Mac microbial adaptation. Academic Press, London.
aiond, R. E., and S. W. MacDonaid 1962. The
sented in the present study, but data of Shehata physiology and natural relationships of the motile,
and Collins (20), Michels and Visser (15), and spore-forming Sarcinae. Can. J. Microbiol. 8:795-808.
Roberts ana Derrick (18) show that the differ- 14. Marshall, B. J., and D. F. Ohye. 1966. Bacius mac-
ences between MGT and SDT are also in the quariensis n.sp. A psychrotrophic bacterium from sub-
antarctic soil. J. Gen. Microbiol. 44:41-46.
range of 42 to 60°C. Among other spore forming 15. Michels, IL J. IL, and F. IL W. Visser. 1976. Occur-
genera, Thermoactinomyces vulgaris spores rence and thermoresistance of spores of psychrophilic
were in the normal range of heat resistance with and psychrotrophic aerobic spore-formers in soil and
SDT - MGT = 43°C (7). Sporosarcina urea 16. Mosley, foods. J. AppL Bacteriol. 41:1-11.
G. A., G. L Card, and W. L Koostra. 1976.
spores were relatively less resistant, with CDT Effect of calcium and anaerobiosis on the thermostabil-
- SDT = 34°C (13). Candidates for exceptional ity of Bacillus stearotJermophilus. Can. J. MicrobioL
spore resistance mechanism from reported data 22:468-474.
are: C. thermosaccharolyticum, which had an 17. Murreil, W. G., and A. D. Warth. 1965. Composition
and heat resistance of bacterial spores, p. 1-24. In L. L.
OGT of 54°C and an SDT of 129°C (23), and a Campbell and H. 0. Halvorson (ed.), Spores III. Amer-
mesophilic strain of B. cereus, which had a D ican Society for Microbiology, Ann Arbor, Mich.
121°C of 2.3 min and an OGT of 35°C (6). 1& Roberts, T. A., and C. IL Derrick. 1975. Sporulation of
Clostidium putrefaciens and the resistance of the
ACKNOWLEDGEMEN9T spores to heat and radiation and curing salts. J. AppL
I gratefully acknowledge the skillful technical assistance of
Bacteriol. 38:33-37.
19. Roberts, T. A., and A. D. Hitchins. 1969. Resistance of
S. Meidrum and K. Easton. spores, p. 611-670. In G. W. Gould and A. Hurst (ed.),
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