Effect of Fermentation on B-Vitamin Content
of Milk in Sweden
ABSTRACT
Samples of milk and fermented milk
products were analyzed for B vitamin
content by standard chemical and micro-
biological assay procedures. Fermented
milk products showed an increase in folic
acid content (ropy milk exhibited a
twofold increase) and a slight decrease in
concentration of vitamin Blz. Orotic acid
was reduced in all fermented milk prod-
ucts (buttermilk 46.0%, yogurt 47.8%,
kefir 17.7%, ropy milk 36.5%, and
acidophilus milk 22.5%). Other vitamins
were affected only slightly.
INTRODUCTION
Cow's milk is a good source of most vitamins
(21) except ascorbic acid and vitamin B12. The
nutritional value of fermented milk products is
assumed to be similar to the milk from which
they are made (45, 49). However, fermentation
may lead to an increased concentration of
B-vitamins, but results reported are contra-
dictory (11, 15, 28, 30, 50).
With increasing demand for more nutritious
foods, knowledge of changes in B-vitamin
content caused by fermentation is important.
Lactic acid bacteria in fermented milk products
has been suggested as a possible supplementary
source of vitamins in human nutrition (31, 51).
During fermentation there is a dramatic
increase in cell population representing both
colony forming units as well as dormant ceils.
The increase continues, though slower, even
during storage.
The B-vitamins are components of coenzymes
and are extremely important in the metabolism
and energy utilization of the cell and more or
less essential for starter cultures used in fermen-
tation of milk (13). Starters show various needs
and ability to synthesize B-vitamins in excess.
Received December 4, 1980.
1982 J Dairy Sci 65:353-359
LIVIA ALM
Aria
S-105 46 Stockholm, Sweden
Microbial biosynthesis of B-vitamins is a useful
method for improving the B-vitamin content of
dairy products (21). To assess the degree of this
ability was one of the targets for study.
Vitamin Bl3, or orotic acid (OA), (6) is a
growth factor for lactobacilli (2, 39, 40) and
other microorganisms (12) and is essential for
normal development of the intestinal flora
(24, 25). In cow's milk, OA is relatively high
(31, 32). It is believed to promote development
of the rumen flora of the calf. The OA content
in cow's milk is about 70 rag/liter (14, 18, 37,
38) whereas the content of OA in human milk
is low (25, 46).
In view of conflicting reports in the litera-
ture on the influence of fermentation on
B-vitamins we were interested to evaluate the
content of the B-vitamins in Swedish fermented
milk products.
MATERIALS
AND METHODS
Products investigated were nontreated pool-
ed farm milk, regular milk with 3.0% (wt/vol)
fat, buttermilk (filmj~51k) with 3.0% (wt/vol)
fat, yogurt with 3.0% (wt/vol) fat, kefir with
3.0% (wt/vol) fat, ropy milk with 3.0% (wt/vol)
fat, sweet buttermilk with .6% (wt/vol) fat,
sour (fermented) buttermilk with .6% (wt/vol)
fat, ultra-high heat treated (UHT) low fat milk
with .5% (wt/vol) fat, low fat acidophilus milk,
special V-medium, acidophilus milk, and bifidus
milk manufactured from V-medium.
Production Methods
Nontreated pooled farm milk has no stand-
ardized fat content, is not homogenized, and is
not heat treated.
Regular milk. Cow's milk is standardized to
3.0% (wt/vol) fat and pasteurized (75°C for 15
s), pH range 6.7 to 6.8.
Buttermilk. Cow's milk, 3.0% (wt/vol) fat, is
heat treated (92°C for 3 min), cooled, inocu-
lated with a commercial mixture of Streptococ-
cus lactis, Streptococcus cremoris, Streptococ-
cus diacetylactis, and Leuconostoc cremoris
353
354 ALM

TABLE 1. Microbial strains in vitamin assay.

Vitamin assayed Test organism

B2, Riboflavin Lactobacillus casei (ATCC 7469)

B6 , Pyridoxine Saccbaromyces carlsbergensis (ATCC 9080)
Pantothenic acid Lactobacillusplantarurn (ATCC 8014)
Biotin Lactobacillus plantarum (ATCC 8014)
B12, Cobalamine Lactobacillus leicbmanii (ATCC 7880)
Folic acid Lactobacillus easel (ATCC 7469)
Tryptophant Lactobacillus plantarum (ATCC 8014)
Nicotinic acid Lactobacillus plantarum (ATCC 8014)

1Tryptophan may be converted into niacin equivalents, in the ratio 1:60; 60 mg tryptophan = 1 mg niacin.

(citrovorum), and cultured at 20°C for 20 to 24 and cultured at 37°C for 40 h, pH range 4.3 to
h, pH range 4.4 to 4. 5. 4.5.
Sweet buttermilk. Nonfermented cream, Bifidus milk. V-medium is inoculated with
40% (wt/vol) fat, is churned to obtain butter. Bifidobacterium bifidum (Lactobacillus bifidus
After churning, the sweet buttermilk has a fat (NCDO 11863) and cultured at 37°C for 40 to
content of .7% (wt/vol) and pH range 6.7 to 50 h, pH range 4.2 to 4.4.
6.8.
Fermented (sour)buttermilk. Nonfermented Handling of the Samples
cream, 40% (wt/vol) fat, is inoculated with a Samples either were taken directly from
commercial mixed culture of Streptococcus commercial production and analyzed immedi-
lactis, Streptococcus cremoris, Streptococcus ately or preserved before analysis by lyophiliza-
diacetylactis, and Leuconostoc cremoris (citro- tion. Before analysis, the lyophilized samples
vorum), cultured at 20 to 21°C for 20 to 24 h, were reconstituted to the original state by the
and churned to obtain butter. The fermented addition of 9 volumes distilled water. No
buttermilk has a fat content of .7% (wt/vol) differences in vitamin contents between non-
and a pH range of 4.4 to 4.5. preserved and lyophilized and reconstituted
Kefir. Cow's milk, 3.0% (wt/vol) fat, is heat
treated (92°C for 3 min), cooled, inoculated
with a commercial mixture of kefir grains, and
cultured at 20°C for 18 to 20 h, pH range 4.3 TABLE 2. B-vitamins in noninoculated milk. (Each
to 4.5. mean was from observations on four samples from
Ropy milk. Cow's milk, 3.0% (wt/vol) fat, is different batches.)
heated (90°C for 3 rain), cooled, inoculated
with a commercial mixture of Streptococcus Vitamin X SD Units
lactis var. longi and Leuconostoc cremoris
(citrovorum), and cultured at 17 to 18°C for 20 B1, Thiamine .34 .03 mg/liter
to 22 h, pH range 4.5 to 4.6. B:, Riboflavin 1.16 .01 mg/liter
B6 , Pyridoxine .38 .01 mg/liter
V-medium. Lowfat cow's milk, .5% (wt/vol) Pantothenic acid 2.6 .20 rag/liter
fat, added with .5% (wt/vol) soybean oil, pH Biotin 14.0 1.0 tag/liter
6.7 to 6.8, is heat treated (75°C for 15 s) and Folic acid 37.0 8.0 /~g/liter
fortified with a protein concentrate from skim BI2., Cobalamine 3.1 .3 #g/liter
milk. In this way the protein of the milk Nicotinic acid .68 .01 mg/liter
Tryptophant 308.0 10.0 mg/liter
formula was increased from about 3.4 to 5.0%. Niacin equivalents 5.8 .2 mg/liter
The mixture was allowed to swell for 15 to 18 B~.~,Orotic acid 67.1 .7 rag/liter
h at 4°C and then was heated at 140°C for 3 to
4s. 1Tryptophan may be converted into niacin equi-
Acidopbilus milk. V-medium is inoculated valents, in the ratio 1:60; 60 mg tryptophan = 1 mg
with Lactobacillus acidopbilus (NCDO 1748) niacin.

Journal of Dairy Science Vol. 65, No. 3, 1982

 TABLE 3. B-vitamins in milk and fermented milk products. Data compiled from literature.

Thiamine Riboflavin Pyridoxine Pantothenic Cobalamin Nicotinic

Sample B1 B2 B6 acid Biotin Folic acid B12 acid References

(mg/liter) 0~g/liter) (mg/liter) '~

Milk .43 1.7 .48 3.6 40 55 4.5 .95 47 ~'~
.43 1.7 .60 3.4 30 59 4.2 .93 52
Buttermilk .34 1.6 .40 2,0 1.0 47
.42 1.7 .35 3.8 11 110 2.3 0.55 52
Milk + growth factor 4.8 41 40 4.0 1.30 45
Milk w i t h o u t 4.02 36 20 4.0 1.09 45
and added with .40 1.77 .38 68 2.0 1.03 19, 30
growth factor 3.0 26
Yogurt .37 2.0 .50 100 1,0 1.5 47, 22 Z
.37 1.4 .35 12 1.2 1.3 52
Yogurt 1.5 3.8 39 40 3.0 1.3 45, 3
cultured .5 1.1-2.6 87-138 2.0 23, 26, 27
Yogurt
acidified 4.3 43 40 4.0 1.3 45
Kefir milk .43 1.40 .27 2.5 22 60 1.1 1.08 29
.17 22 36
Kefir day 1 .56 1.17 .31 2.45 15 10.5 .8 1.08 29
~q
Kefir day 4 .47 1.25 .34 2.57 17 92 .9 1.18 29
Kefir day 7 .50 1.39 .39 2.56 17 82 1.4 1.11 29

O
Ox

Z
o

0o
bo
356 ALM

samples were significant. Analyses were on

samples pooled from 10 batches of fermented
products.

Methods of V i t a m i n Assay
Vitamin profile, %
+140.l l
Thiamine (B1) was measured by a fluoromet- +100: i
ric method (4). Riboflavin (B2) was measured +80. i
by two methods, a fluorometric method (4, 33) +60 Buttermilk
and a microbiological assay (43). Orotic acid +40.
(OA) was measured by a spectrophotometric +20.
method according to Adachi (1). Other B- +0
vitamins were measured by microbiological -20 1 111Days
assay with microorganism strains in Table
+1207
1. The procedure for analysis is described ~100~
in detail in Pharmacopoea Nordica (43).
+80 i
+60 Yoghurt
I
RESULTS +4oi
i

i i
+20i
V i t a m i n Content of Milk -+0-~
The B-vitamin in noninoculated milk is in -207
-40 - ill
Table 2. As compared to averages reported
I
1 -il
I
11!1
,
1111111111111,1 11 Days
(Table 3), vitamin contents were low.
+40] ! Ke~ir i I i
+201 i i
Changes in V i t a m i n Content Following Fermentation +0" i_ ! ! I I a-
Figures 1 and 2 show B-vitamin concentra- _2oJ 1 "
+223- 111 iii
I! '
Ii
-i1
11 'l '11" ll Days
11 1 11 1
tions of fermented products at different stages
of storage. Concentrations are percent changes +160 -~
i
of B-vitamin of nonfermented milk. Vitamins +120-"

lil
B6, B12, and biotin decreased in buttermilk by +80-"
about 15% whereas folic acid increased by +60- i
Ropy milk
about 140%. Pantothenic acid, biotin, and B12 +40-
decreased in yogurt; folio acid increased also in +20 -
this product by about 120%. Vitamin B6, B12,
and biotin decreased by 15% in kefir. Folic acid -20 -
increased by nearly 40% day 1, although its -40 -
-60- 1 I 1 13 13 1 13,1 13, 1 13!Days
content was reduced to 25% of nonfermented
milk after storage for 11 days.
+100]
Minor changes of vitamin BI, B~, B6, and
biotin in ropy milk were seen. Folic acid
+801
+601 Sour buttermilk I
increased more than two times after one day's
+401
fermentation. During storage for 13 days, its

content decreased to about 120% of nonfer-
mented milk. Vitamin Bl: decreased continu-
ously (by about 20% and 40% on days 1 and
13).
Vitamin B1 and folic acid content increased
by about 20% and 100% in sour buttermilk.
Concentration of other B-vitamins showed only
minor alterations.
The only milk product where folic acid did
n o t increase during fermentation was acidophi-
lus milk (Figure 2). When acidophilus milk was
-;°Jl@
+2Oll
+O/m
i 1 '1
® ,@ i® @
Figure 1. The B-vitamin profile in buttermilk,
yogurt, kefir, ropy milk, and fermented sour butter-
milk. Percent increase or decrease compared with the
milk from which the products were made. 1 = B,,
thiamine; 2 = B2 , riboflavin; 3 = B6, pyridoxine;
4 = pantothenic acid; 5 = biotin; 6 = folic acid; 7 =
B,~, cobalamin; 8 = niacin equivalents.

Journal of Dairy Science Vol. 65, No. 3, 1982

 B-VITAMINSIN FERMENTED MILK 357

manufactured from low fat milk, the concen- yogurt, the concentration of thiamine decreased
tration of vitamin B1 increased by about 30% whereas riboflavin remained unchanged. Blanc
whereas the content of the other B-vitamins (7, 8) and others (3) reported increased thiamine
only showed minor changes. However, when and riboflavin, whereas Koser (34) reported
acidophilus milk was made from V-medium, an decreases of both these B-vitamins in yogurt.
entirely different vitamin profile was observed. The pyridoxine content in yogurt remained
The concentration of folic acid decreased by 20 almost unchanged during fermentation whereas
to 40% and that of vitamin B12 by 40 to 60%. Blanc (8) reported a decrease. Nicotinic acid
The B-vitamin profile of bifidus milk also is increased during fermentation (10, 36), but our
shown in Figure 2. The most pronounced investigation failed to confirm this result. The
changes were an increase in folic acid concen- varying results reported by different investiga-
tration by about 100% and a decrease in the tors may reflect different metabolic activities of
content of B12 (to about half the original fermenting microorganisms.
mean) after 13 days of storage. The general increase in folic acid following
The OA in milk and fermented milk prod- fermentation agrees well with findings reported
ucts are in Table 4. Concentration of OA by other authors (17, 27, 45). Also, the general-
decreased significantly in all samples. The most ly lower OA in fermented milk products than in
pronounced reduction was for" yogurt and the original milk seems to be a common finding.
buttermilk where OA content decreased by The particularly high consumption of OA by
more than 45%. yogurt starters (cf. also 18, 41, 48) might be
related to the relatively high fermentation
DISCUSSION temperature (42 to 45°C) and the relatively
Fermentation affects most B-vitamins. In high cell number per milliliter in yogurt pro-
duction.
Milk and milk products are the only sources
of OA in the human diet. The OA affects lipid
Vitamin profile, %
metabolism in experimental animals (9, 49) and
+4o~ | Acidophilusmilk[.
[
I i !
humans (5, 49). In large amounts, OA may
+2otl promote the development of fatty liver (5, 35,
-+0{" 42). Hence, it may be questioned whether OA
-20Jl 1 il I~ Day
is of any nutritional value or whether it repre-
Acidophilusmilk sents a dietary component that can only be
+20 - I
I
+-0- tolerated in limited amounts, especially in
-20 -
F infants. If this is the case, the reduction in OA
-40
-60- 1 12: Days
+100-
+80-
+60-
} TABLE 4. Orotic acid content in milk and fermented
Bifidusmiik products.
+40-
+20- Samples Orotic acid
'+0- n=5 (rag/liter)' % Reduction
-20-
-40-
- I ! 1 13jDaySlvit. SD
-60- L 13-1 1311(~)1311 13'1
[ 13i1! [
Regular milk 67.1 .41
® Buttermilk 35.9 .83 46~5
Yogurt 35.0 .88 47.8
Figure 2. The~B:vitamin profile in lowfat acid0phi- Kefir 55.2 .96 17.7
lus milk, acidophilus milk, and bifidus milk made from Ropy milk 42.6 .66 36.5
V-medium. Percent increase or decrease compared Acidophilus milk 52.0 4.17 22.5
with the milk from which the products were made.
I = B , , thiamine; 2=B2, riboflavin; 3 = B e, pyri-
doxine; 4 = panthothenic acid; 5 = biotin; 6 = folic i AII means for fermented milk products differ
acid; 7 = B,2, cohalamin; 8 = niacin equivalents. (P<.001) from regular milk.

Journal of Dairy Science Vol. 65, No. 3, 1982

358 ALM
c o n t e n t during f e r m e n t a t i o n m a y be considered
an advantage f r o m a nutritional standpoint.
An i m p o r t a n t question concerning B-
vitamins in f e r m e n t e d milk p r o d u c t s is their
bioavailability (39, 44). The vitamins f u n c t i o n
as integrated parts of proteins (13, 20) and,
therefore, m a y occur b o t h in free f o r m and as
cell-bound constituents in f e r m e n t e d milk. It is
n o t k n o w n w h e t h e r or where lysis o f lacto-
bacilli occurs during the digestion process
and w h e t h e r cell-bound vitamins are m a d e
available to the h o s t (24). Possibly s o m e t y p e
o f e n z y m e s derived f r o m the normal intestinal
flora participate in the release o f cell-bound
vitamins, and it m a y be speculated t h a t one
way in which lactobacilli m a y influence the
metabolic activities of the normal flora (16, 19)
is by supplying it with B-vitamins. The bioavaJl-
ability of dietary B-vitamins is a subject w o r t h y
of further investigation.
In summary, f e r m e n t a t i o n o f c o w ' s milk
usually results in relatively m i n o r changes in the
c o n c e n t r a t i o n of B-vitamins, e x c e p t in the cases
of folic acid and orotic acid where the concen-
trations are significandy increased (folic acid)
or decreased (orotic acid). F u r t h e r studies are
needed to evaluate nutritional implications of
these changes.
ACKNOWLEDGMENTS
This w o r k was s u p p o r t e d by grants f r o m the
Swedish Dairy Association, SMR, Sweden. The
a u t h o r is i n d e b t e d to J a n - A k e Gustafsson for
his e n c o u r a g e m e n t and help in preparing this
manuscript.
REFERENCES
1 Adachi, T., A. Tanimura, and M. Asahina. 1963. A
colorimetric determination for orotic acid. J.
Vitaminol. 9:217.
2 Ahmed, A. A., G. A. Porter, and R. D. McCarthy.
1978. Isolation, quantification and biosynthetic
origin of orotic acid in milk. J. Dairy Sci. 61 : 39.
3 Anagama, Y. 1971. Vitamin B12 in yoghurt from
markets in Japan. Res. Bull. Fac. Gifu Univ.
31:291.
4 Association of Official Analytical Chemists. 1975.
Official methods of analysis, l lth ed. AOAC,
Washington, DC.
5 Bernstein, B. A., T. Richardson, and C. H. Amund-
son. 1975. Inhibition of cholesterol biosynthesis
by bovine milk, cultured buttermilk and orotic
acid. J. Dairy Sci. 59:539.
6 Biscarro, G., and E. Bellone. 1904. Ueher die
Orots~iure. (Orotic acid.) Ann. Soc. Chim., Milano
11:18.
Journal of Dairy Science Vol. 65, No. 3, 1982
7 Blanc, B. 1966. Influence du stockage et des
traitements industriels sur la teneur en vitamines
du lait de vache. (The influence of storage and
industrial treatment on the content of vitamins in
cow's milk.) Le Lait 46:141.
8 Blanc, B. 1973. Der Wert der Sauermilchprodukte
in der modernen Emghrung. (The importance of
fermented milk products in modern nutrition.)
Schweiz. Milchztg. 99:463.
9 Boguslawski, W., and J. Wrobel. 1974. An inhibitor
of sterol biosynthesis present in cow's milk. Nature
247:210.
10 Brochu, E., R. Riel, and C. Vezina. 1959. Les
bacteries lactiques et les laits fermentts. (Lactic
acid bacteria and fermented milk.) Rech. Agron.
No 3, Le Conseil des Recherches Agricoles, Minis-
t~re de L'Agriculture, Quebec.
11 Cerna, J., and H. Hrabova. 1977. Biologic enrich-
ment of fermented milk beverages with vitamin
B12 and folic acid. Milchwissenschaft 32:247.
12 Chert, M-H., and B. L. Larson. 1971. Pyrimidine
synthesis pathway enzymes and orotic acid in
bovine mammary tissue. J. Dairy Sci. 54:842.
13 Cormier, M. 1977. Regulatory mechanisms of
energy needs: vitamins in energy utilization.
(Review). Prog. Food Nutr. Sci. 2:347.
14 Deutsch, A., and S. Mattson. 1960. Page 1 in
Acid-soluble nucleotides in cow's milk and colos-
trum. Rep. No. 63 Milk Dairy Res., Alnarp, Swe-
den.
15 Dolezalek, J., and V. Havlikova. 1972. Possibilities
of utilization of propionibacteria in the production
of Kefir. Vysoke skoly chemikotechnologicke v
Praze, E 33 Potraviny. Sci. Papers, Inst. Chem.
Technol., Prague E 33.
16 Drasar, B. S., and L. J. Hill. 1974. Human intes-
tinal flora. Academic Press, New York, NY.
17 Emanuilov, I., and L. Nachev. 1957. Vitamin B12
content of Bulgarian sour milk. (Izv. Mikrobiol.
Inst., Sofiya, 8:283.) Dairy Sci. Abstr. 20(1958).
Abstr. No 1936.
18 Gajos, E., and H. Kriezliewicz. 1974. Estimation of
orotic acid in milk and milk products. Przegl.
Mlecz. 23:323 (Suppl. 1):7.
19 Glass, L , and T. I. Hedrick. 1976. Bacterial growth
and vitamin content of milk. J. Milk Food Technol.
39:325.
20 Goldin, B. R., L. Swenson, J. Dwyer, M. Sexton,
and S. L. Gorbach. 1980. Effect of diet and
Lactobacillus Acidopbilus supplements on human
fecal bacterial enzymes. J. Nat. Cancer Inst.
64:255.
21 Gregory, M. E. 1975. Water-soluble vitamins in
milk and milk products. J. Dairy Res. 42:197.
22 Gulmezoglu, E., and N. H. Fisek. 1956. Riboflavin,
nicotinic acid and biotin content of yoghurt.
Turkish Bull. Hyg. Biol. 16:222.
23 Gorner, F., and V. Oravcova. 1969. Changes in
thiamine and riboflavin contents during yoghurt
and Gouda cheese fermentation. (Pol'nohospo-
darstvo 15:825.) Dairy Sci. Abstr. 33 (1971).
Abstr. No 1607.
24 Haenel, H. 1958. Darmflora und Vitaminhaushalt.
(Intestinal flora and vitamin balance.) Erniflarungs-
forschung 3:282.
 B-VITAMINS IN FERMENTED MILK
25 Hallanger, L. E. 1953. Orotic acid in milk. J. Biol.
Chem. 99-103:83.
26 Hegedfis, M. 1972. Elelmizserek B12 vitamintar-
talmfinak meghat~roz~sa mikrobiologiai modszerrel.
II. Elelmiszerek B~2 vitamintartalmanak vizsg~lata.
(Determination of BI~ using microbiological
methods. II. The content of B~2 in various foods.
Elelmiszerv. K6zlem~nyek, 18:19.
27 Hoppner, K., B. Lampi, and D. E. Perrin. 1972.
The free and total folate activity in foods available
on the Canadian market. Can. J. Food Technol.
5:60.
28 Joe, A. M., K. M. Shahani, and A. Kilara. 1975.
Vitamin B content of cultured and acidified sour
cream. J. Dairy Sci. 58:791.
29 Karlin, R. 1958. Sur l'enrichissement du kefir en
vitamine BI: par l'adjonction de Propioniibac-
terium sbermanii. (Increased levels of B12 in kefir
by addition ofPropioniibacterium shermanni.) Int.
Z. Vitaminforsch. 12:358.
30 Karlin, R. 1969. On the content of folates in bulk
milk. Effects of various heat treatments on levels
of folates. Int. Z. Vitaminforsch. 39:359.
31 Kiermeier, F., and A. Buckl. 1968. Einfiiisse auf
der Orotsiiure Gehait in Kuhmilch. (Influence on
orotic acid content in milk.) Z. Lebensm. Unters.
forsch. 138:284.
32 Kobata, A., S. Ziro, and M. Kida. 1962. The acid
soluble nucleotides in milk. J. Biochem. 51:277.
33 Kon, S. K., and S. Y. Thomson. 1957. Measure-
ments of vitamins in the control of milk processing.
Milchwissenschaft 12:166.
34 Koser, S. A. 1968. Vitamin requirements of
bacteria and yeasts. Charles Thomas Publ., Spring-
field IL.
35 Larson, B. L., and H. M. Hegarthy. 1977. Orotic
acid and pyrimidine nucleotides of ruminant
milks. J. Dairy Sci. 60:1223.
36 Lasitsa, O. J. 1967. Pyridoxine content of different
types of milk and kefir. (Paediatria Akush. Ginekol.
1:23.) Dairy Sci. Abstr. 29(1967). Abstr. No
2047.
37 Levenson, S. M., and B. Tennant. 1963. Contribu-
tion of intestinal flora to the nutrition of the host
animal. Some metabolic and nutritional studies
with germfree animals. Fed. Proc. 22:109.
38 Munchberg, F., G. Tsompanidou, and R. Leskova.
1971. Untersuchung ilber das Vorkommen der
Orots'~iure in der Milch. (Determination of the
359
orotic acid content in milk.) Milchwissenschaft
26:210.
39 Nelson, E. W., R. R. Streiff, and J. J. Cerda. 1975.
Comparative availability of folate and vitamin C
from a synthetic and natural source. Am. J. Clin
Nutr. 28:1014.
40 O'Donovan, G. A. and J. Neuhard. 1970. Pyrimidine
metabolism in microorganisms. Bacteriol. Rev.
34:275.
41 Okonkvo, P. O., and J. E. Kinsella. 1969. Orotic
acid in yoghurt. J. Dairy Sci. 52:1861.
42 Okonkvo, P. O., and J. E. Kinsella. 1974. Fatty
livers induction by orotic acid content in skim
milk powder. Experientia 30:993.
43 Pharmacopoea Nordica, 1963. Microbiological
assay of vitamins. Pharmacopoea Nordica, Vol IV
(1963).
44 Polansky, M. M., and E. M. Toepfer. 1969. Vitamin
B6 components in meats, fish, dairy products and
commercial infant formulas. J. Agric. Fed. Chem.
17:1394.
45 Reddy, K. P., K. M. Shahani, and S. M. Kulkarni.
1975. B-complex vitamins in cultured and acidified
yoghurt. J. Dairy Sci. 59:19.
46 Reif, G. D., K. M. Shahani, J. R. Vakil, and L. K.
Crowe. 1976. Factors affecting B-complex vitamin
content of cottage cheese. J. Dairy Sci. 59:410.
47 Renner, E. 1974. Milch und Milchprodukte in der
Ernghrung des Menschen. (Milk and milk products
in human nutrition.) Arbeitsgemeinschaft fiir das
Milchwirtschaftliche Fachbuch. Hitdesheim. Volks-
wirtsch. Verlag.
48 Ritter, W. 1977. Die quantitative Bestimmung der
Orotsiiure als M6glichkeit zur Ermittlung des
Milchanteils in Lebelsmitteln. (The quantitative
determination of orotic acid and its possible use as
an indicator of the level of milk additives in food.)
Mitt. Geb. Lebensmittelunters. Hyg. 68:240.
49 Richardson, T. 1978. The hypercholesteremic
effect of milk. J. Food Prot. 41:226.
50 Shahani, K. M., and C. R. Chandan. 1979. Nutri-
tional and healthful aspects of cultured and culture-
containing dairy foods. J. Dairy Sci. 62:1685.
51 Shahani, K. M., and A. D. Ayebo. 1980. Role of
dietary lactobacilli in gastrointestinal microecology.
Am. J. Clin. Nutr. 33:2448.
52 Webb, B. H., A. H. Johnson, and J. A. Alford.
1975. Fundamentals of dairy chemistry. 3rd ed.
Avi Publ. Co., Westport, CT.
Journal of Dairy Science Vol. 65, No. 3, 1982