Journal of Food Engineering 80 (2007) 402–407

www.elsevier.com/locate/jfoodeng

Preparation of freeze-dried yoghurt as a space food

Elena Venir a, Manuela Del Torre a, Mara Lucia Stecchini a,*
,
Enrico Maltini a, Paolo Di Nardo b
a
Department of Food Science, University of Udine, Via Marangoni 97, 33100 Udine, Italy
b
Centro di Bio-medicina Spaziale, Università di Roma ‘‘Tor Vergata’’, Via O. Raimondo, 00173 Roma, Italy

Received 13 October 2005; accepted 20 February 2006

Available online 9 November 2006

Abstract

To develop a satisfying, calcium-rich food for consumption in space, white skim yoghurt, both plain and with added blueberries, was
subjected to freeze-drying. Optimised freeze-drying parameters were extracted from the thermal analysis of the yoghurts, taking into
account the role of the added ingredients. Rheological properties were not seriously affected by freezing, while the drying step resulted
in an overall structural weakening of the reconstituted products, possibly as a consequence of the mechanical energy required for mixing
with water. However, its viscoelastic properties were retained and the original strength could be recovered by modulating the amount of
water. The freeze-drying process affected survival of the lactic acid bacteria, resulting in a 2–3 log population reduction. Mortality levels
were reduced when sucrose and blueberries were added as ingredients.
Ó 2006 Elsevier Ltd. All rights reserved.

Keywords: Space food; Freeze-drying; Yoghurt; Functional properties; Texture; Lactic-acid bacteria

1. Introduction (Smith & Heer, 2002). The food system should provide
not only the nutrients needed for the astronauts’ survival
The Italian space agency (ASI) has been involved for but also enhance the well-being of the crew by being a
some years in an extended research program covering fun- familiar element in an unfamiliar and hostile environment.
damental aspects of human life in space travel. Human Appetizing foods can play a primary role in reducing the
nutrition in space is an important part of the program. stress of prolonged space missions and especially since it
The stress factors involved in space flight, including appears that microgravity might be responsible for affect-
microgravity, isolation, radiation, time demands and ing chemosensory perception, resulting in taste and odours
diminished resistive physical activity, can have a variety alterations in space (Olabi, Lawless, Hunter, Levitsky, &
of negative consequences including bone loss and muscle Halpern, 2002). Thus, palatable, enjoyable meals have been
wasting (Lupton & Turner, 2002). Research suggests that considered, from an astronaut’s perspective, to be an
microgravity-related bone loss is associated with both important factor in enduring the weightlessness, isolation
increases in resorption and decreases in bone formation and dangers of long space flights (Kerwin & Seddon, 2002).
(Vodovotz, Smith, & Lane, 2000). Deficient nutrient The demand for pleasant, calcium-rich food for space
intake, especially energy, calcium and vitamins D and K, may be complicated by problems arising from processing
may affect bone metabolism (Heer, 2002). Ensuring ade- conditions, shelf-life and bioavailability.
quate nutrient intake is therefore essential in order not to Within the context described above, it is likely that a
exacerbate bone loss in space caused by malnutrition yoghurt preparation would be an appropriate item to
include in a space menu.
*
Corresponding author. Tel.: +39 432 590711; fax: +39 432 590719. The nutritional and therapeutic effects of yoghurt are
E-mail address: [email protected] (M.L. Stecchini). well known and mainly attributed to fermentative changes

0260-8774/$ - see front matter Ó 2006 Elsevier Ltd. All rights reserved.
doi:10.1016/j.jfoodeng.2006.02.030
 E. Venir et al. / Journal of Food Engineering 80 (2007) 402–407 403

in the milk and/or the metabolic effects of the yoghurt of freeze-dried samples were hermetically sealed in a nitro-
microflora. In addition, it may be possible to induce greater gen atmosphere (Audionvac, Gandus, Milan, Italy) in plas-
versatility in its sensorial properties by controlling texture tic bags, and stored at 4 °C up to 45 days, a period
and by the addition of ingredients such as fruit juices or consistent with a short space mission. It was considered
fruit pieces, sweeteners, and flavours. Moreover, yoghurt that for a freeze-dried biologically active material, long-
is traditionally associated with a healthy image in both term storage would be better performed at freezing temper-
eastern and western cultures. Raspberry yoghurt has atures. Samples were reconstituted by gentle mixing with
already been included in the Shuttle menu (Perchonok & water, to reduce textural stresses, up to hydration levels
Bourland, 2002). ranging from 30% to 100% of the initial water content.
The freeze-drying process is an elective one for space
foods, as it allows nutritional and sensorial qualities to 2.3. Thermal analyses
be retained, together with extreme reduction in weight,
high solubility, long shelf life at moderate temperature Thermal analyses were used to investigate the thermal
and the possibility to perform rehydration at any desired behaviour of anhydrous and reconstituted yoghurt sam-
level. Studies on the freeze-drying of yoghurt have been ples. The anhydrous yoghurt samples were obtained
published (Sharma, Arora, & Mital, 1992; Sharma & through dehydration of freeze-dried samples over P2O5
Arora, 1995), showing the influence of process parameters up to constant weight. Glass transitions (Tg) and initial
and milk solid concentration on rate of freeze-drying and melting temperatures (T 0m ) were determined by differential
quality of the reconstituted products. In freeze dried scanning calorimetry (DSC) (Mettler TA 4000 system
yoghurt, poor texture and variable survival of yoghurt bac- equipped with TC11 TA Processor, DSC 30 measuring cell
teria have been reported (Rybka & Kailasapathy, 1997). and Graph Ware TA 72PS.2 software, Mettler, Greinfen-
In the present work, white skim yoghurt, plain or with see, Switzerland). Heat flow calibration was performed
added sugar and fruit (blueberries), were freeze-dried. Opti- with indium, temperature calibration was carried out with
mised freeze-drying parameters were extracted from the n-hexane, distilled water and indium. Aliquots of about
thermal analysis of the yoghurts, taking into account the 10 lg of samples were put into 40 ll hermetically-sealed
role of added ingredients. The distinct effects of freezing, aluminium DSC pans and scanned at 10 °C min1 under
freeze-drying, and storage were examined, with special 10 ml min1 dry nitrogen flow. Freezable solutions were
emphasis on the rheological properties, and on the lactic subjected to annealing before heating for the determination
bacteria viability of the reconstituted products. The effects of T 0g and T 0m . The glass transition temperatures were
of different rehydration ratios on texture were also reported as ‘‘onset’’ Tg. Determinations were made in trip-
considered. licate and each sample was scanned twice.

2. Materials and methods 2.4. Rheological analyses

2.1. Yoghurt samples The textural properties of fresh, frozen and reconsti-
tuted freeze-dried samples were determined by rheological
White plain skim yoghurt (Yomo, Milan, Italy) and fro- tests, by using a controlled stress rheometer (StressTech
zen blueberries (Boscobuono, Lazzati, Ponte, Pavia, Italy) rheometer, Reologica Instruments AB, Sweden), equipped
were purchased in a local supermarket. The following sam- with a cone-plate (4° cone angle, 40 mm diameter) measur-
ples were used: ing system. Before any measurements were taken, samples
‘rested’ for 20 min, allowing the stress induced during load-
(a) Plain yoghurt (y) (dry matter 11%, lactic acid 1.4%). ing to diminish. Dynamic tests were used to determine vis-
(b) Yoghurt mixed with 10% sucrose (Fluka, Steinheim, coelastic properties. The linear viscoelastic region was
Switzerland) (as an aqueous solution, 60% w/w) (ys). determined by stress sweeps at a constant frequency of
(c) Yoghurt mixed with 10% sucrose and 10% blueber- 1 Hz. Dynamic frequency sweeps were made with an
ries (previously pulped) (ysb). applied stress within the linear viscoelastic region, over a
0.3–10 Hz frequency ranges. All rheological determinations
were made at 4 °C ± 0.2 °C in triplicate.
2.2. Freezing and freeze-drying
2.5. Microbiological analyses
Samples of about 1 cm thickness were frozen on trays
and freeze-dried (Edwards Alto Vuoto, Mini Fast 1700, Microbiological analyses were performed on fresh, fro-
Milan, Italy) at a residual pressure of 20 Pa, corresponding zen and reconstituted freeze-dried samples. The fresh or
to the minimal vacuum chamber pressure reached by the frozen samples were mixed with sterile saline/peptone
apparatus, which allowed a sublimation temperature of water [8.5 g/l NaCl, 1 g/l bacteriological peptone (Oxoid,
34 °C. At the end of the drying process the chamber pres- Milan, Italy)], vortexed for 30 s and then serially diluted
sure was restored with dry nitrogen. Aliquots of about 10 g with the same diluent. The freeze-dried samples were mixed
404 E. Venir et al. / Journal of Food Engineering 80 (2007) 402–407

with sterile distilled water to the original volume. The Table 1

reconstituted samples were then serially diluted with sterile Tg dry, T 0g and T 0m of the yoghurt samples
saline/peptone water. Streptococcus thermophilus was Thermal parameters (°C) ya ys ysb
detected by surface plating onto duplicate plates of M17 Tg dry 48 ± 1 54 ± 2 54 ± 1
agar [M17 Broth (Oxoid) 37.25 g/l + Agar Bacteriological T 0g 57 ± 1 51 ± 1 52 ± 1
(Oxoid) 12.0 g/l]. Colony forming units (CFU) were T 0m 38 ± 1 34 ± 1 35 ± 1
counted after incubation at 37 °C for 48 h under microaer- a
See Fig. 1 for the legend.
ophilic conditions [Campygen (Oxoid)]. Typical colonies
(creamy and 1–1.5 mm in diameter) were randomly selected 1995) gave rise to an increase in both Tg dry and T 0m . Those
and confirmed by Gram-staining and catalase reaction. thermal parameters did not change significantly when blue-
Lactobacillus delbrueckii subsp. bulgaricus was detected berries were present. It should be noticed that any exposure
according to Rybka and Kailasapathy (1996), by surface- of freeze-drying material to temperatures above T 0m allows
plating onto duplicate plates of RCPB - pH 5 [Reinforced partial melting of the ice, and dilution and plasticization of
Clostridial medium (Oxoid) 38.0 g/l + Agar Bacteriologi- the concentrated amorphous phase. Such plasticization of
cal (Oxoid) 12.0 g/l + soluble Prussian blue 0.3 g/l (Fluka); the amorphous phase may result in loss of structure (col-
pH 5]. After incubation at 37 °C for 48 h under microare- lapse), reduced water removal rate, and poor product qual-
philic conditions [Campygen (Oxoid)] typical colonies ity (Bellows & King, 1973). Similar mechanisms relate the
(white centre, 1 mm diameter, surrounded by a wide clear Tg dry with the storage stability of the dry product, since
dark blue zone) were counted and randomly selected and storage temperatures above Tg dry may allow collapse
confirmed by gram-staining and catalase reaction. Analy- and reduction in stability of the dry material (Levine &
ses were repeated at least three times. Slade, 1988a, 1988b).
It was concluded that the addition of sucrose, with or
3. Results and discussion without blueberries, could improve the suitability of
yoghurt for both the freeze-drying and storage of the dry
3.1. Thermal properties product.

Fig. 1 shows the thermograms of anhydrous yoghurt 3.2. Rheological properties

samples. All the DSC traces showed a well resolved base
line shift, reversible at further rescans, consistent with a Dynamic tests on yoghurt indicated viscoelastic behav-
glass transition of the anhydrous materials (Tg dry). iour, within linear viscoelastic regions (at 4 °C) ranging
Anhydrous samples were reconstituted at a solid con- from 1 to 2.5 Pa, 0.8 to 2.8 Pa, and 0.09 to 0.3 Pa, respec-
centration of 30% (w/w) to determine the glass transition tively, for fresh, frozen/thawed and freeze-dried reconsti-
(T 0g ) and the initial melting (T 0m ) temperatures of the maxi- tuted samples. Mechanical spectra were determined at
mally freeze-concentrated solutions. Thermal parameters imposed stresses of 2, 1.5 and 0.2 Pa (corresponding strains
(T 0g , T 0m and Tg dry) of y, ys and ysb samples are reported were of 1.7% ± 0.29, 1.1% ± 0.15 and 0.6% ± 0.09), with
in Table 1. results shown in Fig. 2.
It is well known that product suitability for freeze-dry- Within the linear viscoelastic region, all samples exhib-
ing can be improved by increasing T 0g and T 0m through ited a weak-gel structure characterized by an elastic (G 0 )
changes in product formulations. Table 1 provides evi- modulus higher than the viscous (G00 ) one, with both moduli
dence that the addition of sucrose (Tg dry = 62 °C; Roos, slightly frequency-dependent (Ross-Murphy, 1995), and a
phase shift between 15° and 20°. Negligible destructuration
occurred following freezing and thawing (Fig. 2(B)). Con-
versely, a loss of overall consistency (Fig. 2(C)) marked
the freeze-dried reconstituted sample, indicating an irrevers-
ible weakening of the network structure, as also reported by
y Rybka and Kailasapathy (1997). It should be noted that a
Heat flow

contribution to the structural weakening of the reconsti-

ys tuted product could also derive from the mechanical energy
1 mW
ysb required for mixing with water, even when this is carried out
very carefully.
In an attempt to compensate for the reduction in
ENDO mechanical strength observed following freeze-drying and
reconstitution, freeze-dried samples were reconstituted to
20 30 40 50 60 70 80 90 100 100%, 70% and 60% of initial water content, corresponding
Temperature (°C)
to solid concentrations of 11%, 15% and 17%. It should
Fig. 1. DSC traces of anhydrous plain yoghurt (y); yoghurt + sucrose (ys) also be said that consistency and the reconstitution ratio
and yoghurt + sucrose + blueberries (ysb). Arrows indicate Tg (onset). are essentially a consumer choice.
 E. Venir et al. / Journal of Food Engineering 80 (2007) 402–407 405

G* G' G'' Viscosity* Phase could be attained at solids contents of 12–13 g 100 g1
(i.e., 70% of the initial moisture).
1000 90
A As shown in Table 2, the typical viscoelastic texture of
Viscosity* (Pa s)

Phase (degrees)
yoghurt was also retained in freeze-dried samples with
G*, G', G'' (Pa),

100 60 added sucrose and blueberries, reconstituted to their origi-

nal solid content. Effect of sucrose in maintaining the visco-
10 30 elastic properties may also be interpreted in terms of the
role of the hydroxyl groups in stabilizing the gel structure
(Bayarri, Duran, & Costell, 2004; Fu & Rao, 2001).
1 0
Freeze-dried yoghurt samples were stored at 4 °C up to
0.1 1 10
45 days. Samples were reconstituted at time intervals (70%
1000 90 of the initial water content) and the rheological properties
B are reported in Table 3. Data show that the structural
Viscosity* (Pa s)

Phase (degrees)
G*, G', G'' (Pa),

properties of all samples were maintained during storage.

100 60
3.3. Survival of lactic acid bacteria
10 30
The viable counts of S. thermophilus and L. delbrueckii
subsp. bulgaricus varied in each batch of fresh yoghurt,
1 0
0.1 1 10 with levels ranging from 104–108 CFU/g and 106–
107 CFU/g, respectively. For the survival studies, the
100 90 results of viability losses are shown as averages of log-unit
C reductions. Survival of lactic acid bacteria was evaluated in
Viscosity* (Pa s)

Phase (degrees)

both plain and sucrose–blueberry yoghurt samples after

G*, G', G'' (Pa),

60 freezing, freeze-drying and storage at normal refrigerator

10 temperature (4 °C).
30 In the plain yoghurt, freezing resistance appeared to be
similar for the two bacterial species, whereas, in the pres-
ence of sucrose and blueberries, L. delbrueckii subsp. bul-
1 0
0.1 1 10 garicus was significantly less susceptible (exhibiting a
Frequency (Hz) population reduction of 0.69 ± 0.44 Log units) than S.
thermophilus (exhibiting a population reduction of
Fig. 2. Mechanical spectra of yoghurt samples; plain yoghurt, A; frozen
1.69 ± 0.44 Log units) (Fig. 4). As a whole, the freeze-dry-
and thawed, B; freeze-dried and reconstituted (100% of initial water
content), C. ing process applied to the plain yoghurt, resulted in no sig-
nificant differences in terms of viability between L.
delbrueckii subsp. bulgaricus and S. thermophilus. The pres-
Fig. 3 shows G* (complex modulus), and g* (complex ence of sucrose and blueberries was found effective in sig-
viscosity) of plain yoghurt (control) and of freeze-dried nificantly increasing the survival of S. thermophilus cells,
samples reconstituted at different water contents. Mechan- with a population reduction of 1.52 ± 0.39 Log units for
ical properties close to those found for the plain yoghurt S. thermophilus against a reduction of 2.33 ± 0.83 Log
units for L. delbrueckii subsp. bulgaricus.
Storage at 4 °C for 45 days resulted in a small decrease
G* viscosity*
in lactic acid bacteria in freeze-dried plain products, with
the presence of sucrose and blueberries producing a protec-
1000 tive effect (data not shown).
G* (Pa); viscosity* (Pa s)

Bacterial resistance to freezing depends on a variety of

factors related to the microorganisms themselves and to
100
the manufacturing conditions (Fonseca, Beal, & Corrieu,
2001). The species influences bacterial resistance to freez-
10 ing: Streptococci generally survive better than lactobacilli,
due to differences in cell size and cell structure (Fonseca
et al., 2001). However, one of the most important condi-
1
tions affecting survival is the composition of the medium
9control
10 11 12 13 14 15 16 17 18 19 20 21
for freezing (Hubalek, 2003). In our study, the addition
Concentration of solids (gd.m. 100 g -1)
of sucrose and blueberries protected L. delbrueckii subsp.
Fig. 3. G* and complex viscosity* (at 1 Hz) of plain yoghurt (control) and bulgaricus, confirming the beneficial effect of these disac-
freeze-dried yoghurt (y) reconstituted to different solids contents. charides as a cryoprotective agent (Hubalek, 2003).
406 E. Venir et al. / Journal of Food Engineering 80 (2007) 402–407

Table 2
Dry matter, G*, G 0 , G00 , viscosity*, phase and G 0 /G00 ratio at 1 Hz of reconstituted (100% moisture) y, ys and ysb samples
Sample Dry matter (%) G* (Pa) G 0 (Pa) G00 (Pa) Viscosity* (Pa s) Phase (degrees) G 0 /G00
Freeze-dried (y) 11 37.1 ± 5.3 36.6 ± 6.8 10.6 ± 1.8 5.9 ± 0.8 16.0 ± 0.4 3.4
Freeze-dried (ys) 19.1 147 ± 9.6 140.7 ± 10.1 43.8 ± 2.7 23.5 ± 1.6 17.3 ± 0.4 3.2
Freeze-dried (ysb) 19.2 148.0 ± 41.0 140.0 ± 36.8 49.1 ± 16.9 23.6 ± 6.5 19.1 ± 1.5 2.8

Table 3
G*, G 0 , G00 , viscosity* and phase (1 Hz) of freeze-dried samples (y, ys and ysb) for different storage times at 4 °C
Samples Storage time (days) G* (Pa) G 0 (Pa) G00 (Pa) viscosity* (Pa s) Phase (degrees)
y 0 312 ± 90.9 298.2 ± 90.2 91.8 ± 15.8 49.7 ± 14.4 17.7 ± 2.4
15 245.7 ± 42.10 237.3 ± 41.7 64.6 ± 8.2 39.1 ± 6.7 15.3 ± 0.9
30 424.5 ± 52.6 412.0 ± 52.2 102.6 ± 8.3 67.6 ± 8.4 14.0 ± 0.7
45 356.0 ± 86.0 343.7 ± 83.5 92.0 ± 19.7 56.6 ± 13.7 15.1 ± 0.4
ys 0 n.d. n.d. n.d. n.d. n.d.
15 358.0 ± 46.2 343.0 ±45.3 103.9 ± 11.0 57.0 ± 7.4 16.9 ± 0.6
30 396.3 ± 21.5 380.0 ± 21.1 112.7 ± 3.5 63.1 ± 3.4 16.6 ± 0.4
45 356.0 ± 7.1 342.0 ± 8.5 100.6 ± 2.1 56.7 ± 1.1 16.4 ± 0.6
ysb 0 557.0 ± 77.3 533.0 ± 71.1 162.3 ± 31.0 88.7 ± 12.4 16.9 ± 1.2
15 533.8 ± 71.1 514.3 ± 66.1 142.8 ± 29.6 84.9 ± 11.1 15.4 ± 1.5
30 588.7 ± 29.5 566.0 ± 25.1 161.7 ± 22.7 93.7 ± 4.7 15.9 ± 1.6
45 622.0 ± 210.7 601.0 ± 2309.3 159.0 ± 33.9 98.3 ± 35.0 15.2 ± 2.1
Samples reconstituted to 70% initial moisture.

Log-unit reduction
4.00 presence of sucrose and blueberries. Furthermore, in the
S. thermophilus

L. delbrueckii

S. thermophilus

L. delbrueckii

presence of protectants, S. thermophilus survived better

S. thermophilus

3.50
S. thermophilus

than L. delbrueckii subsp. bulgaricus. As reported by

L. delbrueckii

3.00
Carvalho et al. (2004), the mechanisms underlying sucrose
L. delbrueckii

2.50
protection of dried cells could work by: (a) lowering the
2.00
phase transition temperature in dry membrane phospholip-
1.50 ids; (b) maintaining the dry proteins in their hydrated con-
1.00 formations; (c) protecting protein functionality, owing to
0.50 the formation of a glassy matrix during freeze-drying which
0.00 possesses high viscosity and low mobility. The high sur-
Frozen y Freeze-dried y Frozen ysb Freeze-dried ysb vival levels of S. thermophilus in a whey-sucrose protective
medium were potentially related to pH and the solid con-
Fig. 4. Loss of viability of Streptococcus thermophilus and Lactobacillus tent of the solution (Champagne & Gardner, 2001).
delbrueckii subsp. bulgaricus following freezing and freeze-drying with or
without sucrose and blueberries. The error bars show standard deviations.
Sucrose was found to decrease the dehydration inactivation
of L. plantarum (Linders, de Jong, Meerdink, & van’t Riet,
1997) and of Pantoca agglomerans (Costa, Usall, Teixido,
According to Panoff, Thammavongs, and Gueguen (2000) Garcia, & Vinas, 2000). When added to the drying medium
sucrose increased the survival rate of L. delbrueckii subsp. during storage of dried cells of L. bulgaricus, protection
bulgaricus by protection and by adaptation. was dependent on the type of sugar previously included
Damage to biological systems from freeze-drying can be in the growth medium (Carvalho et al., 2003).
attributed to two primary causes: changes in the physical
state of the membrane lipids and changes in the structure 4. Conclusions
of sensitive proteins (Leslie, Israeli, Lighthart, Crowe, &
Crowe, 1995). In general, the gram-positive cocci are the White skim yoghurt, plain or with added sucrose and
most resistant to freeze-drying, and Streptococcus is more blueberries, can be satisfactorily freeze-dried, provided that
resistant than Lactobacillus (Heckly, 1961). Similarly, thermal properties are taken into account for defining pro-
Wang, Yu, and Chou (2004) showed a higher percentage cess parameters. The addition of sucrose increased T 0m and
of survival after freeze-drying of S. thermophilus than Tg dry, thus improving the suitability of yoghurt for both
Lactobacillus acidophilus. In our study, although freeze- freeze-drying and storage. Yoghurt samples exhibited a
drying resulted in a higher percentage of survival in plain weak gel structure, which was not affected by freezing
yoghurt of S. thermophilus, differences were not significant. and thawing. By contrast, after drying and reconstitution
S. thermophilus survival was significantly improved in the a marked decrease in both the elastic and the viscous
 E. Venir et al. / Journal of Food Engineering 80 (2007) 402–407 407

moduli was observed, even if the ratio of the elastic to the Hubalek, Z. (2003). Protectants used in the cryopreservation of microor-
viscous modulus was maintained. The original mechanical ganisms. Cryobiology, 46, 205–229.
Kerwin, J., & Seddon, R. (2002). Eating in space – from an astronaut’s
strength was retained in yoghurt samples to which sugar perspective. Nutrition, 18, 921–925.
had previously been added or could be restored when plain Leslie, S. B., Israeli, E., Lighthart, B., Crowe, J. H., & Crowe, L. M.
samples were reconstituted at 70% of the initial moisture. (1995). Trehalose and sucrose protect both membranes and proteins in
The addition of sugar protected L. delbrueckii subsp. bul- intact bacteria during drying. Applied and Environmental Microbiology,
garicus from freezing and S. thermophilus from freeze-dry- 61, 3592–3597.
Levine, H., & Slade, L. (1988a). Principles of ‘‘cryostabilization’’
ing, reducing the level of bacterial mortality. technology from structure/property relationships of carbohydrate/
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concept for interpreting the creation and evaluation. London: Butter-
The excellent technical support of Eleonora Rizzotti and worths (pp. 149–180).
Linders, L. J. M., de Jong, G. I. W., Meerdink, G., & van’t Riet, K.
Andrea Tonizzo is gratefully acknowledged. This study (1997). Carbohydrates and dehydration inactivation of Lactobacillus
was supported by the ASI program: ‘‘Messa a punto di plantarum: The role of moisture distribution and water activity.
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