LINEAR PROGRAMMING EVALUATION OF FEEDS 107

on livestock feeds through parametric linear
programming. Cereal Science Today, 16(6):
181-189.
Frazer, J. R., 1968. Applied Linear Programming.
Prentice-Hall, Inc., Englewood Cliffs, N.J.
Harvey, D., 1970. Tables of the Amino Acids in
Foods and Feedingstuffs. Technical Communi-
cation No. 19. Commonwealth Agricultural Bu-
reaux, Farnham Royal, Bucks, England.
Institute of Food and Agricultural Sciences at the
University of Florida, 1969. Proceedings of a
Seminar: An evaluation of animal nutrition
data used in the computer formulation of ra-
tions. Gainesville, Fla.
National Research Council, 1971. Nutrient Re-
quirements of Poultry. National Academy of
Sciences Printing and Publishing Office, Wash-
ington, D.C.
Parkhurst, R. T., 1967. Commercial Broiler Pro-
duction. Agricultural Handbook 320. Govern-
ment Printing Office, Washington, D.C.
Stafford, L. H., L. E. Ott and L. C. Snyder, 1965.
Managerial aspects of least-cost feed formula-
tion with linear programming. Marketing Re-
search Report No. 729. Government Printing
Office, Washington, D.C.
Taylor, R. D., G. O. Kohler, K. H. Maddy and
R. V. Enochian, 1968. Alfalfa meal in poultry
feeds—an economic evaluation using parametric
linear programming. Agricultural Economic Re-
port No. 130. Government Printing Office,
Washington, D.C.

Pickled Eggs
1. P H , RATE OF ACID PENETRATION INTO EGG COMPONENTS
AND BACTERIOLOGICAL ANALYSES1

J. C. ACTON AND M. G. JOHNSON

Department of Food Science, Clemson University, Clemson, South Carolina 29631

(Received for publication April 7, 1972)

ABSTRACT The white and yolk components of hard cooked eggs pickled in 3 % or
5% acetic acid vinegars reached nearly equal pH values of 4.0-4.1 within six days after
preparation. The pH declined rapidly in the egg white with a slower decrease occurring
in the yolk. Maximal acetic acid concentrations were attained in the white in one day.
Complete penetration of acid through the white to the yolk required 3 to 6 days to reach
an equilibrium concentration and was dependent on the initial acid strength of the pickling
solution. From day 6 to day 15, only slight changes in pH or acid concentration of the
pickling solution, white or yolk were noted. Pickling spice ingredients reduced the initial
acid strength of the pickling media ( 3 % or 5%) by 20-23%. Bacteriological analyses of
pickling solutions indicate that the pickled eggs were bacteriologically safe.
POULTRY SCIENCE 52: 107-111, 1973

INTRODUCTION conditions, pH of the egg albumen and cook-

OTUDIES of hard cooked eggs have
^ been conducted to evaluate the corre-
lation of initial egg quality and method of
cooking with peeling problems (Swanson,
1959; Meehan et al., 1961; Reinke and
Spencer, 1964; Fuller and Angus, 1959;
Irmiter et al., 1970). In general, storage
1
Technical Contribution No. 1001 of the South
Carolina Agricultural Experiment Station, Clem-
son University, Clemson, S.C. 29631.
ing method were directly related to the
cooked egg's peelability.
Hard cooked eggs when packaged exude
liquid with a greenish yellow tint. Chemi-
cal analysis of exudate (Nath et al., 1971)
showed components which appeared to
originate solely from the egg white.
Baker et al. (1967) examined factors af-
fecting the discoloration of hard cooked egg
yolk. The yolk pH, high cooking tempera-
ture, long cooking time and long storage
108 J. C. ACTON AND M. G. JOHNSON
before cooking significantly influenced the
extent of yolk discoloration.
The demand for hard cooked eggs ap-
pears to be increasing in the food service
industry (Lauser, 1971). Several pro-
cessors are marketing a portion of the hard
cooked egg volume as pickled eggs. Cun-
ningham et al. (1971) have prepared and
tested pickling formulas primarily for home
conditions.
The purposes of this study were to deter-
mine: (1) the changes of pH in pickling
solutions and in hard cooked eggs during
pickling and (2) the rate of acid penetra-
tion into the white and yolk of hard cooked
eggs. An analysis of the pickling solution
for bacteria of public health significance was
included in this study.
EXPERIMENTAL
Preparation of hard-boiled eggs. Fresh
Grade A small eggs were obtained from a
local egg packing plant and held in a 10°C.
room for 2 to 8 weeks. As needed, the eggs
were removed from storage and allowed to
equilibrate at room temperature (22-24°
C.). Batches of two dozen eggs were placed
in a boiler, covered with 22-24°C. water
and placed on a cool heating unit. The wa-
ter temperature was raised to 85 to 88°C.
(in approximately 20 min.); the eggs were
held in this temperature range for 25 min.
At the end of the heating period, the eggs
were rapidly cooled to room temperature
with an ice-water bath. The eggs were
hand peeled, rinsed free of shell and mem-
brane fragments and stored under distilled
water. No eggs were used which exhibited
tear damage from peeling.
Pickling. Stock distilled vinegar contain-
ing 12% acetic acid (120 grain) was di-
luted with distilled water to prepare vine-
gar solutions of either 3 % or 5% acetic
acid (w./v.). The pickling ingredients con-
sisted of 40% (w./v.) sucrose (for 3 %
acetic acid) or 45% (w./v.) sucrose (for
5% acetic acid) and 6% (w./v.) commer-
cial pickling spice.2 These were added to
heated vinegar solution at 80°C. The pickle
solution was further heated to 85°C. with
continuous agitation using a stirrer-hot
plate (Corning PC-351).
Two eggs per half-pint jar were covered
with the hot pickle solution. Approximately
10 g. of spice mixture was included in each
jar. The jars were sealed and held at room
temperature (22-24°C.) until analyzed for
pH and acidity.
pH and acidity. The pH of freshly pre-
pared pickle solution and of separated
white and yolk from 6 fresh hard-boiled
eggs was determined with a pH meter. Yolk
and white samples were blended with 5X
their weight of distilled water for pH mea-
surements. Duplicate samples of the sus-
pension were used. These initial pH values
of the white and of the yolk were used as
the titration end-points in measuring acid-
ity during pickling intervals.
Samples were analyzed for pH and acid-
ity at the following periods: 2, 4, 6, 9, and
12 hours and 1, 2, 3, 6, 9, 12 and 15 days.
The contents of duplicate jars at each time
interval were removed and separated into
solution, white, and yolk. Acidity, expressed
as percent acetic acid, was determined by
titration with 0.1 N NaOH. Samples sizes
were as follows: two-5 ml aliquots of pickle
solution; two-50 ml. aliquots of yolk or of
white suspension. The 50 ml. samples rep-
resented 10 g. of original yolk or white.
Plate counts. Pickling solution samples
of the 3% acetic acid were taken asepti-
cally from freshly opened jars of eggs and
decimal serial dilutions prepared with ster-
ile saline. Appropriate dilutions were
poured in duplicate with the following agar
2
Commercial (C. F. Sauer Co.) pickling spice
ingredients: mustard, coriander, cinnamon, gin-
ger, allspice, dill seed, black pepper, bay leaves,
cloves, chillies and mace.
 PICKLED EGGS 109

media: total plate count agar (Difco), for

total viable count; Levine's Eosin Methy-
LEGEND:
lene Blue agar (Difco) for Escherichia and X X SOLUTION

Enterobacter; Staphylococcus 110 (Difco), •

A
•
A
YOLK
WHITE

for staphylococci and micrococci (surface

plating technique used). Total aerobic and
anaerobic spore counts were determined I 6.0 -
using pickling solution heated at 80°C. for
15 min. After samples were heated, decimal
dilutions were poured in duplicate with
Trypticase Soy Agar (Bioquest Labora-
tories), for total aerobic counts. Total
counts of anaerobic spores were determined -1 1 1 1 L,A_J 1 l „ _ i -

by pouring samples with Fluid Thioglyco- 0 2 A 6 8 10 12 I 2 3 6 9 12 15

i HOURS " DAYS •"

late Broth (Difco) containing 1.5% agar.
After the latter plates hardened, they were FIG. 1. The pH of pickle solution, egg yolk
and egg white from initial 3 % acetic acid vinegar
sealed in an anaerobic atmosphere provided as a function of time.
by the Gas Pak system (Bioquest). All
plates were incubated for 48-72 hr. at 30°C. riod after pickling of hard cooked eggs in
Statistical analyses. Results were sub- 3% acetic acid vinegar are shown in Figure
jected to analysis of variance and the sig- 1. The pH of the pickle solution gradually
nificance of means tested by Duncan's shifted during the first day from an initial
method (Steel and Torrie, 1960). value of 3.4 to a final pH of 4.2 or 4.3 in
2-4 hours. The yolk pH decreased more
RESULTS AND DISCUSSION slowly requiring 6 days to reach the same
The changes in pH of pickle solution, pH as the pickle solution. The trends for
egg white and egg yolk during a 15 day pe- pH changes in eggs pickled in 5% acetic

TABLE 1.—Percent acetic acid in pickle solution1 and egg components2 from 3% and 5% acetic acid vinegars'

3 % HOAC Vinegar 5% HOAC Vinegar

lime Egg Component Egg Component
urs-Days Pickle Pickle
Solution Solution
White Yolk White Yolk

0- 0 2.30" 0.00" O.OO 4.02" 0.00" 0.00"

2- 0 1.71 b 0.70° 0.09 b 2.94 b l.llb 0.14°
4- 0 1.58° 0.86° 0.13° 2.79b° 1.28° 0.30°
6- 0 1.54°d 0.96 d 0.26° 2.72 bcd 1.48d o^S"*1
9- 0 1.50"1 1.06° 0.36 d 2.69M 1.64d° 0.46 d
12- 0 1.46d 1.07«' 0.40* 2.49cde 1.61 d 0.68°
1 1.29s 1.12°* 0.50° 2.48«ie 1.80° 1.10?
2 1.14* 1.12°* 0.72 f 2.48°d° 2.14* 1.40s
3 1.18f 1.17« 0.80« 2.48 ede 2.26* 1.60"
6 1.11* 1.18* 0.91 h 2.43°d° 2.22* 1.74;i
9 1.05? 1.12°* 0.88 h 2.21° 1.99' 1.53 h
12 1.11* 1.16* 0.88 h 2.36d« 2.14* 1.69"
15 1.13* 1.17« 0.93" 2.21« 2.04 f 1.58"
1
Percent acetic acid (w./v.).
2
Percent acetic acid (w./w.).
3
Any two means within a column having one or more of the same letters are not significantly different
at p<0.05.
110 J. C. ACTON AND M. G. JOHNSON

TABLE 2.—Change of acetic acid concentrationegg white, and (2) initial acid concentra-
during pickle solution preparation1
tion in the pickling medium. The decrease
Stage of Preparation Percent HOAC of acid strength of the pickle solutions in
contact with eggs (Table 1) resulted from
Initial vinegar solution 3.02
Heated to 80°C. 3.02 the acid absorption by both the egg white
Pickle ingredients added, 2.32 and yolk.
heated to 85°C.
During preparation of the pickle solution
Initial vinegar solution 5.02 and prior to addition of the eggs, a signifi-
Heated to 80°C. 5.00
Pickle ingredients added, 4.02 cant reduction of the calculated acetic acid
heated to 85°C. concentration occurred (Table 2). Sam-
1
Averages for 3 preparations. pling the solution at various stages of prep-
aration showed a 20-23% decrease of acid
strength, probably due to the absorption
acid vinegar were parallel to those shown in and neutralization of acid by the pickling
Figure 1. In many cases, final pH values spice ingredients. This reduction of acid
were equal to those obtained with the 3 % must be considered when attempting to reg-
acetic acid vinegar. ulate the final concentration in the pickled
The percent acetic acid in the pickle so- eggs.
lution and egg components are presented in The level of various bacteria of public
Table 1. In the 3 % acetic acid vinegar, the health significance in pickling solutions of
acid concentration rapidly increased in the the 3 % acetic acid vinegar is shown in Ta-
egg white during the first 9 hr. and fluctu- ble 3. Total viable counts were low and no
ated between 1.12 and 1.18% through the coliforms were recovered. Although some
remainder of the 15 days. The yolk acid staphylococci were found, none appeared to
concentration significantly increased at be coagulase positive based on colony color.
each interval up to day — 6. Similar results Samples at 50 days had nearly the same
were found for eggs pickled in 5% acetic relative bacterial counts as those stored for
acid vinegar although the actual quantities 20 days. The data for spore counts indicate
of acetic acid available for penetration that sporeforming bacteria, probably in-
were greater. The penetration of acid to the troduced with the spices and sugar, survive
yolk is probably dependent on two factors: the harsh environment but do not increase
(1) diffusion rate of acetic acid through the in numbers. These analyses, coupled with

TABLE 3.—Numbers of bacteria from pickling solution {3% acetic acid vinegar) at two storage intervals

Colony forming units 1 per ml of solution on:

Sampling Interval Spore Counts 3

of Total Levine's Staph 110
Pickling Solution Plate Count EMTS Agar2 Trypticase Thioglycollate
Agar Agar Soy Agar Agar
(aerobic) (anaerobic)
20 days 8.5X10 2 5.1X10 2 2.1X10 2 7.5X10 2
50 days 7.5X10 2 6.5X10 2 5.4X10 2 8.0X10 2
1
Values are average of duplicate determinations.
2
No orange or yellow pigmented colonies were observed, suggesting that none were coagulase positive;
coagulase production was not tested.
' Pickling solution samples heated at 80°C. for 15 min. prior to plating.
 PICKLED EGGS
pH and acidity values for the pickling solu-
tion and eggs indicate that these hard
cooked eggs, pickled as described, were
bacteriologically safe.
Further studies are in progress to evalu-
ate the rate of flavor development and the
change of egg tenderness which occurs dur-
ing the pickling process.
ACKNOWLEDGMENT
The authors thank Miss Patricia Ros-
tron and Mr. Harry Findley for technical
assistance.
REFERENCES
Baker, R. C , J. Darfler and A. Lifshitz. 1967. Fac-
tors affecting the discoloration of hard-cooked
yolks. Poultry Sci. 46: 664-672.
Cunningham, F. E., D. J. Mugler and 'M. E. Jack-
son. 1971. Try pickling at home. Cooperative
NEWS AND NOTES
(Continued from Page 101)
try nutritionist, Dr. Edwards has been a university
faculty member since 19S7. He is Professor of
Poultry Science in the College of Agriculture and
has just completed nine months of study in France
and England on a Guggenheim Fellowship where
he was working on a textbook on the fundamen-
tals of animal nutrition.
For the past ten years, Dr. Edwards held a Re-
search Career Development Award of the National
Institute of Health. In 1962, he received the Amer-
ican Feed Manufacturers' Award of the Poultry
Science Association and was the recipient of the
Senior Faculty Award of the University of Geor-
gia Chapter of Gamma Sigma Delta.
Dr. Edwards received his B.S.A. degree from
Southwestern Louisiana Institute, his M.S. degree
from the University of Florida, and his Ph.D. de-
gree from Cornell University.
Dr. Allan Rahn joined the faculty of the De-
partment of Poultry Science as Assistant Professor,
effective September 1, 1972. He is a native of Illi-
nois and received both the B.S. and M.S. degrees
from Southern Illinois University and the Ph.D.
degree from Iowa State University. He will teach
(Continued on page 126)
111
Extension Service, Kansas State University,
Manhattan, Kansas. Bull. L-227.
Fuller, G. W., and P. Angus. 1969. Peelability of
hard-cooked eggs. Poultry Sci. 48: 1145-1151.
Irmiter, T. F., L. E. Dawson and J. G. Reagan.
1970. Methods of preparing hard cooked eggs.
Poultry Sci. 49: 1232-1236.
Lauser, G. 1971. Processor claims pre-cooked eggs
cheaper for institutional trade. Feedstuffs,
43(31): 6.
Meehan, J. J., T. F. Sugihara and L. Kline. 1961.
Relation between internal egg quality stabiliza-
tion methods and the peeling difficulty. Poultry
Sci. 40: 1430-1431.
Nath, K. R., D. V. Vadehra and R. C. Baker.
1971. The chemical nature of cooked egg exu-
date. Poultry Sci. SO: 1611.
Reinke, W. C , and J. V. Spencer. 1964. Observa-
tion of some egg components in relation to
peeling quality of hard-cooked eggs. Poultry
Sci. 43: 1355.
Swanson, M. H. 1959. Some observations on the
peeling problems of fresh and shell-treated eggs
when hard cooked. Poultry Sci. 38: 1253-1254.
and conduct research in the area of production
economics and marketing.
New graduate students in the Department of
Poultry Science are: W. Calhoun, B.S. (Ashland
College), M.S. (Duquesne University); D. D.
Dykstra, B.S., M.S. (Iowa State University); R. C.
Jackson, B.S.A. (University of Georgia); H. R.
Jordan, B.S., M.S. (Pennsylvania State Univer-
sity) ; T. Murai, B.A., M.A. (Hokkaido Univer-
sity) ; P. L. Potts, B.S.A. (University of Georgia);
E. Simhaee, B.S., M.S. (Pahlavi University); D. R.
Wilburn, B.S. (University of Georgia); C. Wu,
B.S. (Taiwan University).
Dr. K. F. A. Soliman received his Ph.D. degree
at the August graduation and has accepted a posi-
tion as Assistant Professor at Tuskegee Institute.
The following graduate students received mas-
ter's degrees at the June and August graduations:
V. T. Cummings, C. G. Janney, J. L. Mabon,
L. K. Misra, J. W. Page, J. T. Rice, W. E. Sexten,
and Cora C. Yen.
ALABAMA NOTES
The Alabama Feed Association has contributed