BREEDER MALE:FEMALE RATIOS 1575

Snedecor, G. W., 1956. Statistical Methods. 5th Ed., breeders. Br. Poultry Sci. 12: 327-331.
Iowa State College Press, Ames, Iowa. Woodard, A. E., and H. Abplanalp, 1967. The effects
Wilson, H. R., and R. H. Harms, 1971. Male to female of mating ratio and age on fertility and hatchability
ratios for broiler-type and egg production-type in Japanese quail. Poultry Sci. 46: 383-388.

Fermentation of Whole Egg by

Heterofermentative Streptococci12
S. J. GALLUZZO, O. J. COTTERILL AND R. T. MARSHALL
Department of Food Science and Nutrition, University of Missouri-Columbia, Columbia, Missouri
65201
(Received for publication December 14, 1973)

ABSTRACT Growth and diacetyl production of heterofermentative streptococci in liquid

whole egg (WE) were investigated. Three mixed-strain lactic cultures, a Streptococcus diacetilactis
culture, and two Leuconostoc species did not grow well in pasteurized (60° C. for 3.5 min.)
WE with or without added yeast extract. Lowering the pH and/or additional heating of the
WE enhanced growth. S. diacetilactis grew in pasteurized WE when pH was reduced to 6.5
or below. Both superheating (65° C. for 20 min.) the WE and pH adjustment were necessary
for good growth of the other cultures.
S. diacetilactis produced most diacetyl in WE acidified to pH 5.5 with citric acid. Peak
concentrations of 9 p.p.m. were noted after 8 hr. of fermentation at 21.5° C. A "buttery"
aroma was readily apparent. Diacetyl concentrations were low and varied in WE acidified
with citric acid, when inoculated with mixed-strain cultures. Leuconostoc cultures produced
a mild fermented odor without diacetyl. When lactic acid was used for pH adjustment, the
mixed strain cultures and S. diacetilactis produced a "metallic" off-odor which increased with
fermentation time. It was possible to simultaneously desugar WE and impart flavors and /or
aromas characteristic of dairy products by fermentation with S. diacetilactis. However, growth
of a bacillus contaminate during fermentation was a problem.
POULTRY SCIENCE 53: 1575-1584, 1974

INTRODUCTION process utilizing S. lactis, S. diacetilactis,

F ERMENTATION of egg products for the
development of new products has not
been reported. The heterofermentative
streptococci show promise. Previously, this
group of organisms has received attention
for desugaring egg products prior to drying.
Stewart et al. (1943) desugared egg white
by inoculating the liquid with a pure culture
of streptococci. Some flavor changes were
indicated. Hopkins et al. (1947) patented a
1. Contribution from the Missouri Agricultural Ex-
periment Station. Journal Series Number 6867.
2. Presented at the 62nd Annual Meeting of the
Poultry Science Association, Inc. South Dakota State
University, Brookings (see Poultry Science 52: 2030,
1973).
and S. thermophilus to desugar egg products.
They claimed that these organisms rapidly
desugared whole egg but failed to remove
a like amount of sugar from egg white. Kaplan
et al. (1950) studied desugarization of egg
white by fermentation with resting cells of
S. lactis. Ayres (1958) found that S. lactis
decreased in numbers in egg white unless
large inocula were used and yeast extract
added. Bean et al. (1963) reported satisfactory
desugaring rates of liquid egg white by strains
of S. cremoris and L. mesenteroides. Fer-
mentations at low pH (5.0) were apparently
feasible with L. casei, S. lactis, and M.
freudenreichii.
This study concerns growth and diacetyl
production by heterofermentative strepto-
1576 S. J. GALLUZZO, O. J. COTTERILL AND R. T. MARSHALL
Magnetic stirring
bar
Magnetic stirrer
FIG. 1. Apparatus for heating normally pasteur-
ized and superheated whole egg.
cocci in whole egg. These organisms are
important acid and flavor producers in the
lactic fermentation of buttermilk, sour cream,
cottage cheese, and cultured butter.
MATERIALS AND METHODS
Liquid Whole Egg. Whole egg (WE) was
prepared aseptically as described by Cotterill
(1968), except that an acid iodophor rather
than hypochlorite was used to sanitize the
shells.
Heat Treatments. Blended and filtered WE
(aseptically prepared) was heat treated in a
sterile Erlenmeyer flask fitted with a rubber
stopper and a thermometer (Fig. 1). Initial
water bath temperature was set 2-3° C. higher
than the final product temperature so that
final water and WE temperatures equilibrated
at the desired holding temperature. The WE
was stirred vigorously during heating with
a magnetic stirring bar. Two heat treatments
(holding conditions) were used: (1) normal
pasteurization at 60° C. for 3.5 min.; and (2)
"superheating" at 65° C. for 5, 10, 15, and
20 min. No visible coagulation was apparent.
Come-up times ranged from about 7 min. in
125 ml. flasks to about 10 min. in 500 ml.
flasks. Following heat treatment, the ther-
mometer and stopper were removed, and a
sterile piece of foil was quickly placed over
each flask. Flasks were then placed in an
ice water bath and cooled prior to plating
of contents. Heated WE was held overnight
at 2° C , and only samples with 5 cells/ml.
or fewer were used for inoculation with
experimental cultures.
Cultures and Media. Six cultures were uti-
lized throughout the study. A Streptococcus
diacetilactis culture (NRRL-B-2356) was ob-
tained from Dr. W. C. Haynes of the Northern
Marketing and Nutritional Research Division,
Peoria, 111. Three mixed-strain freeze-dried
lactic cultures, Mix-6, -17, and -20 were
obtained commercially (Klenzade Products,
Beloit, Wis.). The two Leuconostoc species,
L.-2 and -5 were isolated from the mixed
strain cultures by surface plating according
to the method of Mayeux et al. (1962), and
were tentatively indentified as L. citrovorum.
Mixed-strain and S. diacetilactis cultures
were maintained in sterile 10% reconstituted
nonfat dry milk (NFDM) by transfer of 1%
inoculum every other day and incubation at
21.5° C. for 24 hr. The Leuconostoc cultures
were maintained in a similar manner in sterile
lactic broth (method of Elliker et al., 1956).
Tryptone glucose agar was used to pour plates
which were incubated at 30° C. for 48 hr.,
except the leuconostocs which were held at
21.5° C. for 72 hr. A 10% aqueous solution
of yeast extract (YE) was prepared and auto-
claved at 121° C. for 15 min.
Inoculation of Whole Egg. Aliquots (40 ml.)
of heat treated WE were dispensed into
screw-top culture tubes and 0.8 ml. of a 24
hr. culture was added. Tubes containing WE
were agitated, plated, then inoculated. The
2% inoculum yielded from 5 x 10* to 3 x
 WHOLE EGG FERMENTATION
107 cells/ml. of WE depending upon the
culture.
pH and Desugarization. The pH of heat
treated WE was adjusted with sterile IN lactic
or IN citric acids. Corrections for dilution
were not made. During growth studies, por-
tions of WE were periodically removed and
pH was determined. Extent of desugarization
was observed simultaneously using Clinistic
reagent strips (Ames Co., Elkhart, Ind.).
Diacetyl Determinations. Diacetyl produc-
tion in milk and WE was determined accord-
ing to Pack et al. (1964). Twenty g. samples
were placed in an ice water bath to arrest
growth of microorganisms and to inhibit
diacetyl reductase activity. Analyses for
diacetyl were performed by placing the tubes
in a 65° C. water bath and purging with
nitrogen gas. Diacetyl was trapped as di-
methylglyoxime in buffered hydroxylamine
and converted to a colored complex by reac-
tion with ferrous sulfate. A standard curve
was prepared using dimethylglyoxime in lieu
of diacetyl. Foaming of WE samples was
prevented by adding 5 drops of Dow Corning
Antifoam B. Recovery of diacetyl by this
method was more than 90% at pH 6.0 and
5.5, and was linear with increasing diacetyl
concentrations. Recovery decreased to 87.3%
at pH 5.0. Apparently, diacetyl was trapped
due to coagulation of the WE. At pH 6.5,
recovery was 81.2%, and at pH 7.25, recovery
was only 71.0%.
Sensory Evaluation. Pasteurized WE was
acidified to pH 6.0 with citric acid, inoculated
with S. diacetilactis, then incubated at 21.5°
C. for 8.5 hr. and/or 11.0 hr. Fermented
WE was used to formulate three products:
custard, shortened cake, and a plain cooked
WE spread. The custard and shortened cake
were prepared according to Lowe (1955),
using WE fermented for 8.5 hr., and a fresh
WE control. The cooked WE spread was
1577
prepared by placing fermented WE, incubat-
ed for 8.5 and 11.0 hr., into a plastic pouch
and heating four min. in a 93° C. water bath.
The coagulated WE was cooled and blended
at low speed to a uniform consistency in a
Waring Blendor.
The custards and shortened cakes were
evaluated by 23 panelists and cooked WE
spread by 25 panelists (faculty, staff, and
students). Panelists were served two coded
samples of custard and then two coded sam-
ples of cake: one made with fresh WE and
'one made with fermented WE. Each sample
was scored on a six-point scale ranging from
desirable to undesirable. The cooked WE
spread was spread on plain, unsalted
crackers, and was evaluated on a five-point
scale. Two samples were evaluated at one
time, one sample representing each time of
fermentation.
EXPERIMENTS AND RESULTS
Growth in Pasteurized Whole Egg. Growth
patterns for each culture in pasteurized (60°
C. for 3.5 min.) WE at pH 7.25 are shown
in Fig. 2. Numbers of viable cells in all
FIG. 2. Growth curves of lactic cultures in nor-
mal pasteurized whole egg at pH 7.25 and incubated
at 21.5° C.
1578 S . J . GALLUZZO, O . J . COTTERILL AND R . T . MARSHALL

1 1 1 1
. cultures decreased rapidly during incubation
at 21.5° C. However, S. diacetilactis
Whole Egg (YE)
appeared to recover and grow. Low levels
60° C. - 3.5 min.

,i
pH 7.25 of contaminating organisms, particularly
10 6 heat-resistant bacilli, were difficult to control.
Lack of competition from lactic cultures

,i
enabled these organisms to grow and to
eventually coagulate WE.
S. dia^^Q The addition of yeast extract (YE) to pas-
-
•Si L^S—r* teurized WE did not improve growth (Fig.
3). However, yeast extract (0.1%) was rou-
* ^ » ^ Mix-6 . .*
M i x - 1 7 \ XMix-20
•

tinely included in WE for additional studies

I SSn 1 1 1 since it stimulated rapid acid production and
0 6 12 18 24
desugarization of WE by S. diacetilactis when
Time of Fermentation (hr.)
growth conditions were more favorable.
Lowering the pH to 5.5, 6.0, and 6.5 with
FIG. 3. Growth curves of lactic cultures in nor- lactic acid prior to inoculation permitted S.
mal pasteurized whole egg at pH 7.25 with 0.1%
diacetilactis to grow (Fig. 4). Desugarization
yeast extract added and incubated at 21.5° C.
of the WE at pH 6.0 and 6.5 was completed
within 8-10 hr. At pH 5.5 WE was not
—i 1 I r sugar-free even after 24 hr. The L.-2 and
-5 cultures grew at pH 5.5 in WE. Their
numbers remained static when pH was 6.0
and decreased when pH was 6.5. Initially,
\^-*.°..o_l---~'' numbers of the Mix-6, -17, and -20 cultures

1 1 1 1 _
—i 1 r S3. dia. 20 min.
10 9
- C"' ^-_ti ^^0"*^' 15 min. —o -
-

1(17 i Whole Egg (YE)

65° C.
pH 7.25
•
,

r- 10 5 10 min. ^f^"^
. 1
1 ,

i ^ ^ 5 min.
h

,
\v ... 10 3 I I 1
6.s\ ** *•*""
El L_ 0 6 12 18 24
Time of Fermentation (hr.)

FIG. 4. Growth curves of lactic cultures in nor- FIG. 5. Growth curves of S. diacetilactis in
mal pasteurized whole egg adjusted to different whole egg heated at 65° C. for different times and
pH levels with lactic acid and incubated at 21.5° C. incubated at 21.5° C.
 WHOLE EGG FERMENTATION
***.
Time of Fermentation ,hr.
FIG. 6. Growth curves of lactic cultures in
superheated whole egg adjusted to different pH
levels with citric acid and incubated at 21.5° C.
decreased in WE at all three pH levels.
However, the Mix-6 and -20 cultures grew
slightly after 24 hr. of incubation in WE
adjusted to pH 5.5 and 6.0. Complete desug-
arization was not accomplished by the Leu-
conostoc or mixed strain cultures.
Growth in Superheated Whole Egg. Several
experiments were conducted to determine if
additional heating of the WE would enhance
growth after inoculation. Since S. diacetilac-
tis grew well and showed fermentative prom-
ise, it was used to establish a time-tempera-
ture condition which would permit growth.
Samples of WE (pH 7.25) were heated at
65° C. for 5, 10, 15 and 20 min., cooled, and
then inoculated. Survival or growth of S.
diacetilactis was not improved by heating WE
at pH 7.25 for 5 and 10 min. (Fig. 5). However,
1579
L--S
— ^-
1
1
1 1 ' i -
12 18 24 0 G 12
T i m e of F e r m e n t a t i o n ( h r . )
FIG. 7. Change in pH of superheated whole egg
adjusted to different pH levels with citric acid and
inoculated with the lactic cultures. Incubated at
21.5° C.
heating for 15 and 20 min. permitted growth.
Growth was accompanied by production of
a metallic off-odor which increased with time
of fermentation. A 20 min. time of heating
was used in further experiments. This condi-
tion resulted in a superheated product which
was more viscous but remained liquid.
Subsequently, growth of all cultures was
determined in superheated WE adjusted to
pH 6.0, 6.5, and 7.25 before inoculation
(Fig.6). Citric acid was used for pH adjust-
ment instead of lactic acid because it is a
substrate for diacetyl synthesis. Growth
curves for the six cultures in this series at
pH 5.5 were omitted, since all cultures grew
in superheated WE adjusted to pH 6.0 and
6.5. Only S. diacetilactis and the Mix-20
cultures grew at pH 7.25. However, growth
was more rapid when pH was lower initially.
Mixed cultures produced buttery and / o r me-
tallic odors. S. diacetilactis produced a pro-
nounced flavor change. The problem of con-
taminating bacilli was reduced at pH 6.0.
1580 S. J. GALLUZZO, O. J. COTTERILL AND R. T . MARSHALL

pH Changes in Whole Egg. Changes in pH than for S. diacetilactis, were attained later
resulting from growth of all six cultures in than8 hr. Mix-17reached a peak of 2.3 p.p.m.
superheated WE after adjustment to different diacetyl after 35 hr., and Mix-20 reached 1.5
pH values with citric acid are shown in Figure p.p.m. after 20 hr. incubation at 21.5° C.
7. The mixed cultures, containing acid-
producing streptococci, decreased pH from Production of Diacetyl in Whole Egg.
initial values of 6.5 and 6.0 to pH 5.5 and Studies involving production of diacetyl in
5.0, respectively. L.-2 and -5 cultures, which WE were restricted to samples acidified with
normally produce little acid, caused only citric acid since experiments had shown that
small decreases in pH. S. diacetilactis, on S. diacetilactis did not produce diacetyl in
the other hand, did not decrease pH as much WE acidified with lactic acid. The mixed
as when lactic acid was the acidifying agent cultures produced small and variable quanti-
since it used the citric acid for diacetyl ties of diacetyl in superheated WE. When
synthesis. In a separate experiment, WE the initial pH was 6.0, Mix-6 produced a weak
acidified to pH 5.5 with citric acid (not buttery flavor with 0.5 p.p.m. diacetyl at 12
shown), was desugared by S. diacetilactis hr. and 0.7 p.p.m. at 24 hr. Mix-17 (pH 6.0)
within 10 hr., with a final pH of 5.25. produced a similar flavor, with 0.2 p.p.m.
However, with lactic acid, S. diacetilactis diacetyl at 12 hr. and 0.4 p.p.m. at 24 hr.
lowered pH from 5.5 to pH 4.75, without Mix-20 did not produce diacetyl over the 24
completely desugaring WE. hr. period, but produced the most metallic
off-odor of the three mixed cultures. This
Production of Diacetyl in 10% Nonfat Dry metallic odor was quite volatile since it was
Milk. Amounts of diacetyl produced in 10% rapidly removed from WE during N 2 extrac-
NFDM by the three mixed cultures and S. tion. Mix-6 produced a weak diacetyl aroma
diacetilactis are shown in Figure 8. Diacetyl at pH 6.5 but was not further investigated.
production began immediately in the case of Leuconostoc cultures produced a weak fer-
S. diacetilactis, and peaked at 1.3 p.p.m. in mented odor at pH 6.5 and 6.0. The aroma
8 hr. Mixed cultures, which are dependent was similar to that of a weak solution of
on the action of two types of organisms, began
producing diacetyl only after pH had de-
creased considerably. Consequently, peak Whole Egg (YE)
60° C. - 3.5 min
Whole Egg (YE)
65" C. - 20 min.
concentrations, although they were higher
:
/
'O I
A
/ P t •
10% Nonfat Dry Milk
/ j!«.o \cl

- X^Mix-17 ^fc _
I, \
Mix-20
S. dia. y O v y ^ * / ' v
/ >'
Mix-6
/ /*>—- r\/ <P
/ /
I' VJ.S • • • '
9 12 0 3 6 9 12
Time of Fermentation (hr.)
s of Fermentation (hr.)

FIG. 9. Production of diacetyl by S. diacetilactis

FIG. 8. Production of diacetyl by Mix-6, Mix-17, in normal pasteurized and superheated whole egg
Mix-20, and S. diacetilactis in 10% nonfat dry milk adjusted to different pH with citric acid and incu-
incubated at 21.5° C. bated at 21.5° C.
 WHOLE EGG FERMENTATION 1581

I 1 i i 6 hr. and the metallic odor also was noticeable

6 -" s dia. - at an earlier time.
Whole Egg (YE)
65° C. - 20 min.

f
• ^ \

\ pH 6.0 Sensory Evaluation. According to the t-test

\ (Snedecor and Cochran, 1967), the control
1 \
1
1 <LO
—
— 1 "-•<X
"0-.
s 1
s basis of aroma alone. However there was
o'
^ no statistically significant difference between
<r-
1 i i i
3 6 9 12
Time of Fermentation (hr.)
FIG. 10. Production of diacetyl by S. diacetilac-
tis in superheated whole egg adjusted to pH 6.0
with citric acid and incubated at 30° C.
acetic acid and ethanol. There was no metallic
odor or diacetyl produced by either culture.
S. diacetilactis produced large and equal
amounts of diacetyl in both pasteurized and
superheated WE adjusted to different pH
values with citric acid and incubated at 21.5°
C. (Fig. 9). Peak concentrations of diacetyl
were progressively larger as the initial pH
was decreased from 6.5 to 5.5 with citric
acid. The aroma at pH 5.5 and 6.0 was strong,
resembling that of cultured buttermilk. The
diacetyl peak occurred at 8-10 hr., with the
greatest decrease in its concentration occur-
ring at pH 5.5. The metallic odor which was
present when lactic acid was the acidifying
agent, was evident after the partial reduction
of diacetyl, and was pronounced after 24 hr.
Diacetyl production by S. diacetilactis in
superheated WE at 30° C. was briefly sur-
veyed. The organism was inoculated into
superheated WE acidified to pH 6.0 with citric
acid. Maximal production of diacetyl oc-
curred earlier but was less than at 21.5° C.
(Fig. 10). Desugarization was complete within
custard (mean = 5.2) was significantly (P £
0.01) more desirable than custard prepared
with fermented WE (mean = 3.7). The two
samples were readily differentiated on the
the shortened cake prepared with normal WE
(mean = 5.4) and fermented WE (mean =
5.0). Also, there was no significant difference
between the two cooked WE spreads
prepared with WE fermented for 8.5 hr. and
11.0 hr. The mean scores were 3.2 and 3.1,
respectively.
DISCUSSION
The rapid reduction in numbers of the lactic
streptococci inoculated into pasteurized WE
(pH 7.25) could have been due to biologically
active proteins in egg white (EW), particularly
lyzozyme. It is active against many Gram-
positive microorganisms, including lactic
streptococci (Brown et al., 1962). Lytic activ-
ity is inhibited by yolk, but sufficient activity
probably remained in WE to inhibit lysozyme
sensitive organisms (Cunningham and Cot-
terhill, 1971; and Galyean et al, 1972). The
lesser reduction in numbers and improved
growth at lower pH in the present experiments
can be explained in terms of decreased lytic
activity at low pH (Galyean et al., 1972).
Similarly, superheating WE would reduce
lytic activity and permit better growth (Seide-
man et al., 1963; and Cunningham and Line-
weaver, 1967).
The decrease in numbers and /or poor
growth in pasteurized WE (pH 7.25) may also
have been related to nutrient-binding proteins
in EW. The proteins conalbumin, avidin, and
flavor-protein bind iron, biotin, and ribofla-
vin, respectively. However, WE would
appear to contain sufficient nutrients to satis-
1582 S. J. GALLUZZO, O. J. COTTERILL AND R. T. MARSHALL
fy growth requirements of these organisms,
particularly when yeast extract was added.
S. diacetilactis failed to completely desugar
WE (pH 5.5) in 24 hr. when lactic acid was
the acidifying agent, as low pH apparently
became inhibitory and rate of sugar utilization
slowed. However, when citric acid was the
acidifying agent, S. diacetilactis did not de-
crease pH (even in WE at pH 5.5) as much
because of the utilization of citric acid for
diacetyl synthesis. S. diacetilactis possesses
the enzyme citritase (Harvey and Collins,
1961) that breaks down citric acid to oxalace-
tic and acetic acids. These weaker acids are
further metabolized, causing an increase in
pH which partly counterbalances the de-
crease in pH caused by glucose utilization.
Data on diacetyl production provide an
opportunity to compare single and mixed
culture performance in a model system like
milk and an atypical system like WE. Mixed
cultures contain both acid-producing strepto-
cocci and aroma-producing leuconostocs.
Streptococci lower pH to a level where leu-
conostocs are able to form diacetyl rapidly.
Mixed cultures, therefore, have a certain lag
time prior to diacetyl production, whereas
S. diacetilactis produces both acid and aroma
simultaneously. However, mixed cultures are
preferred over pure cultures for buttermilk
production as excess numbers of S. lactis,
S. cremoris, or S. diacetilactis with relation
to the L. citrovorum population may result
in a "green" (acetaldehyde) flavor (Lindsay
et al., 1965). In this study, diacetyl con-
centrations recorded in milk were within the
expected range for buttermilk cultures. The
decrease in diacetyl levels with time of fer-
mentation is explained by Seitz et al. (1963)
who reported that strains of L. citrovorum,
L. dextranicum, and S. diacetilactis contained
the enzyme, diacetyl reductase, which cata-
lyzes irreversible reduction of diacetyl to
acetoin. This enzyme functions slowly at low
pH.
In contrast, S. diacetilactis performance
in WE was superior to that of the mixed
cultures. S. diacetilactis synthesized diacetyl
so rapidly in WE at pH 5.5 that the zero
hour reading was almost 1 p.p.m., even with
prompt cooling to arrest growth. This rapid
production of diacetyl was probably due to
pre-existing enzymes and high permease ac-
tivity. Synthesis was somewhat slower at pH
6.5 due to lower citrate permease activity,
since this enzyme is most active at lower
pH (Harvey and Collins, 1962). The lower
peak level was probably due to lower citrate
concentration, as Harvey and Collins (1963)
showed that the rate of citrate entry is not
under metabolic control, and is thus not a
function of growth rate. Under given condi-
tions of pH, temperature, and citrate con-
centration, it is a function only of the mass
of cells. The poor diacetyl production by the
mixed cultures may have been partly a result
of the limited sugar supply in WE. Also, only
a small proportion of the cells in mixed
cultures has the capacity to produce diacetyl,
whereas, all the S. diacetilactis cells can do
so. Heterofermentative streptococci are
apparently unable to utilize citrate as a sole
energy source, and need sugar for citrate
utilization (Harvey and Collins, 1963). The
homofermenters of the mixed cultures may
have utilized all the glucose before the leu-
conostocs could become established, thereby
upsetting the normal balance of organisms
in the mixed culture. The two pure-strain
Leuconostoc cultures did not produce diace-
tyl in WE, although, in this case, the sugar
supply was present for diacetyl synthesis.
However, active leuconostocs do not gener-
ally produce diacetyl unless they cannot
complete their metabolic processes or are in
media with a low pH (Keenan and Bills, 1968).
The effect of increasing temperature of
fermentation from 21.5 to 30.0° C. on diacetyl
production by S. diacetilactis was similar to
that reported by Pack et al. (1968) for mixed
cultures. As expected, the culture grew faster
and the diacetyl peak occurred at 30.0° C.
 W H O L E E G G FERMENTATION
The rapid desugarization of WE at 30° C.
and early attainment of the diacetyl peak
offers promise for the development of a rapid,
high yield method of diacetyl production.
Fermentation at a low pH appears desirable
since contaminants were suppressed.
A potential drawback of the use of lactic
cultures is the production of off-odors to-
gether with diacetyl. A strong metallic odor
was noted at times, which suggests the pres-
ence of acetaldehyde. Strains of S. cremoris,
S. lactis, and S. diacetilactis produce signifi-
cant quantities of acetaldehyde (Keenan et
al., 1966). Absence of this odor in the two
cultures of Leuconostoc was expected since
these organisms normally produce little acet-
aldehyde.
The two major features of fermented prod-
ucts: acid and aroma, were apparently impor-
tant in the sensory tests and dictate the
potential usage of fermented WE. High acidi-
ty in fermented WE results in a mealy tex-
tured cooked product. This mealiness was
used to advantage in the preparation of
cooked WE spread. However, the strong
aroma of diacetyl appeared to be either lost
or masked by cooking. Thus, both fermented
and unfermented products were desirable,
and fermentation time was not critical to this
response. The delicate nature of the diacetyl
aroma was further shown by failure of the
panel to differentiate between the control and
cakes containing fermented WE. Aroma and
acidity of fermented WE were masked by
other ingredients or lost in cooking. There-
fore, fermented WE would need to be a
primary ingredient for the flavor to be detect-
ed.
The significantly lower desirability of the
custard prepared from fermented WE may
have been due in part to bias associated with
preconceived ideas about fermented eggs.
Several judges were aware of the nature of
the product and expressed dislike. Terms
used to describe the fermented custard were
predominantly acid and sour. A common
1583
complaint was that this flavor was foreign
to custard.
From observations of the authors, the fol-
lowing considerations emerge. First, the fer-
mented product is acidic and will have limited
application, as do most fermented products.
Secondly, the diacetyl aroma is easily lost
by cooking or is masked by other ingredients.
However, fermented WE has some charac-
teristics not found in normal fermented dairy
products, mainly the emulsifying and coag-
ulative properties of WE. These properties
may be used to advantage in the preparation
of yogurt-like products from WE. The utili-
zation of fermented WE in existing products
may supplement their nutritional value.
ACKNOWLEDGEMENTS
The authors express their appreciation to
Dr. Dee M. Graham for helpful suggestions
and to Dr. Ruth E. Baldwin and her class
in Sensory Analysis for the evaluation of
products.
REFERENCES
Ayres, J. C , 1958. Methods of depleting glucose from
egg albumen before drying. Food Technol. 12(4):
186-189.
Bean, M. L., K. Ijichi, T. F. Sugihara, J. J. Meehan
and L. Kline, 1963. Effects of modified processing
techniques on the performance of egg white solids.
Cereal Sci. Today, 8: 127.
Brown, W. C , W. E. Sandine and P. R. Elliker, 1962.
Lysis of lactic acid bacteria by lysozyme and ethy-
lenediamine tetraacetic acid. J. Bacteriol. 83: 697-
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Cotterill, O. J., 1968. Equivalent pasteurization tem-
peratures to kill salmonellae in liquid egg white at
various pH levels. Poultry Sci. 47: 354-365.
Cunningham, F. E., and O. J. Cotterill, 1971. The
influence of yolk on egg white lysozyme. Poultry
Sci. 50: 1013-1016.
Cunningham, F. E., and H. Lineweaver, 1967. Stabi-
lization of egg white proteins to pasteurizing tem-
peratures above 60° C. Food Technol. 19(9): 136-
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Elliker, P. R., A. W. Anderson and G. Hannesson,
1956. An agar culture medium for lactic acid
streptococci and lactobacilli. J. Dairy Sci. 39: 1611-
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1584 S. J. GALLUZZO, O. J. COTTERILL AND R. T. MARSHALL
Galyean, R. D., O. J. Cotterill and F. E. Cunningham,
1972. Yolk inhibition of lysozyme activity in egg
white. Poultry Sci. 51: 1346-1353.
Harvey, R. J., and E. B. Collins, 1961. Role of citritase
in acetoin formation by Streptococcus diacetilactis
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Keenan, T. W., R. C. Lindsay, M. E. Morgan and
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NEWS AND NOTES
(Continued from page 1554)
The opening Plenary Session is slated for Monday,
August 12, at 10 a.m. in the Municipal Auditorium.
The Rivergate also will be the site for a U.S. A.-Japan
Mycoplasmosis Panel beginning at 1 p.m., a Symposia
Session on Physiology and Management at 2 p.m.,
and concurrent Contributed Paper Sessions, also at
2 p.m.
At 9 a.m. on Tuesday, August 13, a Symposium
on the Economics of Production will be held, followed
at 10.30 a.m. by one on the Economics of Marketing.
Concurrent Contributed Papers also will be presented
that morning.
The first of two special three-hour "emphasis morn-
ings" for pathologists will be held on Tuesday from
9 a.m. until noon. The emphasis will be on diagnostic
procedures for poultry diseases. Under the chairman-
ship of Dr. T. D. Njaka of the West Virginia Depart-
ment of Agriculture, the latest diagnostic methods will
(Continued on page 1591)
Lowe, B., 1955. Experimental Cookery. John Wiley
and Sons, Inc., New York.
Mayeux, J. V., W. E. Sandine and P. R. Elliker,
1962. A selective medium for detecting Leuconostoc
organisms in mixed-strain starter cultures. J. Dairy
Sci. 45: 655-656.
Pack, M. Y., W. E. Sandine, P. R. Elliker, E. A.
Day and R. C. Lindsay, 1964. Owades and Jakovac
method of diacetyl determination in mixed-strain
starters. J. Dairy Sci. 47: 981-986.
Pack, M. Y., E. R. Vedamuthu, W. E. Sandine, P.
R. Elliker and H. Leesment, 1968. Effect of temper-
ature on growth and diacetyl production by aroma
bacteria in single and mixed-strain lactic cultures.
J. Dairy Sci. 51: 339-344.
Seideman, W. E., O. J. Cotterill and E. M. Funk,
1963. Factors affecting heat coagulation of egg
white. Poultry Sci. 42: 406-417.
Seitz, E. W., W. E. Sandine, P. R. Elliker and E.
A. Day, 1963. Distribution of diacetyl reductase
among bacteria. J. Dairy Sci. 46: 186-189.
Snedecor, G. W., and W. G. Cochran, 1967. Statistical
Methods, 6th Edition. The Iowa State University
Press, Ames, Iowa.
Stewart, G. F., L. R. Best and B. Lowe, 1943. A
study of some factors affecting the storage changes
in spray-dried egg products. Proc. Inst. Food Tech-
nol., pp. 77-89.
be discussed and demonstrated by experts from the
United States and overseas.
On Tuesday afternoon, a Symposium on Nutrition,
and presentations of Contributed Papers are slated.
The U.S.A. Branch of the World's Poultry Science
Association has a General Membership meeting
scheduled for 5 p.m.
A Symposium on the Pigments Involved in Giving
Egg Yolks and Broiler Carcasses the Color Consumers
Like will be held on August 13 with Dr. M. L. Sunde
of the University of Wisconsin as Session Chairman.
Speakers from several foreign countries, as well as
the U.S., will discuss the various materials and
compounds found in nature that will produce the
desired color. Dr. Braeunlich of Basel, Switzerland,
will discuss the chemical characteristics of carotenoids
utilized by chickens, and the methods used to evaluate,
chemically and visually, the efficiency of a number