Technological & Commercial Applications of Lactic Acid Bacteria;

Health & Nutritional Benefits in Dairy Products

Stanley E. Gilliland, PhD
Department of Animal Science
Food & Agricultural Products Center
Oklahoma State University
Stillwater, OK 74078
USA
Key words: Lactobacillus, Bifidobacterium, milk, probiotics, storage stability, nutritional benefits, health benefits
Introduction
Probiotics have been defined as selected viable microorganisms used as dietary supplements having
potential for improving health or nutrition of man or animal following ingestion. The primary probiotic bacteria
associated with dairy products have been Lactobacillus acidophilus, Lactobacillus casei and bifidobacteria. More
recent taxonomic studies involving sophisticated techniques have resulted in reclassification within some of the
species of Lactobacillus listed as probiotics to the Lactobacillus acidophilus group and the L. casei group (Table 1).

T a ble 1 . Prob iotic ba cte ria th a t m ay b e associated w ith m ilk p rod ucts
L a c to b a c illu s a c id o p h ilu s group
L a ctobacillus reuteri
L. a cid op h ilu s
L. am ylovorus
L a ctobacillus plan ta ru m
L. crispatus
B ifidobacterium sp ecies:
L. g a sseri
L. jo h n so n n i
la c tis
L a ctobac illu s c a sei group
longum
L. ca sei
adole scentis
L. paracasei
anim a lis
L. rh a m n o su s

b ifid u m
breve
infa ntis

Other lactobacilli include L. reuteri and L. plantarum. Bifidobacterium species include longum, adolescentis,

animalis, bifidum, breve infantis, and lactis (Holzapfel et al 1998, Klein et al 1998, Reid 1999). Some of them or
closely related organisms have been used for centuries in the manufacture of cultured dairy products. For this
reason, they are generally regarded as safe. Which means there is minimal concern to their being used as dietary
adjuncts in dairy products.
Milk or milk products provide an excellent carrier for these probiotic organisms. Most of them can readily
utilize lactose as an energy source for growth. Thus, an important requirement for the growth in the intestinal tract

is provided by the milk. Milk proteins also provide important protection to the probiotic bacteria during passage
through the stomach (Charteris et al 1998). Compared to cells suspended in buffered saline all cultures tested
survived exposure to simulated gastric juice much better when milk protein was present.
While traditional starter cultures used in the dairy industry are selected for their ability to rapidly produce
desirable organoleptic qualities of cultured dairy products, the probiotic bacteria should be selected for the potential
to provide specific health or nutritional benefits following consumption. Unfortunately, in many cases the probiotic
bacteria included in the cultured dairy products have not been selected for specific functions, but rather are added
just so the manufacturer can say the organism is there. Considerable variation occurs among strains of these bacteria
with regard to their potential for producing health and nutritional benefits. Thus, proper selection of the strain to be
included as a probiotic becomes important.
A number of dairy products are marketed as containing probiotic bacteria. However, the most widely
encountered one is yogurt. It is not uncommon today to read on the label of yogurt products that the product was
made with a culture including L. acidophilus or bifidobacteria species. These two cultures are not ones that have
traditionally been used to manufacture yogurt.

The traditional yogurt starter cultures of course include

Streptococcus salivarius ssp thermophilus and Lactobacillus delbrueckii ssp bulgaricus. In the United States, we
also have what is referred to as nonfermented acidophilus milk in which cells of L. acidophilus are added to freshly
pasteurized milk. The milk is then stored under refrigeration so the probiotic bacteria does not grow in the milk, but
rather the milk serves as the carrier for the organism. There are many other fermented products in the world with a
dairy base, which contain probiotic bacteria. An example is Yakult, which is made with a selected culture of

Lactobacillus casei. Other that health food store and pharmaceutical products few if any dried milk products
containing probiotics are available.

Potential benefits

There are a number of potential benefits that might be derived from consuming dairy products containing
probiotic bacteria (Table 2). In this presentation, I will not attempt to discuss in detail all of the potential benefits
that might be derived from the consumption of dairy products containing probiotic bacteria.

Table 2. Potential benefits from probiotics

* Control of intestinal infections

* Stimulation or modulation of the immune system
* Improve lactose (or other nutirent) utilization
* Control of some cancer
* Control of serum cholesterol levels

Since the time of Eli Metchnikoff (1904) much has been learned about microbiology and many reports
have been published concerning the benefits of lactobacilli in relation to the intestinal tract. The interest in the role
of lactobacilli and other lactic acid bacteria in the microecology of the intestinal tract was regenerated in the last
twenty years. Unfortunately, many of the early studies on the use of lactobacilli to control intestinal infections were
not well designed. In most cases, the lactobacilli were used as a therapeutic rather than a preventative agent and in
many cases adequate controls were not included in the studies. Furthermore, in many of these studies, there was
very little information concerning the particular culture of lactobacilli involved, especially with regard to its origin
or its potential for producing the desired effect.
It may be more reasonable to consider probiotics as a prophylactic rather than a therapeutic treatment for
intestinal infections.

One of the studies which confirms the advantage of considering L. acidophilus as a

prophylactic treatment for controlling intestinal infections was done by Watkins and Miller (1983). In their studies
they used germ-free chickens as the animal model. In the prophylactic experiment, the chickens were fed a culture
of L. acidophilus initially. Following two days, the chicks were divided into three groups. One group served as a
control. The other two groups were challenged with either Salmonella typhimurium or Staphylococcus aureus. One
group for each pathogen received no further treatment, and the third group was additionally fed L. acidophilus at
two-day intervals for ten days.

In the therapeutic experiment, germ-free chickens were fed the pathogenic

organisms initially, then divided into three groups. One group served as control; one group was fed L. acidophilus
on day two; and the third group received L. acidophilus at two-day intervals for a total of five treatments following
inoculation with the pathogen. In the therapeutic experiments, L. acidophilus had minimal affect. Results indicated
that the animals fed L. acidophilus initially (i.e. prior to the challenge with the pathogen) survived much better than
did those that were challenged with the pathogen first. Furthermore, the continued feeding of L. acidophilus
following the challenge of the chickens with the pathogens was the best form of treatment. These results suggest it

is important to have L. acidophilus initially and to provide the organism on a continuing basis in the diet for best
control of these pathogens.
In the dairy industry, we have known for many years that there was variation among strains and species of
starter culture bacteria with regard to their ability to produce the desired changes in the milk being fermented. Thus,
it should not be surprising that there would be variation among strains and species of probiotic microorganisms with
regard to their ability to produce inhibitory action toward pathogenic microorganisms. Furthermore, we should not
expect one strain or species of probiotic microorganisms to provide all of the potential benefits that might be
possible from consumption of these organisms.
Feeding trials are difficult to conduct and very expensive with regard to showing the influence of probiotic
organisms on intestinal pathogens. Thus, some sort of laboratory screening of the cultures prior to their use as
probiotics seems desirable. As an example, we have conducted studies recently comparing six strains of L.

acidophilus of bovine origin for the ability to inhibit Escherichia coli 0157:H7 in associative cultures in laboratory
media. For these experiments, two strains of the pathogenic E. coli were included. Broths were inoculated with
each at approximately 3x104 colony forming units (CFU)/ml. The inoculated broths were divided into seven parts.
One for each strain of E. coli served as control and each of the other portions was inoculated with different strains of

L. acidophilus (1x105/ml). All tubes were incubated six hours at 37C after which the numbers of E. coli were
enumerated on Violet Red Bile Agar. The results revealed variation among the strains of L. acidophilus with regard
to the percentage inhibition of the pathogen. Four of the strains were significantly more inhibitory than the other
two. Thus, if we were to select a strain of L. acidophilus as a probiotic for helping control E. coli 0157:H7 in cattle,
we would want to select one of these which exhibited the greatest amount of inhibition in the laboratory tests.
Similar variations should be expected when selecting strains of a probiotic to control intestinal pathogens in humans.
Various probiotic bacteria have been shown in scientific studies to aid in control of intestinal pathogens in
humans. Examples of these are listed in Table 3. The studies listed in this table focus on control of diarrhea in
children through the use of probiotics.

Table 3. Examples of published studies reporting use of probiotic bacteria to exert control of intestinal
infections in humans.
Benefit
Reference
Significant reduction in duration ot diarrhea in
Gozalez et al. 1995
children in Argentina. Milk formulated with L.
casei & L. acidophilus
Beneficial effect in control of rotavirus diarrhea
in children by L. reuteri .

Shornekova et al. 1996

Reduced severity of diarrhea in children by L.

casei.

Pedone et al. 1999

Control of viral diarrhea in children through use

of Lactobacillus & Bifidobacterium

McNaught & MacFie. 2001

There have been several possible mechanisms proposed to explain how probiotic bacteria can inhibit
pathogenic microorganisms in the intestinal tract. These include the production of antimicrobial agents, competitive
exclusion, competition for nutrients, and stimulation of the immune system. Of these possible mechanisms, the one
which seems to be receiving the most attention today is stimulation or modulation of the immune system (Alvarez et
al, 1998, Perdigon & Alvarez, 1992, Perdigon et al, 1995). In this groups studies, the consumption of L. casei
caused the body to secrete antimicrobial substances into the intestine. Bifidobacteria fed to mice also caused similar
results in another study (Fukushima et al, 1999). This mechanism appears to have a lot of potential as far as
answering the questions as to how these bacteria might exert inhibitory action toward intestinal pathogens. While
most of the work on modulation of the immune system has involved animal studies, there have been studies focusing
on the effect of feeding cells of L. acidophilus on the immune system of humans. In one study (Link-Amster et al,
1994) an increase in the secretion into the intestine was observed of substances which were considered inhibitory for
certain intestinal pathogens.

Currently the final answer regarding the most likely mechanism for controlling

intestinal pathogens by probiotic bacteria is not clear. Certainly additional research is needed. Ideally this will
involve challenge studies using human subjects, but it is difficult to get people to participate in such a study and to
get approval by university research review committees.
Modulation of the immune system through the use of probiotics also has been shown to suppress some
tumors in animal studies. Oral administration of L. casei for instance, was effective in suppressing chemically
induced tumors in mice (Matsuzaki, 1998) and the recurrence of superficial bladder cancer in humans (Aso et al,
1995)

Consumption of milk containing cells of L. acidophilus can improve lactose indigestion in humans
classified as lactose maldigestors(Kim & Gilliland, 1983). As part of this study, lactose maldigestors were
subjected to breath hydrogen test (BHT) at seven day intervals using control milk or milk containing cells of L.

acidophilus as the test dose (5 ml/kg body weight) following 12 hour fasts. On day 0 and 7 the test dose was control
milk and on days 14 and 21 the milk containing lactobacilli was the test dose. They were instructed not to consume
milk products between the test days. The results revealed significantly lower levels of breath hydrogen when milk
containing the lactobacilli was the test dose. The benefit was due to the presence of -galactosidase in the L.

acidophilus. To realize the benefit from such a product it is essential that the cells for preparing the product be
grown in a medium containing lactose as the sugar source since -galactosidase is inducible in this organism.
Yogurt containing viable starter cultures has been shown to have similar effects due to -galactosidase content of the
traditional starter cultures (Kim and Gilliland 1984, Kolars et al, 1984). The primary factor required to provide this
benefit is adequate levels of -galactosidase in the bacterial cells.
The above findings show that probiotic bacteria can be used to provide an enzyme(s) that may be useful in
the digestive processes in the intestine. In a related study in our laboratories we have studied the potential of
amylase positive cultures to improve starch digestion following their consumption. Using weaning aged pigs as an
animal model, we tested the influence on weight gain of feeding a culture of L. acidophilus selected for its ability to
hydrolyze starch. The culture was grown in a broth using starch as carbohydrate source. The cells were harvested
and resuspended in nonfat milk for the feeding trial. Two levels of L. acidophilus L23 tested produced significant
increases in daily weight gain during a five-week feeding period compared to the control group. The results likely
were due to the amylase activity of the culture. This presents another potential for a nutritional impact for using a
selected probiotic for young children in developing countries where starchy foods are a staple.
Because of the risk of coronary heart disease and fatal heart attacks occurring in hypercholesterolemic
individuals, there has for a number of years been interest in means whereby serum cholesterol levels could be
reduced in these people. The risk of coronary heart disease can be significantly reduced by lowering serum
cholesterol levels (Lipid Research Clinics Program, 1984).
Serum cholesterol levels can be influenced by the intestinal microflora. For example, germ-free animals on
an elevated cholesterol diet accumulate approximately twice as much cholesterol in blood as do conventional
animals on a similar diet (Eyssen, 1973). The conventional animals excrete more cholesterol in the feces than do the

germ-free animals. This suggests the possibility that microorganisms in the intestinal tract interfere with cholesterol
absorption from the intestines.
In the 1970s, two studies were published which indicated the potential for organisms such as L. acidophilus
to aid in control of serum cholesterol levels in humans. The first of these involved feeding fermented milk to a
group of 25 men (Mann & Spoerry, 1974). The men were fed milk that had been fermented with what was
described as a wild strain of lactobacillus. The fermented milk was consumed for a period of six days (4-5 liters
per day per man). Because of the large intakes of the milk, all men on the trial gained weight and were expected to
exhibit increases in levels of serum cholesterol. While it was not the objective of the study to find a beneficial effect
of the fermented milk, much to the surprise to the investigators, the serum cholesterol levels decreased in all men.
The decrease was greatest in those who gained the most weight suggesting that those consuming the largest portions
of the fermented milk exhibited the greatest decline. Unfortunately, the main organism involved in the fermentation
of this milk was not isolated and identified.
The other study involved the addition of cells of L. acidophilus to infant formula to determine the effect of
the organism on serum cholesterol levels and on the intestinal flora of the infants (Harrison and Peat, 1975). The
serum cholesterol levels in the infants which received formula containing the lactobacilli decreased significantly
during experimental period while those in the control group showed a slight increase.

The decreases were

associated with increased numbers of lactobacilli in the stools of the infants suggesting that the lactobacilli in the
intestinal tract influenced serum cholesterol levels.
Because of these studies, considerable research has focused on the potential of L. acidophilus and related
bacteria to help control serum cholesterol levels. In our laboratories, we have shown that during anaerobic growth

L. acidophilus will remove cholesterol from the laboratory media supplemented with cholesterol and bile salts
(Gilliland et al, 1985). The ability to assimilate cholesterol in this manner varied among strains. In order to test the
theory that L. acidophilus might exert hypocholesterolemic activity in vivo, we used pigs as an animal model. For
this study we felt it necessary to use a strain which originated from the intestinal tract of a pig since there is evidence
in the literature to indicate that the organism exhibits host specificity. Our investigation revealed that a few strains
of L. acidophilus isolated from pigs assimilated little or no cholesterol during growth in the laboratory media while
others assimilated considerable amounts. Because of this wide variation among strains, our feeding trial was
conducted using two strains of the lactobacilli. Lactobacillus acidophilus RP32, which assimilated the greatest

amount of cholesterol, and L. acidophilus P47, which assimilated little or none in the laboratory tests, were selected
for use in the trial. Pigs were fed twice daily a diet high in cholesterol. Additionally each pig in group one (control)
was given 50 ml of milk per day. Pigs in group two were given 50 ml of milk containing 5x1010 L. acidophilus P47
per day and those in group three were given 50 ml of milk containing 5x1010 cells of L. acidophilus RP32 per day.
The milk in these studies was not fermented thus any effect observed would not be due to something produced in the
milk during fermentation prior to consumption.

Analysis of blood samples taken on days 0 and 10 of the

experimental period revealed that the serum cholesterol levels in the control group increased from day 0 to 10. The
group receiving L. acidophilus P47, which did not assimilate cholesterol in the laboratory medium, had levels of
serum cholesterol comparable to those in the control group on both days. However, those in the group receiving L.

acidophilus RP32 had significantly lower serum cholesterol than observed in the control group on day 10. These
data suggest that a strain which assimilates cholesterol during growth in a laboratory medium has the potential of
influencing serum cholesterol level, whereas those which do not assimilate cholesterol during growth in a laboratory
medium do not. Thus, screening the cultures in a laboratory test would be very useful in selecting a strain for use as
a probiotic to aid in control of serum cholesterol.
There have been a number of other research studies that have shown similar potential for L. acidophilus or
related lactobacilli in controlling serum cholesterol levels (Akalin et al, 1997; Danielson et al, 1989; Grunewald,
1982; Jin et al, 1998). Most of these have been done using animal models.
In another of our studies involving pigs (deRodas et al, 1996), with diet induced hypercholesterolemia we
observed that for animals receiving the milk supplemented with lactobacilli the serum cholesterol level declined
sooner and reached the significantly lower level than was observed in the control group. The animals receiving the
milk supplemented with lactobacilli also exhibited lower levels of bile acid in the serum on all days than did those
receiving the milk without lactobacilli, suggesting that the lactobacilli reduced the absorption of bile acid from the
intestinal tract. These findings suggested the possibility that L. acidophilus interfered with the enterohepatic
circulation of bile acids which in itself could be an important factor in reducing serum cholesterol levels.

Lactobacillus acidophilus can deconjugate bile acids. Thus this activity likely resulted in the lower concentrations
of bile acids in the serum of the pigs that received L. acidophilus. Others (DeSmet et al, 1998) have reported the
involvement of bile acid deconjugation in the cholesterol lowering effect of similar probiotic bacteria. Free or
deconjugated bile acids would not be readily absorbed from the small intestine into the blood (Chickai et al, 1987).

Since our main interest in all of this research concerning control of serum cholesterol levels is related to
reducing serum cholesterol levels in hypercholesterolemic humans, we have tested a number of commercially
available strains of L. acidophilus for the ability to assimilate cholesterol during growth in laboratory media
(Gilliland and Walker, 1990). All these strains of L. acidophilus were commercially available in the United States.
A total of seven commercially available cultures were compared to L. acidophilus ATCC 43121 (the one used in our
pig trial). None of them were as active as the latter in assimilating cholesterol from a laboratory medium. Because
of this we undertook a project to isolate new strains of human origin. In this study, isolates were made from sixteen
human volunteers (Buck and Gilliland, 1994). A total of 123 isolates of L. acidophilus were obtained and were
compared for the ability to assimilate cholesterol. Of the isolates compared in this study, eight assimilated more
(although not significantly) cholesterol than did L. acidophilus ATCC 43121 most of the rest assimilated
significantly less. This again shows variation among strains of L. acidophilus with regard to the ability to assimilate
cholesterol during growth in a laboratory medium and thus, variation with regard to the potential for providing a
benefit in controlling serum cholesterol levels. One of these strains, L. acidophilus L 1, has been used in a human
feeding trial involving hypercholesterolemic humans (Anderson & Gilliland, 1999). For this trial, the L. acidophilus
was used along with S. thermophilus to produce a yogurt type product. Consumption of the yogurt caused a
significant (P<.01) serum cholesterol level reduction of 3.2% in the first phase.

A 3% reduction of serum

cholesterol level translates to approximately a twelve-fold reduction in the risk of developing coronary heart disease.
Thus, we were pleased with the results obtained in the feeding trial. The numbers of L. acidophilus, which
developed in the yogurt product during the fermentation period, were not particularly high because this particular
strain does not grow well in milk. We feel that better hypocholesterolemic results could be obtained if we could
develop the product which contains higher numbers of cells of the L. acidophilus.
Conclusions as to the mechanisms whereby L. acidophilus may exert a hypercholesterolemic effect include
both the ability of the organism to assimilate cholesterol during growth and bile salt deconjugation activity. The
latter is most likely due to the potential for increased bile acid excretion from the body due to deconjugation and to
reduced efficiency of absorption of cholesterol from the gastrointestinal tract rather than coprecipitation of
cholesterol with free bile acids as has been suggested by some researchers.

Importance of selection of culture for use as probiotic

To help ensure that the probiotic culture being used has a positive effect, certain requirements are needed.
Generally speaking, these microorganisms should be a normal inhabitant of the intestinal tract. There is evidence in
the literature indicating that many of these bacteria exhibit host specificity. Thus, it is desirable to use a selected
strain of the bacteria that originated in the host species for which the product is to be used. In other words, if it is to
be used as a probiotic for humans, it is highly desirable that the probiotic bacteria originated in human intestines.
One strain should not be expected to produce all potential benefits. Additionally, it is desirable that bacterial species
have a history of use for producing fermented dairy foods. This is important because it makes getting regulatory
agency approval much easier. If they are to grow and function in the intestinal tract, they must have a certain degree
of bile tolerance. The bile resistance, based on relative ability to grow in the presence of bile acids, varies greatly
among strains of each species of probiotic bacteria. At present time we do not know the minimum level of bile
tolerance required. However, I suggest use of the most bile resistant strain that will provide the desired beneficial
effect. The culture being used must be stable during production and storage of the food containing the probiotic
organism. The most important characteristic of course is that the culture will produce the desired effect. Thus,
some sort of laboratory screening test for the desired effect would be helpful. We have shown, for instance, in the
case of using a probiotic bacteria for control of serum cholesterol levels, that a strain which assimilates cholesterol
during growth in the laboratory medium had much better effect compared to one that assimilated little or no
cholesterol from the broth media.

Stability of probiotic bacteria in milk

In situations where the probiotic bacteria in the form of a concentrated culture are added to milk, we have
shown that the pH at which the cells of L. acidophilus had been grown influenced their stability and survival in the
milk during refrigerated storage (Gilliland and Rich, 1990). In these experiments the culture was grown in a broth
medium, harvested by centrifugation and concentrated in the small volume of milk then frozen in liquid nitrogen.
Such a production and freezing process had no detrimental effect on the cultures. However, growth of two strains of

L. acidophilus at pH 5 resulted in cultures which were more stable during refrigerated storage in milk than were the
same cultures which had been grown at higher pH levels. Thus, the method of growing the culture of probiotic
bacteria, which are added to milk as a dietary adjunct, can have a major impact on survival of the culture during
storage.

10

In the case of B. longum, growth of the culture in broth also had no detrimental effect on the culture during
frozen storage (Reilly and Gilliland, 1999). However when added to milk, of the four strains tested, one (S9) which
had been grown at either pH 5.5, 6.0, 6.5 or 7.0 did not exhibit any decrease in viability during subsequent storage of
the milk at 5C for 21 days. The other three strains were less stable during refrigerated storage. These data indicate
variability among the strains with respect to storage stability.
When cells of L. acidophilus from a frozen concentrated culture were added to cultured yogurt, to provide
an initial population in the range of 1 to 7x107 per gram, there was significant variation among strains with respect
to the maintenance of viability during 28 days of storage at 7C (Nighswonger et al, 1996). Out of four strains
tested, strain L1 exhibited best survival. The experiment was replicated in yogurt made using two different yogurt
starter cultures. Lactobacillus acidophilus L1 exhibited no loss in survival during storage in yogurt made with one
of the starter cultures for 28 days while it did exhibit a significant decline after 28 days in the other. Not only was
there variation among the strains of L. acidophilus with regard to stability in yogurt, but there appeared to be
difference between the two yogurt cultures with respect to their influence on stability of L. acidophilus during
refrigerated storage. Thus, selection of the yogurt culture may also be important in helping assure that adequate
viable numbers of the probiotic bacteria are maintained during storage of the product.
Preparation of dried milk products containing probiotic bacteria offers a great challenge. Spray drying of
milk containing added cells of L. acidophilus can be very detrimental to viability of the organism. Espina and
Packard (1979) compared the influence of spray drying at three outlet temperatures (75 C, 80 C and 85 C) milk
adjusted to 25% and 40% nonfat solids containing added cells of L. acidophilus. Best survival was obtained in the
25% solids milk using an outlet temperature of 75 C. Under these conditions, greater than one log cycle of viability
was lost yielding a population of 9.8 x 107 /g in the final product. No storage stability tests were done. King and
Lin (1995) reported 91-96% survival during freeze drying of cells of L. acidophilus suspended in 10% reconstituted
nonfat milk supplemented with 5.34 mg Ca++ and 41.5 mg glycerol per ml. No storage stability data was reported.
Only one strain of L. acidophilus was included in each of these studies. Based on variability among strains for other
characteristics we should expect variation among strains with respect to survival during spray drying and subsequent
storage of dried products containing probiotic bacteria. The composition of the medium and/or conditions in which
the cultures are grown prior to drying likely would influence their survival.

11

Even though not published, the industry that produces starter cultures possess the knowledge and
technology to stabilize cultures to maximize survival during drying and storage. The best approach to producing a
dried milk containing probiotic bacteria probably will involve blending the freeze dried probiotic culture with milk
powder.

Recommendations
If powered milk products containing probiotic bacteria are to be supplied for developing countries the
potential benefits of controlling intestinal infections and improving nutrient (primarily lactose) utilization should be
considered. To achieve this may require providing more than one strain and/or species of probiotic bacteria in the
product. One strain of one species should not necessarily be expected to provide more than on benefit. The
culture(s) should be selected on the basis of being able to provide the desired result. It also must be stable during
drying and storage in the milk powder. This might be accomplished using existing cultures or may involve the need
to isolate and develop new strains. The biggest challenge will be ensure stability during storage of the dried product.

Research needs
1.

Selection of probiotic to produce desired effect. This should involve laboratory screening test(s)
as well as clinical trials.

2.

Evaluation of stability of the selected probiotic organisms during drying and storage in powdered
milk.

3.

Determine best way to deliver the powdered milk containing probiotic. This likely will involve
the importance of packaging material as well as size of package.

12

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