Lebensm.-Wiss. u.-Technol.

, 35, 171–176 (2002)

Growth Medium for Culturing Probiotic Bacteria for

Applications in Vegetarian Food Products
C. N. Heenan, M. C. Adams*, R. W. Hosken and G. H. Fleet

C. N. Heenan, M. C. Adams, R. W. Hosken: University of Newcastle, Centre for the Advancement of Food
Technology and Nutrition, P.O. Box 127, Ourimbah, NSW 2258 (Australia)
G. H. Fleet: University of New South Wales, Department of Food Science and Technology, Sydney,
NSW 2052 (Australia)
(Received April 27, 2001; accepted August 20, 2001)

Vegetarian probiotic foods by definition, must be free from all animal-derived ingredients. This not only includes the product
ingredients but the probiotic inoculum as well. Probiotic starter cultures are traditionally grown and stored in media containing milk
or meat-derived ingredients. The presence of these ingredients makes the probiotic cell concentrates unsuitable for use in
vegetarian products and thus creates the need for a growth medium free from animal-derived ingredients. This study investigated the
growth of two strains of Lactobacillus acidophilus (MJLA1 and La-5), L. paracasei ssp. paracasei (LCSH1 and 01) and
Bifidobacterium lactis (BDBB2 and Bb-12) in five media. Growth parameters such as maximum cell concentrations, lag phase and
growth rate, were calculated using the growth curves generated by DModel, Institute of Food Research, U.K. Results showed that
several media were suitable, with results comparable or better than the commonly used laboratory media MRS and RCM. Media
containing 25 g/L soy peptone, yeast extract and glucose monohydrate gave the most desirable results with the strains examined.

r 2002 Elsevier Science Ltd. All rights reserved.

Keywords: probiotics; lactic acid bacteria; vegetarian; acidophilus

Introduction not share the good growth characteristics of the

Increase in demand for functional foods throughout the
world, has led to the development of more specialised
products. Probiotic foods contain viable bacteria that
beneficially influence health and nutrition when con-
sumed. Some of the proposed health benefits provided
by probiotic species include an aid to digestion;
reduction in episodes of diarrhoea; prevention and
suppression of colon cancer; lowering of cholesterol
and stimulation of the immune system (Gilliland et al.,
1985; Marteau & Rambaud, 1993; Sanders, 1994;
Perdigon et al., 1995; Kaila & Isolauri, 1996).
Lactic acid bacteria (LAB), the most commonly used
probiotic group, are fastidious organisms and in the
laboratory, are typically grown on complex media such
as MRS (de Man et al., 1960) or Reinforced Clostridial
Medium RCM (Hirsch & Grinsted, 1954) that include
meat or milk extracts. LAB are produced for commer-
cial use as starter culture concentrates for the manu-
facture of fermented foods and beverages or for
incorporation into unfermented food products at the
time of manufacture. Many strains of probiotic LAB do
*To whom correspondence should be addressed. Tel.: 61 2 4348 4135;
fax: 61 2 4348 4145; E-mail: madams@mail.newcastle.edu.au
0023-6438/02/$35.00
r 2002 Elsevier Science Ltd. All rights reserved.
usual industrially exploited strains like Lactobacillus
delbrueckii ssp. bulgaricus and Streptococcus salivarius
ssp. thermophilus, and special attention is required to
achieve high cell concentrations.
The choice of growth medium for producing commercial
quantities of probiotic bacteria is constrained by costs,
efficiency of cell production and ease of harvest
(Champagne et al., 1991). When producing bacteria on
a large scale, maximum cell concentration and the time
taken to reach this level have a significant impact on
cost. Commercially, competitive growth medium should
comprise inexpensive ingredients and permit bacteria to
reach a high cell density in a short time frame to reduce
production costs. The ingredients need to be in solution
or be able to be solublised, to allow the effective harvest
of the bacterial cells by either filtration or centrifugation
(Mitchell & Gilliland, 1983).
Specific formulations for industrial growth media are
rarely reported to maintain the commercial advantage of
the company marketing the strains. In general, com-
mercial bacterial cell production has used milk-based
media and more recently whey-based media, a bypro-
duct from cheese manufacture (Gilliland, 1985a). Milk is
nutritious containing carbohydrates, fat, casein protein,
vitamins and minerals (Marshall & Tamime, 1997).
doi:10.1006/fstl.2001.0833
All articles available online at http://www.idealibrary.com on

171
lwt/vol. 35 (2002) No. 2
Some probiotic strains do not grow well in milk or whey
(Ahmed et al., 1990; Osborne, 1992; Klaver et al., 1993;
Saxena et al., 1994) as the isolates lack sufficient
proteolytic ability to obtain nitrogen from the milk or
whey protein (Gilliland, 1985b; Champagne et al., 1996;
Shihata & Shah, 2000). If the sterilising heat treatment is
not adequately controlled, protein can form a precipi-
tate and consequently compromise bacterial cell harvest-
ing (Marshall, et al., 1982; Mitchell & Gilliland, 1983;
Modler & Villa-Garcia, 1993).
With consumer vegetarianism increasing in many parts
of the world (Beardsworth & Keil, 1991; 1992; Anon.,
1994) food manufacturing companies now have to
market appropriate vegetarian probiotic products in
addition to the traditional fermented milk. Vegetarian
probiotic foods need to be altogether free from animal-
derived ingredients to be classified as vegetarian.
Consequently, for inclusion in vegetarian foods, the
probiotic inoculum should also be cultured in a growth
medium free of animal-derived ingredients to prevent a
carry-over of these ingredients into the product.
Ideally, the medium should produce cell concentrations
around 109 cfu/mL and above. Cell concentrations
around 1010 cfu/mL have been achieved with continuous
fermentation and external pH control (Misra & Kulia,
1991; Modler & Villa-Garcia, 1993). When cell concen-
tration is calculated as dry weight, a good harvest
equates to between 1.0 and 2.0 mg/mL depending on the
species examined (Cogan et al., 1971; Barreto et al.,
1991; Coventry & Hickey, 1991).
This investigation is aimed at developing a growth
medium, free from animal-derived ingredients, with
potential for commercial cultivation of probiotic strains
for application in vegetarian foods. It reports several
soy-based growth media supplemented with yeast
extract, sugars and internal buffering.
Materials and Methods
Probiotic bacterial strains
Commercial probiotic strains of Lactobacillus acidophi-
lus, L. paracasei ssp. paracasei and Bifidobacterium lactis
were obtained from Christian Hansen (Bayswater,
Australia) and Gist Brocades (Moorebank, Australia)
as shown in Table 1.
Media composition and preparation
The composition of the media used for culturing LAB is
shown in Table 2. Final recipes for each medium
composition were derived from a series of preliminary
experiments using soy protein, soy peptone, yeast,
malt and vegetable extracts and various sugars. The
preliminary experiments consisted of initial and final
cell concentrations and were conducted without
replication.
Medium ingredients used in the comprehensive growth
curves were dissolved in distilled water and the pH
adjusted with 1 mol/L NaOH or HCl to the values
indicated. For lactobacilli, 75 mL of growth medium
172
Table 1 Sources and strains of probiotic bacteria
Strain Source Code
Lactobacillus acidophilus La-5 Chr. Hansen La-5
L. paracasei ssp. paracasei 01 Lp-01
Bifidobacterium lactis Bb-12 Bb-12
L. acidophilus MJLA1 Gist Brocades MJLA1
L. paracasei ssp. paracasei LCSH1 LCSH1
B. lactis BDBB2 BDBB2
Table 2 Composition of media used for the growth of
probiotic organisms
Medium Ingredients pH
MRS Oxoid (CM 359) 7.0
RCM Oxoid (CM 149) 6.8
SPY 1 10 g/L soy peptone, 10 g/L glucose 7.0
monohydrate, 10 g/L yeast extract
SPY 2 25 g/L soy peptone, 25 g/L glucose 7.0
monohydrate, 25 g/L yeast extract
SPY 3 25 g/L soy peptone, 50 g/L glucose 7.0
monohydrate, 25 g/L yeast extract
SPY 4 SPY 2 plus 15.1 g/L KH2PO4, 3.4 g/L 6.3
Na2HPO4
SPY 5 SPY 2 plus 25 g/L inulin (Raftilines, Orafti) 6.7
was dispensed into culture bottles, sealed with a rubber
septum and aluminium crimp cap, then autoclaved
(15 min, 121 1C). The medium was then flushed with
CO2 gas, for 30 min and tempered to 37 1C before use.
Media used for bifidobacteria were dispensed into
culture bottles, heated to 100 1C for 15 min to eliminate
O2, sealed, flushed with N2 for 20 min and then
autoclaved (15 min, 121 1C). The medium was flushed
with N2 for 20 min and tempered to 37 1C before use.
Comprehensive growth trials
Lactobacilli and bifidobacteria were cultured in either
RCM or MRS medium respectively, for 24 h at 37 1C,
followed by subculture into the same media for a further
15 h. Growth media were inoculated at an approximate
concentration of 104 cfu/mL for lactobacilli and
105 cfu/mL for bifidobacteria. Lactobacillus spp. were
grown in MRS, SPY 1, SPY 2, SPY 3 and SPY 4. In
addition to these media, Bifidobacteria spp. were also
grown in RCM and SPY 5.
Cultures were incubated for 36 h at 37 1C on a shaking
incubator (100 rev/min) for lactobacilli and unshaken
for bifidobacteria and sampled regularly for the
determination of cell concentrations and pH. Viable
cell concentrations were measured by spread plate
counting in duplicate on MRS or RCA agar. MRS
plates were incubated in 80 mL/L CO2 incubator
(Napco, 6200) and RCA plates in anaerobic jars
(Anaerogen, Oxoid Heidelberg, Australia) at 37 1C for
48 h. Resulting colonies were counted and cell concen-
trations were calculated. Growth curves were con-
structed using Dmodel (Baranyi & Roberts, 1995) and
 lwt/vol. 35 (2002) No. 2

were used to calculate growth rates, lag phase, popula-
tion maximum and the time taken to reach population
maximum. Each experiment was conducted three times
and analysed by 1- and 2-factor ANOVA using the
computer program Statgraphicss (Version 2.0, Stat-
graphics Corp.). Due to the experimental design, the
Lactobacillus and Bifidobacterium strains were analysed
as separate groups.
Dry biomass determination
Bacterial dry biomass was determined for all strains
grown in MRS or RCA, and SPY 2. Strains were grown
to early stationary phase and harvested by centrifuge.
Twenty millilitres of culture was centrifuged (5000  g)
in pre-weighed centrifuge tubes, washed with 5 mL
1.0 g/L peptone and then the pellet was dried for 20 h
at 60 1C. Pellets were equilibrated 0.1aw by exposure to
saturated lithium chloride for 96 h in a sealed container.
Centrifuge tubes were reweighed and the weight of
biomass calculated. Viable cell counts of the early
stationary phase cultures were conducted by spread
plating immediately prior to cell harvest.
Results and Discussion
Preliminary growth trials
Preliminary growth trials (data not shown) revealed that
soy protein (Isolate 760, Protein Technologies Interna-
tional, U.S.A.) was not a suitable ingredient as it did not
dissolve. The precipitate persisted even after treatment
with pepsin hydrolysis (Mitchell & Gilliland, 1983). This
ingredient was replaced with soy peptone made by
papyric hydrolysis (Oxoid). Using the revised formula,
cell concentrations equal or greater than MRS were
achieved with several media. There was some variation
depending on the organism, with no one medium
proving suitable for all of the probiotic strains. Probiotic
isolates grew best in a basal medium of soy peptone,
yeast extract and glucose monohydrate. When the sugar
concentration was doubled to 100 g/L, there was no
equivalent increase in cell concentration, indicating that
sugar is not a limiting factor for the organisms. The
extra sugar was substantially detrimental to the growth
of L. acidophilus MJLA1, B. lactis BDBB2, B. lactis
Bb-12 strains and of no benefit to the other strains.
Comprehensive growth trials
Maximum cell populations (Table 3) were dependent on
the strain and culture medium (Po0.05). The two
strains of L. acidophilus gave the highest concentration
on MRS (Po0.05), although the cell populations of
L. acidophilus MJLA1 in SPY 2, SPY 3 and SPY 4 were
only marginally lower than MRS. The two strains of
L. paracasei ssp. paracasei showed no significant
differences in maximum populations obtained on SPY
2, SPY 3, SPY 4 and MRS (Table 3). Bifidobacterium
strain BDBB2 and Bb-12 achieved cell populations
equal to MRS in SPY 2 and SPY 3 (Po0.05). SPY 1
medium was inferior to the other media, producing the
lowest maximum population in L. acidophilus MJLA1,
L. acidophilus La-5, L. paracasei ssp. paracasei LCSH1
and B. lactis Bb-12 (Po0.05).
Lactobacillus acidophilus La-5 grew poorly in all of the
soy-based media, with reduced growth rates and a short
exponential phase. Lactobacillus acidophilus La-5 may
not be able to utilise the soy-based media or there may
be another factor limiting the growth. Lactobacillus
acidophilus La-5 exhibited rough colony morphology
unlike the other Lactobacillus strains used in this study.
Klaenhammer and Kleeman (1981) also detected re-
duced growth rates, bile tolerance and hardiness in
rough colony variants of L. acidophilus.
No growth benefit was gained by incorporating the
prebiotic inulin into the growth medium for bifidobac-
teria (SPY 5). Inulin is a plant-derived fructo-oligosac-
charide (Raftilines Orafti, Tienen, Belgium), that is
thought to be selectively utilised by Bifidobacterium
species in the human gut (Roberfroid, 1993). The
maximum populations obtained from both Bifidobacter-
ium strains in SPY 5 did not exceed the maximum
populations from the same medium without inulin (SPY
2), and SPY 5 was detrimental to strain B. lactis Bb-12.
The results indicate that the Bifidobacterium strains
preferentially use glucose over inulin under these
conditions. In the gastrointestinal tract, glucose would
not normally be available to microflora, having already
been absorbed by the host. The positive response of
strain B. lactis BDBB2, as opposed to the negative
response of B. lactis Bb-12 in inulin, indicates that inulin

Table 3 Maximum cell concentrations (log10 cfu/mL) of probiotic bacteria in different media
Strain
MJLA1 La-5 LCSH1 Lp-01 BDBB2 Bb-12
Medium Mean SD Mean SD Mean SD Mean SD Mean SD Mean SD
MRS 9.13* 70.06 78.73* 70.03 8.86* 70.18 9.20 70.07 8.24* 70.21 8.61* 70.36
SPY 1 8.34 70.03 5.44 70.56 8.34 70.33 9.11 70.54 8.38* 70.06 7.40 70.10
SPY 2 8.86 70.07 6.94 70.03 8.95* 70.09 9.26 70.06 8.51* 70.18 8.11* 70.36
SPY 3 8.99 70.02 7.02 70.03 8.83* 70.10 9.23 70.08 7.99 70.58 7.99 70.50
SPY 4 8.84 70.02 7.14 70.06 8.95* 70.13 9.14 70.09 7.77 70.56 7.67 70.61
SPY 5 8.10 70.12 6.60 70.03
RCA 6.30 70.13 6.56 70.02
*Significantly the highest maximum cell concentration produced by that strain (Po0.05), SD=standard deviation (n=3)

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Table 4 Time (hours) taken to reach maximum cell concentration for probiotic bacteria in different media
Strain
MJLA1 La-5 LCSH1 Lp-01 BDBB2 Bb-12
Medium Mean SD Mean SD Mean SD Mean SD Mean SD Mean SD
MRS 19.0 70.00 24.0 71.73 23.3 71.15 28.7 71.53 17.3 73.51 16.0 71.00
SPY 1 17.7 70.58 13.3 72.89 23.3 70.58 26.0 72.65 15.0 71.73 13.0 73.46
SPY 2 18.3 71.53 16.7 70.58 26.3 70.58 28.0 71.00 14.3 70.58 13.7 72.31
SPY 3 19.0 71.00 17.0 71.00 24.0 71.00 28.0 70.00 13.7 73.79 13.0 72.65
SPY 4 18.3 70.58 15.0 70.00 24.0 71.00 29.7 70.58 15.0 74.36 14.0 75.29
SPY 5 22.3 73.21 12.7 72.31
RCA 14.7 71.15 15.7 70.58
SD=standard deviation, (n=3)

would probably be of greater benefit to B. lactis BDBB2
in vivo.
The average time taken for the Lactobacillus strains to
reach stationary phase, although significantly different
between strains and media, was not practically signifi-
cant (Table 4). Each strain exhibited a small range
of growth times across all media, indicating a strain-
specific, rather than medium-specific response. Time
taken for the Bifidobacterium strains to reach maximum
population was independent of medium (Po0.05). Both
the Bifidobacterium strains reacted similarly in all the
media except SPY 5 where B. lactis BDBB2 took nearly
10 h longer than B. lactis Bb-12 to reach maximum
population. This is reflected in B. lactis BDBB2 on
reaching a much higher cell concentration than B. lactis
Bb-12 in this medium. All strains except L. paracasei
ssp. paracasei LCSH1 reached stationary phase faster in
SPY 2 than MRS, although the difference was only
significant for B. lactis BDBB2 and L. acidophilus La-5
(Po0.05).
There was no significant difference between lag phase
attributable to the growth media. Lag phase was a
particular feature of each Lactobacillus isolate
(Po0.05), with L. paracasei ssp. paracasei LCSH1
having the longest average lag phase of 3.7 h, followed
by L. acidophilus MJLA1 and then L. paracasei ssp.
paracasei Lp-01 and L. acidophilus La-5 equally.
Interestingly, L. paracasei ssp. paracasei LCSH1 with a
exception of SPY 1 and SPY 2, the pH of the other
media did not substantially alter until after stationary
phase, indicating that pH did not limit Bifidobacterium
growth. Both Bifidobacterium strains had an extremely
short stationary phase in SPY 4 medium and entered
death phase after only 15 h (Fig. 1). The pH remained
above 5.5, excluding a change in pH as the cause of the
onset of the death phase. A possible accumulation of
metabolites produced in the SPY 4 may have caused cell
death. This is in agreement with Herrero (1983) who
stated that anaerobic fermentation end products limit
growth.
Dry biomass
Dry biomass, cell concentrations per millilitre and
individual cell mass produced in MRS and SPY 2
growth medium were determined for all strains
(Table 5). Significantly, different bacterial biomass was
9
8
Cell concentration (log10 cfu/mL)

long lag phase still reached stationary phase faster than

the strain L. paracasei ssp. paracasei Lp-01, due to faster 7
growth rates. Neither of the Bifidobacterium strains
exhibited a calculable lag phase.
6
Change in media pH during incubation (data not
shown) reflected how vigorously each strain grew in
that medium. The more vigorous the growth, the lower
5
the final pH. The buffering system used in the SPY 4
medium only delayed by 2–6h the drop in pH,
depending on the Lactobacillus strain. The small 4
difference between maximum lactobacillus populations
achieved in SPY 4 and SPY 2, the same medium without
buffering, indicates that either pH was not the over- 3
whelming factor limiting growth or the delay in pH 0 10 20 30
change was not sufficient to produce increased cell Time (h)
concentration. Fig. 1 Cell concentration of Bifidobacterium lactis strain
The Bifidobacterium strains did not acidify the media to BDBB2 (triangles) and Bb-12 (circles) in SPY 2 (solid) and
the same degree as the Lactobacillus species. With the SPY 4 (hollow)

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Table 5 Biomass and viable counts for probiotic bacteria in MRS and SPY 2 media.
PY 2 MRS cell SPY 2 cell
MRS biomassw biomassw concentration* concentration* Cell weight in Cell weight in SPY
(mg/mL) (mg/mL) (log10 cfu/mL) (log10 cfu/mL) MRS (pg) 2 (pg)
Strain Mean SD Mean SD Mean SD Mean SD Mean SD Mean SD
MJLA 1 1.98b
70.03 2.74a
70.04 8.95c
70.02 8.60d
70.09 2.22 f
70.07 6.91 e
71.40
La-5 1.07a 70.03 0.07b 70.04 8.51c 70.05 7.11d 70.09 3.32 70.34 5.33 71.81
BDBB2 0.85b 70.06 1.32a 70.06 8.83 70.05 8.80 70.04 1.25 f 70.07 2.08e 70.15
Bb-12 0.67b 70.08 1.20a 70.03 8.79 70.09 8.73 70.03 1.09 f 70.15 2.25e 70.19
LCSH1 2.19a 70.11 0.92b 70.11 9.13c 70.02 8.68d 70.04 1.62 f 70.02 1.92e 70.06
Lp-01 2.28a 70.01 1.35b 70.08 9.12c 70.05 8.71d 70.08 1.74f 70.20 2.66e 70.31
a,b
Indicates a significant difference in biomass obtained in MRS and SPY 2 media for that strain (Po0.05).
c,d
Indicates a significant difference in cell concentration obtained in MRS and SPY 2 media for that strain (Po0.05).
e,f
Indicates a significant difference in cell weight from MRS and SPY 2 media for that strain (Po0.05).
SD=standard deviation; waverage biomass (n=6); *average counts (n=3).

produced in MRS and SPY 2 for all strains. Lactoba-
cillus acidophilus MJLA1, B. lactis BDBB2 and B. lactis
Bb-12 had greater biomass per millilitre in SPY 2 than in
MRS (Po0.05), despite there being no significant
difference in viable counts for both Bifidobacterium
strains, and L. acidophilus MJLA1 having a significantly
greater cell concentration in MRS.
All strains grown in SPY 2 had a greater individual cell
mass than the same strain grown in MRS (Po0.05),
although the L. acidophilus La-5 result was not
significant. Both strains of L. acidophilus had cell
weights much greater than the other species under
examination in both media. Porubcan and Sellars (1979)
demonstrated that the turbidity generated by the L.
acidophilus (ATCC 4356) in MRS was not solely due to
bacterial cells and speculated that the initially soluble
medium formed a precipitate when exposed to L.
acidophilus metabolic products. A similar phenomenon
may occur with a precipitate adding to the dry weight
per millilitre, creating a higher cell weight than the other
species examined.
Lactobacilli cell morphology is affected by media
composition, such as alterations in rod length caused
by the availability of oleic acid esters (Kandler & Weiss,
1986). Although L. acidophilus bacilli are longer than
L. paracasei ssp. paracasei and bifidobacteria cells
(microscopic observation), when the dry cell mass is
calculated using a model of 1 mm in width, increasing cell
length and a dry solids ratio of 15% (data not shown),
the differences in the L. acidophilus results to the
other species are too great to be solely attributed to
cell length.
With L. paracasei ssp. paracasei LCSH1 and
L. paracasei ssp. paracasei Lp-01, greater cell weights
were obtained from SPY 2 (Po0.05) but there was
significantly higher biomass per millilitre produced in
MRS (Po0.05). The greater cell concentrations pro-
duced in MRS account for the higher biomass, as
although SPY 2 cells weigh more, there were less of them
present in SPY 2 medium. When there was no significant
difference with cell concentrations, as with bifidobacter-
ia strains BDBB2 and Bb-12, the greater weight per cell
in SPY 2 also produced greater biomass per millilitre
growth medium (Po0.05).
For the production of probiotic starter concentrates,
actual biomass is not as relevant as viable counts. In
food applications, the viability of probiotic strains, cell
concentration in the product, is used as the measure for
probiotic suitability, rather than the probiotic biomass
added to the product. Additionally, biomass does not
take into account viability or sublethally damaged cells
and may be misleading by including the weight of dead
cells. Furthermore, as in this research, cells can weigh
significantly more when grown in one medium com-
pared to another, generating more biomass with less
viable cells actually present.
Conclusions
Several of the growth media containing no animal-
derived ingredients, were suitable for culturing probiotic
isolates destined for vegetarian food products. Bacterial
growth in SPY 2, SPY 3 and SPY 4 was similar or better
than in MRS. SPY 2 has the greatest potential as an
industrial growth medium for probiotics, justifying
further examination for optimisation for commercial
production. The higher sugar concentration in SPY 3 or
buffering in SPY 4 did not produce a sufficiently
substantial increase in probiotic cell concentration to
warrant the expense of extra ingredients. Furthermore,
there were disadvantages in using SPY 4 media to grow
bifidobacteria, as the strains entered decline phase after
only 16 h. Effective media buffering is desirable to
prevent sublethal injury caused by exposure to the low
pH during incubation (Gilliland, 1985a). Alternative
buffering methods such as automated external buffering
systems, would be worthwhile examining when optimis-
ing SPY 2 for scaled-up probiotic cell production. This
research, as well as identifying suitable vegetarian
growth media, also highlighted the necessity of selecting
the right strains for each application. The phenotypic
properties of each of the probiotic strains have a
significant impact on the performance of the strain in
any growth medium. Growth time to stationary phase,
lag phase and general growth vigour were features of
each strain rather than the test medium. New probiotic

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lwt/vol. 35 (2002) No. 2
strains must be examined to identify industrially
robust isolates to avoid strains such as L. acidophilus
La-5, that are not suited to vegetarian growth media.
With the development of a suitable growth medium
containing no animal-derived ingredients, vegetarian
probiotic products can now be manufactured to a
quality that is acceptable to the most discerning
vegetarians.
Acknowledgements
Financial and technical support from the University of
Newcastle, Australian Research Council and Sanitarium
Health Food Company, Australia, and the provision of
probiotic cultures from Christian Hansen and Gist
Brocades are gratefully acknowledged.
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