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 * UNITED STATES
DEPARTMENT OF AGRICULTURE
LIBRARY

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 ‘™, BULLETIN OF THE
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No. 51

Contribution from the Bureau of Chemistry, Carl L. Alsberg, Chief.

July 20, 1914. -
(PROFESSIONAL PAPER.)

A BACTERIOLOGICAL AND CHEMICAL STUDY OF

COMMERCIAL EGGS IN THE PRODUCING DIS-
TRICTS OF THE CENTRAL WEST.'
Under the direction of M. E. Pennineton, Chief, Food Research Laboratory,
associated -with M. K. JENKINS, E. Q. St. JoHN, and W. B. HICKS.

INTRODUCTION.
The deterioration of eggs between the time of their production on
the farm and their arrival at the consumer’s table has been discussed
from different viewpoints in the publications of the Department of
Agriculture. Among these discussions have been the various causes
contributing to the loss of freshness, the necessity for lessening the
number of eggs that become totally unfit for food each year, the
results of improved methods of handling, and the fixing of respon-
sibility for the enormous economic and financial loss due to decay.
The scientific literature on the bacteriology and chemistry of the
ecg of the domestic fowl is surprisingly meager, considering the
many ages that the egg has served as an important article of food.
The distinction between a good and a bad egg has rested more often
on individual opinion, colored by prejudice or preference, than upon
1 This report embodies a study of eggs, the first of a series of reports from the investi-
gation of frozen and dried eggs, which is being conducted by the Food Research Laboratory
of the Bureau of Chemistry. It deals with the subject of the quality of the eggs which
go to the egg-breaking establishments in the egg-producing sections of the Central West.
It is the basis on which the study of industrial problems and conditions must be founded.
The second report of the series will deal with the commercial procedures in the packing
houses, the environment, and methods of work which must be followed if a high-quality
product is to be produced. Other phases of the investigation will be embodied in reports
on the general management in the packing houses making for efficiency, and on the treat-
ment wuoich the output receives in the establishments of the bakers, who are the chief
users of frozen and dried eggs. For the data submitted we are indebted to M. K. Jenkins,
who has studied a number of individual eggs, prepared the ‘“‘ commercial’’ samples, and
has been of very material assistance in the correlating and presenting of the facts; to
EH. Q. St. John, who has done the greater part of the bacteriological work; and to W. B.
Hicks, who is chiefly responsible for the chemical analyses. The Omaha Food and Drug
Inspection Laboratory, with its force of chemists, in charge of 8S. H. Ross, was assigned
to this investigation with the staff cf the Food Research Laboratory for the summers of
1911 and 1912, and was made the headquarters for the field investigations.
Noty.—This bulletin gives details of an extensive study of commercial eggs and makes
recommendations for improvements in handling. While the study was made in the Central
West, the bulletin is equally. of interest to all sections where eggs are produced in com-
mercial quantities and are sent to egg-breaking or other packing establishments.

17625°-—14—__1
2 BULLETIN dl, U. S. DEPARTMENT OF AGRICULTURE.

the exact knowledge of the composition or history of the egg itself.

The farmer’s wife, accustomed to the fresh eggs of the farm, dis-
cards as unfit for food an egg which the city baker would, with a
clear conscience, serve to his own family. The difference in the point
of view has progressed even to the savants who have held opposite
and positive opinions concerning the wholesomeness or desirability
of certain eggs, especially of those known in commerce as * frozen ”
or “ desiccated.”
Investigation has shown that when the egg is laid it is of a fairly
constant chemical composition and contains but few bacteria or
molds. In the vicissitudes of marketing, with its attendant unde-
sirable conditions, eggs in the shell undergo a variety of changes,
referable, almost exclusively, to the mode of handling. These
changes, their effect upon the food and market value of the egg,
and the means by which they can be reduced to a minimum have
been, and still are, a subject of investigation by the Department of
Agriculture cooperatively with every branch of the egg-marketing
industry.
The great variety of conditions to which an egg is subjected and
its sensibility to temperature, humidity, odors, etc., result in many
evidences of deterioration in the eggs on the market. The extent to
which such downward changes are reflected in the composition of the
egg, together with their recognition by physical, chemical, and bacte-
riologic methods, is the subject of the present report.
The work has been done as part of an investigation of the prepara-
tion of frozen and dried eggs. It is the foundation on which to build
all the other phases of the investigation, such as the construction
of the apartments in which egg breaking for food purposes can pro-
ceed; the methods used to guard against bacterial contamination; the
systematic application by the employees in the packing houses of the
knowledge gained by scientific research to increase accuracy and
efficiency ;the quality of the output of the houses under old and new
conditions, and the behavior of those products when they reach the
hands of the baker, who, for this investigation, may represent the
consumer. These diverse parts of the work will be reported in a
series of publications of which this is the first. It will be observed,
therefore, that the subject is discussed from the industrial viewpoint,
even though it is essentially a laboratory study.
The individual eggs, and some of the composite egg samples, were
opened in the laboratory; the samples designated “ commercial ”
were opened by a bacteriologist in packing houses where the sur-
roundings were as clean as, or cleaner than, in the laboratory and
where all utenstils were sterilized. -
The information given in this report has been gathered in south-
western Iowa, northern Tennessee, and the valley of the Missouri
from the northern border of Iowa to the central part of Kansas.

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 COMMERCIAL EGGS IN THE CENTRAL WEST. 3

The eggs were from sources comparatively close to the investigators;

that is, the haul was seldom more than 200 miles. Had the eggs not
been broken at these first, or, at most, second concentrating centers,
the probability is that they would have been shipped a four to seven
days’ haul before reaching a consuming center. They were, there-
fore, in a correspondingly better condition because broken nearer
their point of origin. The field work reported was carried on dur-
ing the summers of 1911, when exceptionally hot weather prevailed
over an unusually wide territory, and 1912, which was not an un-
usual summer in any respect. The individual eggs, however, were
studied between the winter of 1910 and the autumn of 1912.
To comprehend the egg on the market it is necessary to determine
first the condition of the absolutely fresh egg, that a standard of
comparison may be obtained, and then the condition of the eggs on
the market to see wherein and how much they differ from the fresh
article. It is also highly desirable to observe the character of the
eggs bought: by the housewives at the corresponding time, in the
same locality, to see whether there are any material differences be-
tween the eggs broken for home cookery and those broken by the egg
canner who supplies the public baker.
FRESH EGGS.
BACTERIAL CONTENT.
The chemical and bacteriological characteristics of perfectly fresh
eggs—that is, eggs which are not more than 24 hours old and which
are kept in a cool place—have been given by the first author of this
report in a previous communication, entitled “A Chemical and Bac-
teriological Study of Fresh Eggs.”? In this study 150 high-quality
eggs, not more than 24 hours old, were examined for the bacterial
content in white and yolk. A strictly fresh egg is pictured in Plate I
(see at end of this bulletin). Aseptic precautions were used in ob-
taining samples and all the work was done on the basis of weight,
not volume, since the latter introduces a decided error in so viscous
a substance as egg. A summary of the results shows that there was
found an average of 2 organisms per gram in the white and 6 per
gram in the yolk when the incubation temperature was 37° C., and
7 organisms per gram in the white and 9 per gram in the yolk when
the incubation was at 20° C. It may be said that these eggs were
gathered between February and November, inclusive.
Stiles and Bates,? in a recent study of 616 fresh eggs gathered
between April and October, found that the average infected yolk
contained 271.7 organisms to the cubic centimeter and the infected
white 15.9 organisms. They also found, however, that in 13.99 per
1J. Biol. Chem., 1910, 7 (2): 109.
2A Bacteriological Study of Shell, Frozen and Desiccated Eggs Made Under Laboratory
Conditions. U.S. Dept. Agr., Bureau of Chemistry Bul. 158, 1912.
4 BULLETIN 51, U. S. DEPARTMENT OF AGRICULTURE.

cent of the yolks examined there were no organisms present in 1 cc

of the material, and the white of the egg was sterile in 1 cc quanti- |
ties in 32.18 per cent of the samples examined. Maurer! reported
81.9 per cent of the eggs he examined to be sterile, and stated that
of the 18.1 per cent infected 82 per cent were infected in the yolk,
25.9 in the white, and only 7.9 per cent in both yolk and white.
Maurer used aseptic precautions in obtaining samples of the egg
material examined. Stiles and Bates did not clean the shells, and
their method of opening was to crack on the edge of a sterile Petri
plate and shift the yolk from shell to shell in housewife fashion, to
effect a separation of the two substances. The plate used for crack- .
ing the shell was also the container of the sample. Maurer does
not give the number of organisms occurring, merely stating their
presence or absence.
The bacterial content of fresh eggs has been proved to be widely
diversified in the character of the organisms present, but their num-
bers are small. The varieties of organisms present will be considered —
elsewhere. It may be said, however, that Pennington? did not find
B. coli in any of the 150 eggs examined; neither did Maurer in a
study of 160 eggs, many of which had dirty shells or were placed
under artificial conditions favoring shell penetration. B. coli were
found on the shells in many cases. Stiles and Bates found one egg
Jaid in the month of July that contained B. coli in the yolk. It is
possible, however, that this might have been the result of contamina-
tion while cracking on the edge of the Petri dish or separating white
and yolk by the shell method of the housewife. With the exception
of this one egg yolk wherein B. coli were reported, the examinations
of fresh eggs do not, from a practical industrial viewpoint, show
conflicting testimony. They agree fairly well in asserting that the
fresh, well-handled egg, though not always sterile, is not, on the other
hand, infested by large numbers of bacteria, and &.coli are practically e
ee

never present.
After bad handling or mistreatment the number of organisms e

may, and frequently does, increase enormously. Whether they in-

variably appear in numbers after bad handling or age has inter-
fered with the integrity of some one or more of the component parts
of the egg is a problem to be solved. The studies here chronicled
are expected to throw some hght on this question. Such studies
would indicate also whether the large numbers of bacteria found in
certain eggs are due to a rapid increase of the original organisms
found in them even while still present in the oviduct, or whether an
additional infection through the shell is common in the course of
the usual routine of marketing.
1 Bacteriological Studies on Eggs. Kansas State Agricultural College Bul. 180, 1911.
2J. Biol. Chem., 1910, 7 (2): 109. plies
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 COMMERCIAL EGGS
IN THE CENTRAL WEST. 5
CHEMICAL COMPOSITION.

Numerous chemical analyses of fresh eggs have been made in this

laboratory in connection with the investigation of the handling of
eggs. They extend our knowledge of the composition of the fresh
egg, especially in relation to the quantity of loosely bound nitrogen
in egg protein; that is, the nitrogen split by the action of a weak
alkali and removed for estimation by aeration. This form of ni-
trogen occurs in very minute quantity in the protein of the freshly
laid egg, and is much increased, though still: small in amount, in
eggs that have deteriorated. The amount of loosely bound nitrogen
is at the present time the best and simplest index that has been found
of the chemical stability of the egg. Hence the quantity of this
substance has been determined in the various grades and kinds of
market eggs and in those used by the egg breakers, as well as in eggs
24 hours old.
The analyses recorded in Table 1 show that most of the loosely
bound nitrogen is found in the yolk of the egg, where it averages in
western summer eggs of highest quality 0.0023 per cent. Whole eggs
of like grade gave 0.0013 per cent as an average.
TABLE 1.—Fresh eggs.
|
Total number |
of bacteria per |Gelatin| Ammoniacal |
gram on plain | jique- |nitrogen, Folin
Date of |agar incubated | fying method. : h Size
Portion and sample No. collec- at— organ- vee. Monee
tion. elsms Sead Oba *)/sample,

° or70 ram.
ae Vet
Wet |- D Dry
20°C. |37°C. |8 basis. | basis.
|

| JEG IEG Hach, Neer, R

White: 1911. cent. | cent. | cent. | cent. | Fggs.
26s see ote ware ee Ay ee cegica| heegee NS seeaaerd OF 000441020033" hae casce noe. ae 18
Fal asBIE rc i ABUNias a elts eee Snedecor teehee 4 a eg ae OGC3A) 20025 es <3 5 eee 12
DOTS i as saa ee PAU GR Oi tte Seep es |e esa ealaiee a OOO QOS BHIskeet 12
OOO Mate na sekee ee 224d6 WySEES iris Sau Riad sa aeeee OOO3 4) 200257|= <2 ee). Gs es 12
Dele aes ace Maesee? --do pO ae atya terierearals
Gees eee QO0S cE 400250 was |esee: 12
Yolk }
DG ORB 3 aoe eee Ee July 10 150 | 360 130 0026 | .0050 | 47.81} 31.87 6
PAG SpE eeWARS Sea eyo Aa mamateeSt PU ye RUAN) ANS Esp is eres aie as Bees 0025 | .0047 | 46.88 |..--__.- 18
ZI Seesoislerne a ees ee Sl eri Werte Wael uri Ses Sin ees 0023 | .0043 | 46.89 |........ 12
DORE ee ea a. et AUIS OTN Cs ea Svea se alts ag Mae 2 0024 -0046 | 47.80 |_..-...: 12
BPE eee een eae & do feet ane[e gep iets: Sele a ad ges 0020 |} .0034 | 47.28 |........ 12
NNN 2 = hese cess eee Mos ya Eres eas see Eee ae 0022 | .0042 | 47.84 |__...:-: 12
aihole2 BE:
pp eeRar Brey ce eeeeraS 2 June 26 0 Dee eee ee IE enema eon On 10286 30
ae Sa Sanaa aa oe -do 0 OP EZ SS ARS Se ae Se Dice 73.44 | 11.28 30
NGS ee ee July 3 150 | FOE Saal es ea 72.96 | 11.40 36
DOG) Sia ee og Ee July 19 i) AO le ee Semen OOH S82 NOO4 ies ae eee sea 6
UGE SENSE Sere Leamangtd Sy Aes fe ss Salas hie 0011 QO40) jeccxnee ete a a ee ie
BORER SE a ie RE aD ool es ees ysl meer leer haa Tee 0013 QOSTAI es Gale eae 1h
CURES Reson ress 3eerie adore 7 HA See eee Seema clams etre Mens ies be ere a 6
SAS ee one cea neces eee eee July 24 45 DO eaaen eee - 0015 LUE beleee el Bed ea red
SOU Seer eke acc Seto. Sie 0 85 AD) Beran nies . 0013 OO4G ieee RARE.
LU Sy Se conee oH ee ao bee US ae area gees Joelle c ac eae - 0011 004072 ere soe: 24

1 Moisture content of fresh egg white taken as 87.94 per cent, average of 236 eggs.
2 Moisture content of fresh whole egg taken as 72.44 per cent, average of 9 eggs (analyses given in KOnig,
pp. 98 and 1470, and of three analyses Nos. 126, 127, and 168 given above).

A number of unpublished analyses of fresh eggs from various

sources would indicate a maximum and minimum variation of 0.0010
6 BULLETIN 51, U. S. DEPARTMENT OF AGRICULTURE.

and 0.0015 per cent nitrogen, respectively. The analyses of summer

egos made in the western egg-producing district are comparatively
few, but since their composition proved to be like that of the eggs
studied in the general egg-handling investigation it was not deemed
necessary to multiply them beyond the number needed to confirm
existing data.
The moisture content of the summer egg is of interest. It may be
that there is a relation between the amount of water present in the
egg and its resistance to decay. A study, in another connection, of
perfectly fresh eggs from a well-cared-for flock during different
seasons, when the fowls showed physiological variations, is given in
Table 2.
TABLE 2.—Water content of fresh eggs.

Number Indi- Moisture.

of vidual
peie samples. eggs.
W hite. Yolk.

Per cent. ; Per cent.

WabsesstoiMars 162. 9... feeei\s wees UES AS oe ee eee me | £4 181 87.90 47. 44
[Nae os to Sop 7 eee i Pee Mer ORG te Seems { eee - \ 183 | 88.19 47.96
OCtostORNO We 2s Se Sas a a ae RE eee See ee eae ee 4 | 54 87.99 47.54

Tt will be seen that in August and September, when the industry

considers the quality of the eggs lowest, the maximum quantity of
water is found in both white and yolk. In the early spring, when
eggs are undoubtedly of highest quality, the water content is lowest,
and in the cool days of autumn it occupies a medium position. The
statement of the increased water content of summer eggs is not made
authoritatively, but only as a promising line of investigation on this
subject. It can be emphatically stated, however, that analytical dif-
ferences in tenths of per cent may, in so specialized a tissue as an
ego, carry with them marked variations in physiologic functions and
chemical stability.
The fat of an egg is almost exclusively in the yolk. According to
Pennington? the ether extract of the yolk varies from 33.33 to 31.44
per cent, the average for 236 eggs examined being 32.68 per cent.
_ “MARKET FRESH” EGGS.

The eggs from which the preceding information was obtained can
not be accepted as either a standard or an index of the eggs supplied
to the people for food, because modern conditions of living and
sources of food supplies make it impossible to furnish the market
with eggs of uniform quality and minimum age. It is necessary,
therefore, as already stated, to study the common market grades
of shell eggs accepted by the housewife and compare these with the
1 Loe. cit.
 COMMERCIAL EGGS IN THE CENTRAL WEST. 7

absolutely fresh eggs, as well as with the eggs put up for bakers 2.
use. Accordingly, ten open-market purchases were made of eggs as
they went to the consumer, and analytical data and candling records
obtained, using the same method as in the case of the fresh eggs.
The results are given in Table 3.
TABLE 3.—Eggs from grocery store.

Ammoni-
acal
Date of nitrogen | a: Se
Sample _ | Cost per ai 2 Size of | Description of sample un-
No. galls dozen. | Solcsor mathbd): sample. der candle.
we
basis
| |
1911. Per cent.| Eggs.
589 |Aug. 25.) $0.25 | ‘Strictly fresh eggs’’ car- 0.0011 24 | Fresh.
ton packed.
590: | 253d0i2 42. 320 |-Goodkeres/ 22 i.e kee. .0018 24 | Seconds.
pols esd 0). === B74 ee (eae (0LO een i Ps esse er aw . 0014 24 | Mixed firsts and seconds.
SO2ZF ees dO) 8s +205sies.2 :i a eS ECE RS Tang Sa . 0014 24 | 4shrunken eggs, 5 very stale
eggs, 1 blood ying.
636 | Sept. 5.. .25 | “Strictly fresh eggs’’ car- . 0010 24.
ton packed.
Oszi2e2dor ss P20GE |e SG OOd eges2e tee Fae . 0014 24 | 5 seconds.
OsSseead Once S20 ilzoeas COs ao ee eee . 0022 24 | 22 stale eggs.
BSOF |S dors as Re Se cee See Sa RSS ee Sh . 0021 12 | 9stale eggs, 1 hatch-spotegg,
1 blood ring.
640 |...do 20K 4) s“Hiresh eggs’ 72992
Pe os 0013 [esese ese 5 slightly heated.

19122 1/2 |
June 19 7 STS |DARD VLCCISS a Se a ae | . 0010 | 8 | Fresh.

Bacterially these eggs did not differ from the strictly fresh eggs.
According to the content of ammoniacal nitrogen they varied from
absolutely fresh to the usual stale, but not rotten, eggs. The price
usually, but not invariably, was in accord with the quality. The
carton-packed eggs were individually marked with the sign of the
producer, who had a reputation for quality to maintain.
RELATION BETWEEN BACTERIAL MULTIPLICATION AND CHEMICAL
CHANGES.

Experiments have shown that evidences of bacterial decomposition

can not be recognized by the sense of sight and smell until the organ-
isms have increased enormously in the food substance.
There is an interesting problem involved in the study of the rela-
tion between bacterial multiplication and chemical evidences of the
metabolic and catabolic changes that must accompany life processes.
Bacteria to the number of millions per gram have been considered
evidences of an altered chemical composition. Yet more recent obser-
vations would indicate that, for certain substances at least, the number
of organisms must approach the 100 million per gram mark before
the analytical methods for the detection of substances indicative of
bacterial life can be applied satisfactorily. Such, for example, has
been the finding of Hastings, Evans, and Hart,1 who have studied
1 Wisconsin Agricultural Experiment Station Research Bul. 25.
8 BULLETIN 51, U. S. DEPARTMENT OF AGRICULTURE.

the relation between bacterial multiplication and the formation of

lactic acid in milk; Burri and Kursteiner* state that the lactic-acid
organisms of milk may increase to 100 millions per cubic centimeter
before there is a definite rise in acidity. The same observation has
been made by Pennington? and associates when investigating the
decomposition of chicken flesh. In connection with this last study
it was found that a marked rise in the ammoniacal nitrogen content
e
E
of the flesh did not appear when the bacterial counts indicated a
few million organisms per gram, but at the next examination, when
the count was usually in the hundreds of millions, a rise was com-
monly found. The principles observed by the investigators cited
are corroborated by the results of the study of bacterial content
and chemical changes in eggs.
Not until chemical analyses show an increase in the ammoniacal
nitrogen are the senses able to detect infected eggs readily. Studies
discussed in detail in another section of this bulletin (p. 73) showed
that sour eggs which in the early stages are, with difficulty, detected
by the sense of smell, and eggs with light green albumen, which
are recognized by careful scrutiny, contain bacteria in many millions.
Eighty commercial samples which contained organisms capable of
producing gas from lactose in the presence of bile salt were examined
for B. coli, as described in the chapter on laboratory practice (p. 76).
A portion from each of the higher dilution fermentation tubes,
which showed gas, was plated on Endo’s medium, or on lactose litmus
agar, and from each plate having typical coli-like colanies several
were selected for examination.
Organisms conforming strictly to the definition for B. coli com-
munis, in the 1905 report of the American Public Health Association
on Standard Methods of Water Analysis, were isolated from 55 per
cent of the samples examined. Probably 15 per cent more contained
organisms which would be classed as typical B. coli by a majority of
observers because they differed very slightly from the definition just
given. In the remaining 30 per cent of samples, which were not found
to contain typical B. coli, the predominating gas-producing organism
was B. (Lactis) aerogenes. Of the typical B. coli organisms ex-
amined 81 per cent produced gas from sucrose. Practically all of
ihe organisms examined would be classed as members of the Colon
aerogenes group. If gas production from various sugars is sufficient
to distinguish between varieties of these organisms, a very large num-
ber of varieties were isolated. Some of them differed from any at
present described in the literature.
1Centralbl. Bakt., 1911, 2. abt., 30: 241.
2U. S. Dept. Agr., Bureau of Chemistry Cir. 70.

Webe
 COMMERCIAL EGGS IN THE CENTRAL WEST. 9

‘A detailed study of the organisms isolated is now in progress in

this laboratory. A speciai report will be issued dealing with this
phase of the work.
EXAMINATION OF EGGS OPENED ASEPTICALLY IN THE LABORATORY.

Because of the great diversity of conditions to which an egg in

the shell may be subjected, the corresponding variety in the results
which may follow, and because each individual egg must be considered
as an individual by the candler and the egg breaker, even though its
individuality is finally lost in the mixing, drying, or freezing of the
commercial product, the study to be reported here had to deal first
with single eggs of the various types found in commerce and which
may or may not be used by the breaker in his output intended for
food.
Tables 4 to 13 give the bacterial content of 300 individual eggs
and 26 small samples, aggregating 981 eggs, classified in accordance
with their most important or striking characteristic or the one prob-
ably responsible for the condition of the egg when it was examined.
For example, an egg might show a heavy, settled yolk in a sound,
clean shell, m which case it would be found in Table 5, under the
heading of “ Individual eggs with settled yolks.” But if that egg,
in addition to the settled yolk, had a dirty or cracked shell, it would
be classed in Table 7 or Table 8, devoted to dirty-shell eggs and
cracked-shell eggs, respectively.
The eggs were examined by means of a candle and their appear-
ance described before the contents of the shell were studied bacte-
riologically. The classification of the eggs was made on these ob-
served characteristics, and on others nceticed when the shell was
broken, rather than on the bacterial condition revealed by the labora-
tory work. The history of these eggs was known only in a very few
instances. None of them were “ market firsts,” or high-class eggs,
when they were received.
The study of the individual egg is logically followed by a study
of a number of eggs which are similar when graded by means of
the candle and by the characteristics observed on opening. Such
samples of hke eggs, aseptically opened, follow the report of the
individual eggs in a number of the tables. The technique used is
described on page 74. The samples were analyzed for the amount of
loosely bound nitrogen they contained, as well as for the number
of organisms.
STALE EGGS.

A very large proportion of the eggs going to the breakers are

simply stale; that is, the shell shows an enlarged air space, the yolk
has gained in opacity and definiteness of outline, and it is com-
10 BULLETIN 51, U. S. DEPARTMENT OF AGRICULTURE.

monly either above or below the normal meridian position. (See

Pl. II.) The white is frequently thin, and many times rough |
handling, combined with other age-accentuating conditions, have so
separated the membrane lining the shell from the membrane inclos-
ing the egg proper that the form and position of the air space can
change as the egg is turned. This appearance has caused the trade
term “ weak eggs ” or “watery eggs.” - But because these terms are
loosely used and have several meanings in different sections of the
country, or among different candlers, such conditions are charac-
ierized in this report as “movable air cells,” this term actually de-
scribing the change which has occurred.
In the late summer and autumn, when the lay has fallen off and
the country merchants are withholding stocks for coming high
prices, such eggs form a very large proportion of the current receipts
of the cities and are, of necessity, used by all of the community
who depend upon such sources for their ege supply.
When warm weather prevails many of these stale eggs show what
is termed “ heat”; that is, the yolk rises in the shell and is flattened,
the white becomes thinner than normal, and the air space increases
in size. These changes, well shown in Plate IT, take place in
fertile and infertile eggs, both of which become distinctly stale if
kept sufficiently long. In the fertile egg, however, there comes a
second series
f of changes, namely, incubation, which, in the very
earliest stages after the egg is laid, can not be distinguished by the
candling method, but has as one of its accompaniments the rising of
the yolk to the upper part of the shell. When the shell of a “heated”
egg is broken the germinal spot, if fertile, is seen to be slightly
thickened. In an infertile egg the germinal spot is seen with diffi-
culty. Such eggs, whether fertile or infertile, are good in appear-
ance, odor, and taste, but deterioration is rapid and they will not
stand long hauls to market nor keep well in cold storage. Hence,
such eggs are used in the egg-breaking plants. Their bacterial con-
tent is shown in Table 4, where it will be seen that the bacteria are
either very few or entirely absent, and that, bacterially, at least,
these eggs are not to be distinguished from eggs
oS
just laid.
 COMMERCIAL EGGS IN THE CENTRAL WEST. ik

TABLE 4.—Stale eggs.

INDIVIDUAL EGGS.

ite. Nkeon
nite tone Number of gas-
producing bac-
; teria per gram
4 : Date of Total number oppectoris pergramon | j, Terese pile:
Ne © | exami- P gan. Description.
ap nation.
Incu- Tncu- TIncu- Incu-
bated at |batedat} bated at | hatedat | White. | Yolk.
20° C. SiC. | e20eCk Bey,

1911
3023-1 | Mar. 28 0 0 0 Ol eh es ee As) ie Bae ee
3023—2)|easdose se 0 0 0 Qn ca ees ee Slightly dirty shell.
3023-4 |..-do.....- 5 0 5 CO) i te SO cl al re Clean shell.
3024-7 |Apr. 4 0 5 10 OUI A SSeS en aes Clean-shelled guinea egg
| kept in coid storage 1
year and in candling
room 1 week.
3026-1 | Apr. 7 10 Olea auuenes CO) Sea epee epee Clean shell, _ slightly
shrunken, small hatch
spot and floating yolk.
3026-7 |.-.do....- 20 0 10 CO) ROPESa OPS ie (ee ee Clean shell; slight shrink-
age. :
4005 | June 12 0 20 0 0 0 0 | Weak white and yolk,
hatch spot not en=
larged,movable air cell.
4039 ; July 8 0 0 0 0 0 0; Clean-shelled, fresh in<
fertile egg, kept at room
temperature for 8 days.
2 Weak white and yolk,
marked shrinkage,
slightly movable air
; cell.
4040 }...do..... 0 0 0 0) 0 0 | Clean-shelled, infertile
egg, kept at room tem _
perature for 8 days.
Watery white, marked
shrinkage, slightly
| movable air cell.

1 Samples 3001 to 3031, inclusive, were examined by Christine S. Avery.

SMALL COMPOSITE SAMPLES.

Total number
AesA iA Number Percentage of
of gas- | ammoniacal nitrogen
1M Ase 3 (Folin method)
ae Date of | Pp!ain agar. Brody: * | Per cent
ample |Source.!) exami- g of mois- | Description.
No. menor
‘ion. bacteria t ure.
Incu-. | Inen-, |Reh stam Ww
bated at/bated at in lactose et Dry
20°C. | 37°C. bile. basis. basis.

1912.
4579 | D 3} June 19 0 0 0 0. 0017 0. 0061 72, 13°26 3). .6) 2275) =
small, most
of them
shrunken.
4630 | E 41] June 28 50 200 0 . 0018 . 0064 72.02/66 eggs;
shrunken.
1911.
296/ B July 19 600 | 2,400 0 . 0016 ACG) eres
secsee So veneers.
/ shrunken.

1See p. 40.

Even when 5 dozen or more such eggs are mixed together the bac-
terial findings for the individual of the type hold good. The loosely
bound nitrogen in these samples of “stale” eggs is the same as, or a
little less than, that found in the “ grocery” eggs purchased in the
open market during the summer of 1911.
12 BULLETIN 51, U. S. DEPARTMENT OF AGRICULTURE.
EGGS HAVING SETTLED YOLKS.
Table 5 shows another type of deterioration, which is further ad-
vanced and more specific than that meant by the general term
“stale.” The yolk has become more opaque and consequently clearer
in outline when viewed by the aid of the candle and has fallen to the
pointed end of the shell (see Pl. 111), where it turns sluggishly when
rotated. Very frequently when these eggs are opened the yolk is
seen to have a very thin, weak membrane. During warm weather,
when incubation goes on almost continuously, though very slowly,
these eggs with settled yolks frequently show a germinal area about
one-fourth inch in diameter, having a visible white line through
their center—the “primitive streak” of the embryologists. (See
Pl. IJ.) Their odor is generally good and their taste not objection-
able, except for soft boiling or poaching.
TasLe 5.—Individual eegs with setiled yolks

White. Yolk. ee Number of

| @3- | gas-producing|
aS ERT EET SASS SS tee)
ota | Date of | Total number of bacteria per gram on gram in
“<= = | exami- | plain agar. lactose bile. Description.
i nation. FEE SSS
| Fncu- | Incu- | Ince- |} Incu- | Ineu-
| bated | bated | bat baied | bat White.) Yolk.
tat 20° C.'at
37° C.'at 20° C.lat
37° C_'at 37° C.
i |
1911. |
494 | Tune 12} 7 Oe ee Se ee Uy ee ee Watery whiie.
4913 | June 13 | 0 6 30 | 0: Ease: = 0 0 | Enlarged hatch spot;
i | | bloated yolk.
4914 |__.do..._-! 0 0 20 | Diba 0 0}
4015 |.-.do-_---- 30 0 20 10 0 @ | Enlarged hatch spot.
4016 |_..do-.-.- | 20 0 49 Ov <= 6 6; Enlarged hatch spot
i | bloated yolk.
4018 |.-.do..-.- | 0 0 30 | 51 a me 0 0
1912. | | | i:
ORG 7 Ane Stk eeea eS G2 (OR: ts eee Lae2 Marked shrinkage.
ECL
A gyArT ase fl Pee A | FV sil ee ng ee ee PALION enessrt
Res S Very weak yolk which
| broke when egg was
| | i | opened.
41089-12 |...d0.-..-- Pye. ero. Se eee eae oatLo) eee epee tie
ALOSSH1871 200-22 sce ee eee oe ee CAN). |== ose aes Marked shrinkage.
41089-19 |_..do_.._- e ee a te eee EEE SS (Pie 100) = 2. Marked shrinkace: yolk
bok | } i broke when egg was
- j | ' opened.
22 4 ie er 1 ee Aes Sees ees PEE eS eet Sai1 es | bee Marked shrinkage.
LORE 7s [SS eee ee ee ee ee fae ee OS0S00 Se Decidedly weak yolk
i | which broke when
[ | | | egg was opened.
ALIS8 j= do. oo ee eee WatsGhl coe 524 Co Ss Marked shrinkage.
ES ee ae oe ee ee eee Cabt0R oe ei! Do.
TCSSS i a FS Dp es (eee go Ee oe Ae age eel) | CN 25(1 | eee ell EP Do.
AURSS [220 ab 2 ee eee ee Os6300 bos S2-2 eeeepate Stes Marked shmnkage; yolk
| broke when egg was
: ' | } opened.
CALL LE 3Ol Breer; Fs eee egies Sune Pema eon [8 eee Cab IOQ = soa a eee shrinkage.
Al Oe a Cabin <<< Ee So
AAS Aas a. eee eee ft ee | Marked shrinkage; yolk Oat 400} =
}
broke when egg was
| | opened. ;
AACS45 ilo: does ea). Loo Bee 2 | ee ee eee Oat 100 2 ee Do.
AA OBSA46 ho ing ah oe eee ee ae ee eee a6 100 [= 2S eee Do.
41089-50 |_..do_-...- ees Perea ae, Career |e ais! 1) me | nee ee Marked shrinkage
ATR 5S ee ee eer ee eee ca MR Raa UE SL 1) eerie Beeietiorpece Do.
MOS 5s Sako? == 1 Sees so ies ee ee ee ee (O at 100 j-:._.-.- 1G 2s} Do.
HORT 57] do. Se ee ae Wat (hy ne ee Do.
41089-58 |...do-....- eee ges Be eeI oe oe eee. O36 IDR. 22k eines_ = ) Do.
UTS SR PR eS fee fal ee | SSE Ta Oa6 10 ee [eee see Do.
41089-61 |_..do....- pore Ar | wimps Ol gett ie eA ODE 100) 2-2 ea | Yolk broke when egg
| | Was opened.
ALOSS- GA ds oo ok ee eee 0 ab lOO fac2 0. =e | Marked shrinkage.
ANURS-63 4 dO. 2 ee ee alee O25 000 [222 -2=-- ae i aN
| |
10 at the dilution given indicates that no organisms were observed on the plate.
 COMMERCIAL EGGS IN THE CENTRAL WEST. 13

The bacterial content of these eggs is generally slightly higher

than in the earlier stage of staleness unless aging has occurred at the
low temperatures of the egg storage warehouse, when the count is as
low as or lower + than in new-laid eggs.
This type of deteriorated ege is very common in the breakers’
stocks, and, indeed, throughout the market seconds in both summer
and winter, due to rapid deterioration from high temperatures in the
one case and to the slow deterioration which occurs at 29° to 31° F.
in the other. The market life of such eggs is shorter than when dete-
rioration is not so far advanced. Hence the egg shipper with a
breaking establishment at hand prefers to break and freeze or dry
these eggs while they are still edible rather than to risk the certain
losses of a hau! to the consumer.
EGGS CHANGED BY INCUBATION.

Table 6 shows typical bacterial findings in eggs where deteriora-

tion had progressed along different lines than those described under
stale eggs and eggs with settled yolks. In the first group of 20 eggs
the development of the chick had not reached the blood-forming
stage, which normally occurs in about 24 hours when the temperature
is that of the hen, 103° F., but which had gone sufficiently far for the
candler to observe a small darkened area on the yolk. This in the
industry is known as a “ light spot,” and when the egg is broken it is
usually seen as a round area about three-eighths inch in diameter,
having two distinct zones, an inner and outer circle (see Pl. IT).
Such eggs, constantly sold in their shells for food purposes, are used
by the housewife without question if the odor is good and if the
white and yolk are intact. They do not ship well because the yolk
membrane is often weak, and many kinds of spoilage may develop in
them on short notice. Hence they are sent in large numbers to the
egg breakers in the producing regions.
1 Unpublished results on storage eggs.
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16
 COMMERCIAL EGGS IN THE CENTRAL WEST. LF

Bacterially, these eggs show a wider range and a slightly higher

average count than the two types previously discussed. All had the
yolk and the white intact. In 11 of the examinations reported the
white and yolk were studied separately. Two of the series were
sterile and not one showed the presence of B. coli. The fact that
the odor on opening these eggs was universally good is to be.
emphasized
The second series of Table 6 is composed of eggs which show before
the candle the presence of blood in the germinal disk, and which
are therefore equivalent to eggs at least 48 hours old at 103° F. In
all of the eggs in this series the embryo was either alive or very
recently dead, and the odor of the egz when opened was good. In
all of them the presence of blood could be distinguished by means
of the candle (see Pl. V). These eggs, in most instances, could
be separated into white and yolk. The average count for such eggs
showing blood rings, but without disintegration in the structure of
the egg itself, is low, and some eggs are sterile.
The third series, where incubation had continued for more than
the equivalent of 48 hours at 103° F. and where the embryo was dead
and the structure of the egg damaged to a greater or less extent,
shows a universally higher count than the other series and some indi-
vidual counts which are strikingly high. J&B. coli were noted but once
in this series. It will be observed that only one egg was separable
into white and yolk. The odor was sometimes good and sometimes

en EGGS HAVING DIRTY SHELLS.

The egg with a dirty shell is one of the most objectionable factors
of the egg industry. The contents may be fresh and the egg itself
may be large, but the dirt on the shell consigns it at once to the
seconds, and it brings a lowered price all through the market. Dirty-
shell eggs do not store well and are therefore not available for hold-
ing when the surplus production is greatest and when the market
can secure more good, clean eggs than it needs. Osten they scarcely
pay the expense of marketing. The breaker, therefore, removes the
dirty shell and endeavors to put the contents into a form in which
it can be marketed. The very objectionable filth on the exterior of
the shell naturally inclines one to the opinion that the contents of
the egg may also be contaminated. Such outer filth is not conclusive
evidence that the contents are infected.
Table 7 gives the bacterial findings for 51 dirty-shell eggs of
various grades of seconds and worse and 9 smali samples, aggregat-
ing 1,164 eggs. Winter, spring, and summer eggs are included. The
dirt on the shell consisted mostly of chicken feces and some dried
mud or dried egg. Some of the shells were stained. These stains,
which can not ordinarily be washed off, generally indicate that
water as well as filth has come in contact with the egg shell.
17625°—14—_2
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 COMMERCIAL EGGS IN THE CENTRAL WEST. 21

The contents of these dirty-shell eggs listed in Table 7 are varied.

Some show definite signs of incubation; some are aged eggs, as
shown by shrinkage; some show weak whites, some settled yolks,
etc. Bacterially this series, as might be expected, is very divergent
in the number of organisms observed. Two eggs out of the 51 con-
tain millions of organisms per gram in both white and yolk; 4 show
a count running into the thousands; 9 have more than a hundred
organisms per gram; and of the 86 remaining, 15, or 41.6 per cent,
are sterile. It is of great interest to observe that although so many
of these dirty shells were smeared with feces, which undoubtedly
contained B. coli, this organism was not once obtained in the body
of the egg, either in the white or in the yolk, and that it did not
appear in those eggs which had been wet is a still more striking
fact. This observation is in accordance with the work of Maurer.*
Sample 4083, with 3,100,000 organisms in the white, had a stale
odor on opening and a stained shell, which looked as though it had
been wet.
Sample 4128 had a very badly brown-stained shell, and though
the odor of, the egg was good on opening, 72,000 organisms per
gram were found in the white. Sample 4137 showed 4,600 organ-
isms per gram in the white, which may, perhaps, be accounted for
by the stained shell. On the whole it is the stained shell which
seems most likely to be the offender, an opinion which is strength-
ened by the observation that where there is a count of any magni-
tude it is not only most commonly in the eggs having stained shells,
but the organisms are usually more abundant in the white of the
ego than in the yolk, from which one might infer that the infection
is from the exterior.
The nine samples, composed of more than one dirty-shell egg, vary
in bacterial content just as do the individual eggs. The chemical
analysis shows that the loosely bound nitrogen varies with the char-
acter of the egg, not with the quantity of dirt on the shells. Sample
4631, for instance, shows 0.0024 per cent of nitrogen and is described
as very stale, but it was not any dirtier than Sample 553, wherein
only 0.0013 per cent of nitrogen was obtained.
Tf one may draw any conclusion from the findings set forth in
Table 7 it must be that the dirty shell; per se, is not a sufficient
ground on which to condemn an egg, though the odor of the egg
when opened should be carefully observed, especially if the shell
shows stains or other evidences of having been wet.
1 Bacteriological Studies on Eggs. Kansas State Agricultural College Bul, 180, 1911.
22 BULLETIN 51, U. S. DEPARTMENT OF AGRICULTURE.
EGGS HAVING CRACKED SHELLS.

Mechanical injury to eggs, due to rough handling, is another great

money loss to the egg industry and a food loss to the consumers.
Many eggs are completely wrecked and are termed “ mashed” eggs
by the industry. In this case they are not only lost but they soil a
number of eggs otherwise good. Where the shell is so broken that
the contents are escaping the egg is termed a “leaker.” In addition |
to leakers the industry has to contend with enormous numbers of
“ checks "—that is, eggs which have cracked shells but intact mem-
branes. _
Leakers are thrown out at every stage of handling, from the coun-
try merchant or egg peddler to the city retailer, and are generally a
total loss. Checks are disposed of if possible and as near the gather-
ing point as may be, because they are weakened mechanically and are
free food for any bacterium or mold that may chance to fall into the
erack. Hence they are sure, In commerce, to rot quickly. The
cracked eggs coming to the breaker in the producing section are re-
ceived by him much sooner after the damage is done than by the
breaker in the consuming center, and for that reason they are usually
in better bacterial condition. Table 8 gives the results of the exami-
nation of 56 of these cracked eggs, both blind and visible checks.
Some of these eggs were fresh and above reproach except for the
damaged shell; others had dirty as well as cracked shells, hatch
spots, weak yolks, thin whites, etc.
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 COMMERCIAL EGGS IN THE CENTRAL WEST. DAT

Like the eggs with dirty shells, there is a wide variation in the
number of bacteria, though only 5 out of the 56 individual eggs listed
are sterile in a dilution of 1 to 10, and when the count is beyond a
few hundred organisms per gram it is generally very high—that is,
in the hundreds of thousands or millions. Four eggs have a bacte-
rial content which is much higher than that observed in the other
52 eggs, ranging from 2,700 to 370,000,000 per gram. In three. cases
out of the four an objectionable odor was noticed when the egg was
opened, and three of these eggs had dirty as well as cracked shells.
B. coli, though sought in all but five of the samples, were found in
but three instances and then in eggs which showed a high count.
Here, again, the whites of the heavily infected eggs show a much
higher count than the corresponding yolks. This is quite in line with
the origin of the infection.
The samples from small lots of eggs with cracked shells bear out
the findings from the individual eggs. Where the eggs with cracked
shells are of good quality, both chemical and bacterial analyses indi-
cate that fact. Where deterioration has begun, the cracking of the
shell does not materially alter its course, but it hastens decay. Of
course, the protection which the shell affords is lessened by cracking,
and bacterial invasion is only a question of time and environment.
EGGS HAVING THE YOLK SEEPING INTO THE WHITE.
During warm weather, when the deterioration of eggs proceeds
with great rapidity and in the most diversified fashion, many eggs
are received at the concentrating centers, especially those reached
by railroad or where the wagon haul is over rough roads, which show
on candling filaments of yolk that have apparently found their way
through apertures in the vitelline membrane for longer or shorter
distances into the white.
Sometimes these filaments are very few and distinct, half an inch
or more in length; in that case the egg white is usually normal in
color, even between the filaments. Sometimes the seepage of the
yolk into the white might be better described as diffuse, in which case
very numerous and tiny filaments make.a yellow zone around the yolk
membrane, the outer portions of white remaining clear and the usual
color. As the process of mixing progresses the white becomes more
and more yellow and the vitelline membrane less and less resistant,
until finally the latter ruptures and a complete mixing of yolk and
white follows.
Even the most careful cracking of the shell at its equator is at
times sufficient to rupture the yolk membrane extensively, thus per-
mitting the yolk to escape entirely. At other times a fairly clean
_ separation of white and yolk can be made. Generally such eggs ex-
hibit, in addition to the filamentous yolk, distinct signs of age, such
as shrinkage, and of rough handling, as shown by the movable air
- cell. Ordinarily the odor is good or somewhat stale, the sort of odor
28 BULLETIN 51, U. §. DEPARTMENT OF AGRICULTURE.

that the housewife terms “ eggy ” or “strong.” If the odor is not

bad and if the mixing of the yolk and white is slight, the housewife
uses these eggs for general cookery, though when the whites are to
be whipped separately to lightness they are not satisfactory. The
egg shipper does not attempt to send such eggs on long or hard jour-
neys, because this mixing is accelerated by jarring; nor does he at-
tempt to store them, because white and yolk continue to mix more
and more rapidly. These eggs are therefore used very largely by
the breakers. They should be examined very carefully for odor and
appearance when broken, as they may be incipient forms of the “sour
egg” (see p. 61). Table 9 illustrates the condition of this series.
TABLE 9.—EKggs with the yolk seeping into the white.
INDIVIDUAL EGGS. 7

Total number of bac- | Number

teria per gram on plain of gas-
commie Date of | agar incubated at— producing huey
N he exami- bacteria Description.
i nation. per aie
in lactos
20° © 37°C iaig

1910.
3006-4 | Dec. 12 77, 000 8350009] bays
sai a
3006-5 |..-do.. 39 LOOK beanies
3010-2 |...do.. 25 Daye Ss oe Peon Sas

1911. |
4003 June 12 10 10 0 |
111-2 | June 21 5 15 0 | Marked shrinkage—fixed air cell.
HES} excl sae 60 0 0
HAs domes: 5 0 0
111-51. -.do. =. 30 0 0
111-6 |..-.do...- 10 0 0
At fe eadoness 0 0 0
122 | June 24 10 0 0 | Early stage.
12AS ee dOnese 5 5 0 Do.
125) adore 5 0 0 | “Strong’’ odor.

1912
136 | June 27 50 | 30 0 | Early stage.

1911.
Pale |ra
fuubys. Ital 10 10 0
4103 | Nov. 1 5 90 0 | Clean shell; stale odor; marked shrinkage;
movable air cell.
4108 | Nov. 7 2,000,000 | 1,700,000 Shell clean, but not fresh looking; stale odor;
marked shrinkage; movable air cell.

SMALL SAMPLES.

po ;
number o aBysen
bacteria Number pee s . f 7
gram : ize of sample.
On alain of gas- | Gelatin nitrogen, Per ea eg
Sample Date
aneAOof agarrincn.
incu |producingt |liquefying} Folin method. | cent | Ether
No exami- | hated at— | bacteria | organ- of ex-
: nation. per gram | isms per mois- | tract.
in lactose} gram. ture,

es Wet | Dry Bacte- |Chem-

20° C2137" © basis. | basis. Tiolog-
ical.
| ical,

1911. Eggs. | Eggs.

115 | June 23 3 3 0 OP aa and eee ese 72.35 | 10.96 12 24
170 | July 3 100 5 0 0 | 0.0033 | 0.0111 70.31 12. 45 12 24
Ia s2dor 90 0 0 Onl eO033 ile Oll25 |eZ OuG Onell 1 4
172 |...do- 100 RRR AA A Sead re case oos .0032 | .0114] 71.84] 10.86 1 4
Lisne=- do: 100 35 0 0} .0029 0103 | 71.78 | 10.55 1 £
174 |...do 180 5 0 0] .0038) .0141 | 73.12] 10.01 1 4
 COMMERCIAL EGGS IN THE CENTRAL WEST. 29

Two of the eggs show decidedly high counts. The first, Sample
3006-4, was a winter egg and had probably been held by the farmer
or merchant for a long time. The second high count, Sample 4108,
was a July egg, and its bacterial condition might be explained by
the fact that the shell showed signs of much handling, and the egg
had acquired a stale odor. The other samples of the series show low
counts; there was an absence of #. coli throughout, even in the case
of the two high-count eggs.
Six small lots of eggs, where deterioration had gone further than
in the type just described, were also examined for bacteria and loosely
bound nitrogen. The number of organisms was negligible; the
amount of loosely combined nitrogen was higher than had been pre-
viously noted. All of these eggs would have been discarded by a
careful grader because of the yellow color of the white. Just where
to draw the line, however, is not a simple matter. Practical expe-
rience would indicate that when the white of the egg was normal in
color and when the filaments of yolk were entirely distinct from the
white, or when, if the seepage was by the diffuse rather than the
localized method, the outer zone of egg white was normal, the bac-
terial content was low and the loosely bound nitrogen did not rise
above 0.0038 per cent.
WHITE ROTS.

If the egg, where white and yolk are just beginning to mix by
either method of seepage, be held under commercial conditions, it
becomes what is known to egg candlers as a “* white rot,” or to some
as a “sour rot,” but the latter is a misleading term and should be
discarded. The inexpert or careless candler fails to notice these white
rots; hence they are too often found in the breaking room; when
opened yolk and white are seen to be completely, or almost com-
pletely, mixed. Very frequently the mixture is much thinner than
the mixed yolk and white of a fresh egg and may or may not have
an offensive odor. Its appearance is never appetizing. Sometimes
scraps of membrane are seen, suggesting a disintegrated embryo;
again, the contents are thin, homogeneous, and pale yellow (see
Pl. VII). The series of eggs given in Table 10 is typical of eggs
having these characteristics.
30 BULLETIN 51, U. 8S. DEPARTMENT OF AGRICULTURE.
TABLE 10.— White rots (individual eggs).

Total number of bacteria |Number

per gram on plain agar | of gas-
incubated at— produc-
Sample
N= pu Aer es ing bac-
teria per a9
Description of sample.
gram in
ACs 37° €: lactose
bile.

3006-2 | Dec. 12,1910 | 110,000,000 15,000,000 |---------- Pale yellow.contents.

S006=3 [eee oe GOte athe ee 80,000, 000 |----------
3012-4 |Jan. 14,1911 Ole: OBE |sce secee
275 | July 3,1911 5, 500, 000 3, 300, 000
495 |Aug. 10,1911 (4) (1) 10, 000+
ASG aes: dose ee 0 0 0
4O7 ieee: dostaas (1) (1) 10, 000--
498 |... doses: : 120, 000 120, 000 0
ys ees ee SS COE sss 0 0 0
5OOU een Goes.(0) (1) (1) 10, 000+
4070 | Sept. 5,1911 | 180,000,000 | 270,000,000 100+- | Clean shell; marked shrinkage: mov-
able air cell.
4110 |Nov. 1,1911 160 180 0 | Shell slightly dirty but fresh looking;
stale odor; marked shrinkage; mova-
ble air cell.

1 Innumerable; dilution, 1: 10,000.

The bacterial content in 8 out of the 12 is very high. Two of the

egos were sterile and two of them showed a low count. B. coli
were looked for 9 times out of the 12 and found 4 times.
The high bacterial content of these white rots is quite in accord
with their appearance. Why there should occasionally be a white
rot with a low count, as in Samples 3012-4, 498, and 4110, or even
a sterile white rot,) as in Samples 496 and 499, remains to be ex-
plained. Since these white rots seem to be the logical sequence of
the mixed egg, they might easily parallel the latter in their bacterial
content,
EGGS HAVING YOLK ADHERENT TO SHELL (SPOT ROTS).—
The “spot rots” of commerce are eggs in which the yolk has
become adherent to one or both of the shell membranes and, per-
haps, to the shell itself by means of the membranes. When held
before the candle, therefore, the yolk is seen as a distorted, deeply
colored mass pressed against some part of the shell (see Pl. TV).
As the egg ages in temperatures which are lower than those causing
incubation phenomena, the yolk of either the fertile or the infertile
egg settles. If the egg is not moved the yolk finally adheres to the
membrane against which it rests and it becomes a “ spot rot” or, as
termed in this report, an egg with the yolk adherent to the shell.
If the egg is infertile and ages at such temperatures as prevail in
summer time, the yolk frequently rises, presses against the air cell,
and finally sticks there. Forty-two such eggs are listed in Table 11.
When held before the candle some show no marked characteristics
except the adherent yolk. Others show distinct evidences of incuba-
tion, general deterioration, cracked shells, ete.
1In a dilution of 1 to 10.
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Nine of the 42 eggs—or 21 per cent—show very high counts, the

maximum being 320,000,000 bacteria per gram in the white of Sam-
ple 4049. The lowest count in these infected eggs is 150,000 per
gram in the white and 94,000 in the yolk. Then there is a sudden
drop to 1,800 per gram in Sample 3014-5, and the 18 remaining
samples—or 43 per cent—which show bacteria present have so few
that they may be neglected for practical purposes. Fourteen sam-
ples—or 383 per cent—were sterile in both yolk and white. The
organisms in Sample 3012-1 were probably from the dirt spot on the
shell against which the yolk had lain. Sample 3017-1 had a sour
odor which indicates bacterial contamination in quantity ;3017-2 had
a dirty shell; 4030, 4049, and 4157 had objectionable odors on .open-
ing; 4141 had a stained shell. Samples 3017-2 and 4021 had no
distinctive feature except the adherent yolk.
MOLDY EGGS.

Damp cellars, wet nests, stolen nests, etc., are responsible for the
condition of eggs which show, on candling, dense black areas of vary-
ing sizes inside the shell. When the eggs are opened these areas are
found to be infected with a mold, usually a common green mold, of
the Penicillium family (see Pl. VI). Such eggs almost invariably
have a moldy odor. If the mold spot is small it may not affect the
integrity of the egg structure; on the other hand, it may grow to
such dimensions that no distinction between yolk and white can be
seen.
The products of the growth of the mold may gelatinize the white
or liquefy it and may coagulate the yolk into a cheesy mass or render
it watery. It is, however, but seldom that a mold in pure culture is
found inside an egg; generally bacteria are also present, and some-
times in large numbers. Both the white and the yolk of moldy eggs
are apt to be discolored, usually becoming brownish. This color is
not always confined to the area of visible mold, but may be diifused
throughout, as shown by cultures made from white and yolk remote
from the visible infection. These and other characteristics are noted
in Table 12, where the results of the examination of 45 individual
egos showing mold visible to the eye are recorded.
17625° —_14——_3
 BULLETIN 51, U. S. DEPARTMENT OF AGRICULTURE.

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37
88 BULLETIN 51, U. S. DEPARTMENT OF AGRICULTURE.

It is interesting to observe that B. coli were found in but 2 of

the 45 eggs; that in those cases it was present in small numbers
only and then with large numbers of other bacteria. In fact, the
mold in Samples 4029 and 4032, respectively, was in the air cell and
apparently had not penetrated the egg membrane, though undoubt-
edly it would have done so in time. It is also of interest to note that
the odor of Sample 4029 was putrid and that of 4032 stale; both had
volks stuck to the shell.
Molds, as mdicated by the descriptions given in Table 12, may
appear in clean and dirty shell eggs. If the shell is dirty, the first
visible spot of mold is very often beneath the spot of dirt. Since the
mold infections seem to be due almost entirely to shell penetration
after laying, one would expect to find the egg—both yolk and white—
adhering to the membrane. Such is usually the case if the growth
is extensive.
So varied are the visible results upon the egg of the growth of
mold inside the shell that much space might be cna describing
individual eggs. The salient points for our purpose, however, are —
the facts that the eggs which show mold before the candle give a
growth of mold when the egg substance is transferred to suitable
culture media, anda study of the substance of such eggs shows that
the mold is not confined to the area where it is visible, but is com-
monly diffused throughout both white and yolk. A moldy egg is
also likely to show a large number of bacteria present.
BLACK ROTS.
*
é
Black rots need but short comment here. They are recorded for ay
|
comparative purposes only. Table 15 gives the bacterial findings in :
10 of them. The odor and appearance, both before the candle and
after opening, would exclude their use for any food purpose or even
AE
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for leather tanning. They could be used fer fertilizer.
‘ABLE 15—Black rots—individual eggs.
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Dateate ofol | Sens se ai—

azar incubated 0 pian aes
Baraple |examina- iced Description.
INO: > ta~icn.
2? GC of

| 1910.
3007-6] Dec. 21 | 180,000,000) 140, 000.000/---.---..--- | Thin, wWaiery contents, with bad odor.
3009-5| Dec. 27 49, 000, 20 7.000: C0 | Watery contents, with strong edor.
...do....- 4, 200, 000, 000/6, 300, OCD, 000,3
3009-6! eee SS | Watery, olive-green, gassy contents, with bad
odor.

3010-1; Dec. 28 | 120, C00, 000 33,000, C00 “i aes | Green contents, with strong odor.
| }
|

191i. |
3012-3) dan. J4ie see 790, 000, 000) - -----------] | Brownish, gassy contents, with bad odor.
4045) July i1 | 350,050,600 340, 000, 00C) 1,000,000+)| Black under candle; egg had a bad odor before
| | being opened, and a still worse one aiter-
} wards; some shrinkage; fixed air cell.
 COMMERCIAL EGGS IN THE CENTRAL WEST. 39

TABLE 153.—Black rots—individual eggs—Continued.

| Totalnumber of a |
| Number of |
ria per gram on plain | gas-pro- |
Date of | agar incubated at— : :
Sample)
Ree jamin
ee | = |dems bac |
teria per Description.
| | j gram in lac-)
ee 207E. 4 3r° C. | tose bile. |
j
wee re
4059) ‘July i3 | Gf 000: (NNN | 1,000,000 Black under eandle; contents were very Zassy
| | and had airightfulodor; clean shell: marked
t shrinkage; movable air cell.
4066,_._do_-..- 11, 000, 000 5,400,006 1,000,000+| Black under candle; bad odor on opening;
| | clean shell; marked shrinkage; movable air
j | eek
4087 Oct. 9) 660,000,000 180,000,000 0 | Black under candle; shell dirty and stamed in
| | | one spot with a damp feather, underneath
| which was a mold spot; balance of the egg
‘ Was 2 Dlack rot.
4114; Nov. 7} 160,000,000 280,000,000 100 BElack under candle: strong odor of hydrogen
| sulphid: mside of shell and shell membrane _
| black: shell not fresh looking; one-third
} | | shrinkage.

The maximum bacterial count was 6,300,000,000 per gram; the

minimum 5.400.000. #&.coli were looked for five times and found feur
times—in very large numbers except in one sample.
COMPOSITE SAMPLES OF EGGS OPENED COMMERCIALLY
IN TRE PACKING HOUSE.
The study cf eggs opened aseptically in the laboratory is eee
tcllowed by a study of eggs broken commercially in the packi
house. For this ipower ere a large number of samples were taken
of the various types of eggs pope throughout the egg-break-
ing season of 1912. It was hoped that the laboratory results, to-
-gether with the characteristic appearance and odor of the diferent
_ classes of eggs, would givea practical working basis for the grading
| .of eggs used in the preparaticn of frozen a desiccated eggs.
| The commercial Panton: under which the eggs were broken
| are described in the discussion of D, E, and F houses for 1912,
' in a forthcoming Department of Agriculture bulletin. The method
of cpening was in brief as follows: The eggs were broken on a ster-
ilized knife edge, the two sections of the snell pulled apart with the
_ thumb and first and second fingers of each hand, and the contents of
' the ege allowed to drop into a sterile cup. After every infected egg
| _ which could be detected by the senses, the eperator poe the knife
P- and cup with sterile ednpmennt and eel and dried her hands.
| ‘The fingers were kept dry by means of tissue paper or small towels,
which were used but once before launder ring. By this method the
contamination of the liquid egg during the process of breaking was
reduced to a minimum.
| If the sampie consisted of less than 9 eggs, the liquids were poured
| directly from the cups to a sierile 16-ounce salt-mouthed bottle,
40 BULLETIN 51, U. S. DEPARTMENT OF AGRICULTURE.

mixed by shaking, and a small portion of the mixture was trans-

ferred to a sterile 4-ounce bottie contaming pieces of sterile glass;
the large sample was for chemical analyses, the smaller one for
bacteriological examination. If the specimen represented 9 or more
eggs, the eggs were collected in a suitable container and mixed, a
bacteriological and a chemical sample being taken. The samples
were frozen in a sharp freezer 12 hours or less, packed im chilled,
cork-insulated boxes especially constructed for the purpose, and
shipped by express to the laboratory, where they arrived hard
frozen. About 3 hours were required for transportation from E
house and about 12 hours from D and F houses. |
The samples were taken on the successive weekly visits made at
D, E, and F houses during the season of 1912. <A section in each of
the tables containing the laboratery results of this research indicates
the time and place of sampling. For example, F—5, in Table 14,
signifies that the sample was taken on the fifth visit to F house.
The number of eggs represented in each sample varied from 4 to
360.
JULY AND AUGUST FIRSTS.
Since strictly fresh eggs are not used in the United States for the
preparation of frozen and desiccated eggs, no studies were made of
this grade opened under commercial conditions. During the latter
part of July and August, however, when receipts are not only light
but also low in quality and cheap, eggs commercially graded as firsts
are sometimes used to piece out the regular supply of breaking stock.
The firsts of the summer and autumn months differ before the candle
from the firsts of the spring months in that the former are much
more shrunken, the yolks more opaque, and the whites are less firm.
Firsts, also commercially termed “storage-packed No. 1 eggs,” con- 7@
stituted a large percentage of the breaking stock used in F house
during the season of 1912. They were graded from the daily re-
ceipts and held in chilled rooms until needed to fill out the regular
quota of eggs for the breaking room.
On the last two visits to F house samples were taken of the liquid
egg broken from five lots of firsts, each representing 15 dozen eggs
(see Table 14). Care was taken during the process of breaking to
eliminate all eggs which might have a deleterious effect on the liquid
product. For example, from one sample an egg with a green white
was discarded and from a second an egg with a broken yolk which
at one time had been adherent to the shell. Some of the firsts had 7
cloudy whites, but such eggs were not discarded in the preparation 7]
of these samples because laboratory studies proved that they were "|
not infected.
 COMMERCIAL EGGS IN THE CENTAAL WEST. Al

The laboratory data given in Table 14 show that three of five

samples contained less than 1,000 bacteria per gram at 20° C. and
the other two 25,000 and 92,000, respectively. In three samples B.
coli were not found and in the remaining two they were present in
small numbers. Since the bacteriological findings given in Table 4
indicate that this grade of eggs, when opened aseptically, 1s prac-
tically sterile and contains no B. coli, it might be concluded that
the organisms found in the samples opened in the packing house
were referable, for the most part, to outside contamination and not
to the eggs themselves. On the basis of this assumption these re-
sults will be taken as a standard of comparison in the succeeding
discussion of the bacterial contents of other types of eggs.
The moisture content of four of the five samples of firsts was lower
than the amount found in fresh eggs, which result would be expected
from the difference in the amount of shrinkage in the two types of
eggs. The average percentage of ammoniacal nitrogen, which was
taken as the index of protein decomposition, was 0.0020 per cent on
the wet basis for summer firsts, compared with 0.0013 per cent for
absolutely fresh eggs. These figures show well the difference in
quality of the two grades of eggs.
TaBLe 14.—July and August firsts.

[15-dozen fots.]

Percentage of am-
Total number of bacteria per gram on | Number | moniacal nitro-
plain agar incubated at— of gas-pro- gen, Folin
Date of ducing method. Percent-
No. | Source.} collec- bacteria age of
tion. Be es moisture.
-in lactose
° ° . Wet Dry
PO OO} BCs bile. inasicl aces

1912.
4802 F 5} July 22 Less than 1,090 Less than 1,000 10 0.0022 | 0.0072 69.59
48°3 F5 | July 23 25, 000 20,060 0 -0020 | .0065 69.14
4966 F 6 | Aug. 13 Less than 1,000 Less than 1, 000 0 -0020 | .0074 72.84
4967 F 6 |...do..... Less than 1,000 Less than 1, 000 MICs) ME aa Sy ga a
4985 F 6 | Aug. 15 92, 000 1,000 0 .0019 | .0068 71.87

SECONDS.

Seconds constitute a large proportion of the eggs used in the

frozen and desiccated egg industry. In the spring, before the
candling season begins, this grade consists of small, dirty, and over-
sized eggs sorted from receipts by inspection. After the first of
June, when all incoming eggs are graded according to the condition
of the contents, seconds also include shrunken eggs, hatch spots,
weak eggs, heavy rollers, etc.
Since dirty shells during the process of opening contribute a
special source of contamination. to the liquid eggs, this class of
egos is considered separately in this dissertation.
AQ BULLETIN 51,/U. S. DEPARTMENT OF AGRICULTURE.

_ During the interval between May 4 and August 30, 1912, 9 sam-
ples of whites, 9 of yolks, and 25 of whole eggs were taken from
the product obtained from seconds, the different lots varying in size
from 6 to 30 dozen eggs. During the process of breaking care was
taken to discard all eggs whieh, from appearance or odor, were
abnormal. The first five samples given in Table 15 under the sec-
tion devoted to whole eggs, and the first four in the part assigned
to whites and yolks, represent the product broken from small and
oversized eggs. The maximum count of organisms per gram was
34,000 in the whole eggs and 85,000 in the whites and yolks.
The counts of the product from small and oversized eggs were
no higher than those found in the hquid eggs broken from July and
August firsts; in fact, the amount of ammoniacal nitrogen in the for-
mer was lower. The chemical results show, therefore, that the con-
tents of the spring seconds were fresher than the summer firsts.
The samples taken after June 1, 1912, represent the small and
oversized eggs sorted from receipts by inspection and also those
showing incipient deterioration as observed by the candle. An
observation of the laboratory results in Table 15 shows a general
trend toward higher bacterial counts and greater amounts of am-
moniacal nitregen as the season advances.
TABLE 15.—Seconds.

WHOLE EGGS.

| |
| Total number of | __ Percentage
| | bacteria per |Number) of ammoni-
| | gram on plain ieaoe acal a Dae
Sam- | Date of agar incubated | Produc-| gen, Folin | cent- :
ple |Source.) collec- at— a pac method. | age of Sis a Remarks.
No tion ees mois- mple.
Pram |e |e aee
‘ : lactose Wet |Dry 3
20° C. 37° C. bile. | hasis.| basis.

1912. :
4245 Fi1| May 4 300 750 WO esase |Bei bape ereraene 15dozen...
A249 D1 Mery eetOR |ereese 1, 100 10,0. 0016,0. 6948) 66.23]...do..
A259 | Di1| May 7 400 150} TOUS ssrea ey ech | Cet |e Ones
aba Dole dor 34, 060 400 Si AR ame dose ae
A411} D2| May 27 23, 500 17 O00 ieee ON OOO Peapaeres |r eee 10 pounds.
A578 D3 | June i9 600 300 0) .0016) .0959) 73.00} 6 dozen...
A633 F 4, June 28 2, 500 950 0! .0020} .0068! 70. 45)...do...-..-
A694 D4 | July -9 27, 500 21, G60 1,000) .0021) .0079} 73.35} 12 dozen...
A697 DAM PR Oteene 5, 200 300 0) S002 ROOMS | (en Sill See eee nee
4708 D4 | July 10 10,500 9, 900 1,000) .0019} .0067| 71.84) 12 dozen...
4710 D7AR | Osere: 3, 200 2, 100 0} .0015] .0055} 72. 75) 15 dozen...
4803 F5 | July 22 110, 000 93, 000 100} .0023) .0083] 72.38 GOs
4833 F5 | July 24 1 4,000 1,000} .0018} .0055} 69.39 MO sens
4945 E6 | Aug. 9 850, 000 7, 000 . 0023] .0084] 72. 63} 6dozen..
4957 F 6] Aug. 12 66, 000 38, 500 10} .0015} .0050} 70.16 Ses g
A972 F 6 | Aug. 14 750,000) 550,000 0} .0017| . 0061) 72.30)...do..----
A1002 D 6 | Aug. 19 2,000) 0 in 1, 000} 0} .0020) .0072) 72.31) 24dozen...
41018 D 6 |Aug. 20 600, 000 550, 000 100} .0018} .0057| 68. 25 2 pounes.
41019 Oe cOlOs aoe 39, 500 ZAS O00 |Se 22 Beat ee Ee re eS eae ee oe Onu seer
41029 D6 | Aug. 21 320, 000 270, (O00) Pen) = SESS 5c aclsosecd|\sac GoOccesee
A1031 1D) OABGAC As 5 6 500,000) 500, 000 100} .0017| . 0061 72.53) 24 dozen...

ti070| EY |Aus. 97 |3.600,000)

41086 | E7]| Aug. 30]
1,600,000
'430,000| | 55,000)
WL || gage dono,
100) . 0026} .6095| 72. ae doxsenus

1 Less than 1,000.

 COMMERCIAL EGGS IN THE CENTRAL WEST. 43

TABLE 15.—Seconds—Continued.
WHITES AND YOLKS.

i ] j |
Total number oi |__ eras |
| | bacteria per |Number of ammoni-|
| gram on plain | °fgas- | acal nitro- |Per- |
Sam- Date of agar incubated |Produc-| gen, Folin Es Rana ot
ple
No.
Source.) collec-
tion
|| at— ing bac-|| “method.
teria per
jageof|
mois-|
Ove?)
pie.
Remarks.
| 5 |Spee Jeena Rls
Wet ||Deel
Dry pos
/ 20 Cc: 37 |On { bile. |basis.
oe j

|
a rs | a
| 1912 | |
4363 | F2|May 2] 49,000 1, 200) Oisc: Feleae oe i 15dozen.... Whites.
ASG | ee) ee Ease.e| S, O66} 1,60 Or aE eee 1. epee eee Yolks of 4383.
4493 | D2| May 27 | 1, 600) 150 0.0. 0004/0. 0031) 87.10 15 pounds.) Whites.
4494} D2|._.do.....]. 1,200 400 10} 0033] .0072| 54.15]... ---. Yolks of 4403.
4416 | D2}_.-Go.....| 60,000] - 20,000; 100)... Laie. ee | 25pounds.) Whites.
AAT D2 | do... 15, 000) TL ace
ea ena [eens lest bee as ile Yolks of 4416.
4423 D2) May 28 1, 060) 1, 060) id) . 0903) .0023) 87.14) 10 pounds.| Whites.
4424 1B) PEGI es 4, 000! 2, 700) 1, 000) 0030) aay SY(oc5)|Rees
See eae Yolks of 4423.
4868 | D5 | July 30| 360,000} 330,000, 1, 000).-..-. [epee Ee welts | 13 pounds. |Whites.
ABEGa =D 50 |=. do... - 110,000, 90,000; @) |. 37| - 0083) 55.42) 11 pounds.| Yolks of 4868.
AQT7T F6 | Aug. 14 40,000} 40,600 100 ok assee Reese | 20 pounds.| Whites.
4978 F 6 |...do....-; 130,000; 130,000 0 - 0037) O08 7) aa. 25|-- dor...o- | Yolks of 4977.
41005 | D6| Aug. 19| _ 27,0001 22,000) 1, 000)....-- ee eae ae eee | Whites.
41006 DGhte Ga.e ce 300,060} 180, 600 1,000; .0029) .0067 56.66) 15 pouncs_| Yolks of 41005.
47011 D6 |_..do....-| 608,000) 650,600 0} .0003) .0523' 86.96) 18 pounds. | Whites.
41012 D6 |_-.Go_.-:.| 650,060} 600,000 C) .0030) .G065 53.64) 13 pounds.) Yolks of 41011.

2 i Less than 100.

The lowest number of bacteria found at 20° C. was 300 per gram
in a sample taken during the early part of May, and the highest,
3,600,000, in a specimen obtained during the latter part of August.
Of the 16 samples of whites and yolks none contained over 650,000
bacteria per gram; of the 24 lots of whole egg only about 8 per cent
had more than 850,000. There were no B&. colz in 14, or 35 per cent,
and 100,000, or less, in the remaining samples. The percentage of
ammoniacal nitrogen varied from 0.0015 per cent on the wet basis
in spring seconds to 0.0026 per cent in summer seconds.
A larger number of eggs was discarded while breaking the summer
and fali seconds than when opening analogous eggs in the spring.
It is probable that the organisms in the samples of liquid egg from
seconds are referable partly to outside sources, but chiefly to the eggs
themselves.
EGGS HAVING DIRTY SHELLS.
The highest percentage of dirty-shell eggs occurs during the wet
spring weather. Since they do not keep well in storage, a large num-
ber find their way to egg-breaking establishments.
| Table 16 gives the laboratory findings of six samples taken during
EF May, June, and August, 1912. Fifteen dozen eggs were represented
in one specimen tid 6 dozen in the other five. All of the egos were
-eandled before opening. The number of eggs discarded during the
process of breaking gives, perhaps, an index to the quality of the
eges in the ne coos. Not more than three decomposed eggs
were eliminated from the samples taken during May and June and
not more than six from the two specimens broken in August.
44 BULLETIN 51, U. S. DEPARTMENT OF AGRICULTURE.
TABLE 16.—Eggs having dirty shells.

[6 to 15 dozen lots.]

| | Total number of bacte- |Number | Percentage of am-

| | ria per gramon plain | of gas- moniacal nitro- =
|Date of | ae cubated ee producing} gen, Folin meth- | per cent Size of
No. |Source.) collec- | bacteria od. of mois- ae 4
| tion. per gram tutes 4 ee
in lactese |
20° C. 31a bile. |Wet basis. Dry basis.

1912 Dozen.
4279 D1 | May 550, 000 500, 000 AtO00 sess 34 Fences PS RIE | LOE eh Fy 15
4576 D3 | June 20 400 400 10 0.0017 | 0.0065 73. 85 6
4581 D3 jac 10 ee 13,000 3,000 1,000 - 0017 . 0063 72. 87 6
4634 E 4 | June 28 700, 000 3,300 1,000 . 0019 . 0068 72.18 6
4947 E6 |Aug. 9 49,000 (4) 10, 000 . 0023 . 0082 71. 81 6
41087 EZ Aug. 30] 1,600,000 1, 700, 000 10 - 0023 - 0084 72. 74 6

1 Less than 1,000.

The bacterial contents were widely divergent; the minimum num-

ber was 400 and the maximum number 1,600,000 per gram at 20° C.
The number of B. coli varied from 10 to 10,000 in the six samples.
The amount of loosely bound nitrogen was markedly higher in the
August samples than in the June samples.
Sample 41087 represents about the lowest quality of dirty eggs used
by reputable breakers for food purposes. The shells were so filthy
that dirty “ dirties” (the trade designation) best describes them.
The candle showed that they were shrunken eggs, and that many
yolks floated near the shell as if they were about ready to adhere to
it. The eggs eliminated during the process of opening were highly
infected, as follows: One egg with a green white, one sour egg, one
egg with yolk nearly mingled with white, and two eggs each with a
broken yolk which had at one time been adherent to the shell. The
conditions found in this lot of eggs are typical of low grade fall
receipts.
The bacteria found in this series of samples were, without doubt,
referable both to contamination from the shells during the process
of opening and to the eggs themselves.
EGGS HAVING CRACKED SHELLS.

On account of the heavy losses accruing from shipping eggs having

cracked shells, these eggs constitute one of the important classes
used for breaking purposes,
During the interval between May 2 and August 26, 1912, 2 sam-
ples of whites, 2 of yolks, and 16 of whole eggs were taken. These
samples represented lots of from 6 to 30 dozen cracked eggs, pro-
cured in D, E, and F houses, where it was the custom to keep
“checks” in cool surroundings from the time of receipt until the
time of breakage, and to give them precedence over other eggs in
regard to promptness in candling and breaking. The laboratory —
results (see Table 17) from these samples indicate what is to be
 COMMERCIAL EGGS IN THE CENTRAL WEST. 45

expected from cracked eggs that are broken in the first concentrating
center.
TABLE 17.—Hggs having cracked shells.

WHOLE EGGS.

oo
4&3 | Percentage
Total number of |&"2 ofammo-
bacteria per gram |3-4 niacal ni- ;
: on plain agar in- |2 & trogen,| &
8 cubated at— rar Folin 5
Bs) Sh mound es 2 Remarks.
& pal | S
ra) 2 i OY 8 3 rs) §
B = Bee| 4 |a 2 &
do) Boe =Q
po! & ° e . Q 15)® Ko} sg i) =

Z
=
oa)
gS
A
sess]
1)
as Z
a
2S ea)m4
fan)
o
asA
1912. |
4215 Fil) May 2 600,000; 600,000 0)0. 0017/0. 0056) 70.28] 15 dozen. .
4216 IP WS Clon ee 600 1,000 0} .0014] .0046) 70.98}...do....-..
4242 Fi} May 3 500, 000 3800, 600/10, O00}......].....-)------ 11 dozen. -
4254 D1} May 6 750, 000 490, 000 100} . 0016} .0052] 69.14] 15 dozen. .
4314 E 2} May 14 230, 000 80, 000/10, 000)......)..-..-|------ 12 dozen. -
4557 D 3| June 17 340, 000 120, 000}10, 000} . 0015] . 0052) 71. 20) 30 pounds.
4572 D 3| June 19 140, 000 75, 000 0} .0019] .0071| 73.22) 6 dozen...
4597 E 4| June 24 460,000; 330,060 NO) ieeeees Ep Me Ee 132 dozen. .
4635 E 5} June 28 2, 800 800 0} .0019] .0070| 72.82} 6 dozen. - .|
A711 D 4| July. 10 700 400 Qe eases te al i 15 dozen. .
4827 F5 July* 24 | 2,200,000} 1, 800, 000 10] .0021] .0C68) 69. 62)...do..-....
4963 F 6} Aug. 13 35, 000 35, 000 10} .0018] .0065) 72.51)...do....-..
4965 bP Bel doe e | 2. 700,000) 2,400, 000| 1,000\.2 <2 2 | ee edo
4993 F 6) Aug. 16 50, 000 38, 500} 1,0C0} . 0018] .0065) 72.29)...do.-....
4994 18 Gis cClossse 190, 000 TAO OOO SOO Se AAs
Ge sees eG ovata
41062 E 7| Aug. 26 | 2,300,000 950, 00C} 1,000} .0023) .0074) 69.01) 30 dozen. .

WHITES AND YOLKS.

4218 Fil| May 2 36, 500 1, 400 100)0. cru, 0016] 87.36) 30 pounds.| Whites. i;
4219 Waleedo 6, 800 ZOO 100 (ee
eee ali doen Yolks of No. 4218.
4282 2 1] May 8 13, 500 US OOO) COW easceslasseosesssce 25 pounds.| Whites.
4283 ies -do.... 25, 000 64, 000 yo ears ae |eas | MAG ON Yolks of No. 4282.
| |

Although the variation, shown in Table 17, in the number of bac-

teria in the different lots of cracked eggs was almost the same as
that found in the samples of seconds, the individual counts in many
instances were higher. For instance, 43.8 per cent of the samples
of checks, as compared with 66.6 per cent of the samples of seconds,
had counts under 200,000 per gram. The damaged shell of the
checks offers less resistance to bacterial invasion than do sound
shells, which factor explains the higher average count of cracked
eggs. As was the case with the types of eggs previously discussed,
the samples with the highest bacterial content were those which were
taken during the month of August. The samples showed no greater
deterioration chemically than did the specimens of summer firsts,
seconds, and dirties.
Eggs with incipient odors but normal in appearance are very
often found in some lots of cracked eggs, particularly in those which
have not been chilled previous to opening. An examination of four
single eggs gave counts of 12,000 bacteria per gram or less in three
samples and of 1,200,000 in the fourth. There was no abnormality
in the latter to distinguish it from the other three. A fifth sample
46 BULLETIN 51, U. S. DEPARTMENT OF AGRICULTURE.

composed of five eggs with a similar but not strong odor contained
550,000 bacteria per gram and showed the low amount cf ammoniacal
nitrogen of 0.0016 per cent on the wet basis. Further tests are
necessary before it can be positively stated whether the faint odor-
of warm cracked eggs is due to bacterial action or to absorption from
surrounding materials. Experienced egg breakers recognize the
characteristic odor of warm checks and do not discard them.
Quite different were the results of eh examination of two indi-
vidual eggs, the odor of which was suggestive of the products of
bacterial growth. The eggs, normal in appearance, differed from
each other only in the intensity of the odor. The count of the egg
with the faint odor was 12,000,000, of the one with the strong odor
150,000,000. These eggs would be detected by careful grading and
eliminated from a food product.
On account of the damaged shell, checks in a short time become
infested with bacteria or molds or both. This result is hastened
when the shells are soiled with the leakage from other eggs. From
different lots of cracked eggs there were selected six samples consist-
ing of from4 to 12 eggs with moldy shelis. No eggs were included
unless the contents were normal in appearance, taste, and odor.
Table 18 shows that there was a count of 160,000 in one sample and
of from 1,200,000 to 12,000,000 in the other five. These results prove
that even if the contents of cracked eggs with moldy shells smell, —
taste, and look good the egg substance is infected with bacteria as well Ef
as with molds.
TABLE 18.—Cracked eggs with moldy sheils opened commercially in packing |
house.

' Total number of Num- | | Percentage

[ bacteriaper | ber of | Gelatin CAG |
| | gramcuplan (88S-PTO) | jigne
Geiauin | ¢ai nitro- Per |
Sam- | Date of | agar at— | ducme |)~o |B eM, Folw |cent | ar boos
g
le | Source.| collec-
ae
| bacteria}
per
“7278
organ-
|| method. | of |
Sed sample.
Satis
tion.
| l gram in Sarthe “ture.
20°C. | 37¢¢. |lactose gr Wet | Dry
ee) bile basis.| basis
Baro a. | aie Beer J
| | 1912 |
4217 | Fij May 2 |1,2 200, 000/11,
300, 0600) 10) 230, 000/9. 0019/0. 0052) 69.31) 8 eggs..| Odor and 1
taste
| | | | good i}
4241| F 1) May 3 | 2,700,000, 700, 000 10} _15,000] .0013] .0043| €8.26 12 eggs. Do.
4398 D 2} May 27 4, 050, 0600/3, 199, 000: 10C)0in 1, 000) . C019} .0058) 67.17 S eggs..| Odor good.
4583} D3) June 20 | S. 60, 000)5, 490, 000, 100} 49.0C0} .0021| .0070| 70.18| 4eggs.. Do.
epee E 4; June 24 /12, 000, 000/5, 900, 000! 0 0) .0025 | (ily (1 Bese Do.
D6 | Aug. 20 | 160,000, 140, 000) yw eee 0019} . 0064 70.23 Geggs..- Do.
|
MOLDY CRACKED EGGS COMPARED WITH NONMOLDY CRACKED EGGS.

41036 D 6} Aug. 21 loo600, 000'13,000,000} 100, 000)......... 0. ee OO7TS} 70. 3g 11 Ibs... Malcy
shells. |
41037) D ‘|Aug. 2 2; 700; OOO'1; 700; 000} 2-222.
1s Pee 0022 . 0079} 72. 09 48 Ibs...| Nonmoldy |
| shells
from.
| | same lo
| } | J] | as 41036.
Pa is } | A
Kiara! t
 COMMERCIAL EGGS IN THE CENTRAL WEST. 47

On one occasion a few cases of checks, a large percentage of

which had moldy shells, were received at D house. The eggs for
breaking were separated into two parts, one consisting of about 13
dozen moldy eggs and the other of about 60 dozen nonmoldy eggs.
Every effort was made to eliminate all eggs abnormal in appear-
ance or odor. On. account of the number of times that it was neces-
sary to wash hands and to change apparatus after bad eggs, it was
a laborious task to separate the good from the bad. For example,
there were 3 eggs with a bad odor, 4 rots, 5 sour eggs, and 14 eggs
with a green white that were eliminated from the moldy lot. There
were, as given in Table 18, 2,700,000 organisms in the nonmoldy and
22,000,000 in the moldy eggs. The quantity of ammoniacal nitrogen
was practically the same in the two specimens, and was no higher
than that found in some samples of summer firsts, seconds, and
dirties. These results show that either the bacteria had not been
present long enough or had not multiplied to a sufficient extent to
materially change the composition of the egg substance.
The laboratory data from both the large and small samples show
that for the preparation of a food product with a low bacterial
content cracked eggs with moldy shells should be omitted.
EGGS HAVING THE YOLKS PARTIALLY MIXED WITH THE WHITES.

The class of eggs having the yolks partially mixed with the
whites consists of three forms: One in which the yolk has seeped
through the vitelline membrane in sufficient quantity to give a
yellow tinge to the thick portion of the albumen; the second in
which the yolk streams through small openings in the membrane
into the white; and the third in which the yolk membrane breaks
and allows the yolk to flow into the albumen. The first form can
not be detected by the candling process; the second and third in
many cases can be recognized by this method. During warm
weather the vitelline membrane becomes thin and the white less
viscous, which condition, accompanied by the jars occurring during
transportation, causes the membrane to break and the contents of
ihe yolk to escape. The cause of the seeping yolk in eggs is not
understood. The three forms are the predecessors of the class of
eggs called white rots, or eggs with white and yolk entirely mixed.
During the season of 1912, 10 samples, consisting of from 1 to 8
eggs, with yolks seeping into white were taken. The odor and taste
of each were good. The results are shown in Table 19.
48 BULLETIN 51, U. S. DEPARTMENT OF AGRICULTURE.
TABLE 19.—Small samples of eggs with the yolks beginning to seep into the
whites.
| | |

Total number of bac- | __ | Percentage of

teria per gram on |Number! q,jaiin | ammoniacal
i plain agar incuba-| Ofgas- liquefy- | Nitrogen, Fo- Per- | ne
Sample Date oi ted at— producing ing organ- lin method. cent- | Size of
NG Source.| collec- bacteria cane per | age of | sam-
E SIONS... i=
Deena
ae anc. -| de ee See
3 z in lactose g 5 Wet | Dry ture.
20 C: 37 Ce bile. basis. basis.

1912 | | 2ggs.
4194 E lj Apr. 25 2,600 3, 900 0}: 0:30: 10; O00l¢ s2: sie SE eee 3
4251 D1) May 6 67, 000 21,000 100] 0in 10,000} 0.0017} 0.0062) 72.61) 8
4266 D1) May 7 100 400 O)iestics 2. eles oe 23ers. se eee Se 1
4290) D1) May 91} 22,000,000) 21,000,000 0} 0in 10,000) .0021) .0077| 72.58 5
4357 E 2) May 17 100, 000) 21,000 10 900; O00) 2 S323. Sse ay elee 2
4406 D 2} May 27 44,000 5, 000 0} 0in 10,000} .0020) .0076) 73.76 5
4428 D 2} May 28 300 100 0) aosck se sees Eo a el ae 2
4895 D 5} Aug. 2 3,900,000) 3,500,000) Oin1@0)._--.-..-..- . 0023} .0078) 70.54 5
41001 D6) cAue.19*)<Ocin £05000)0:2225.105,000) 420 in (OO): eo a ee oe | 1
41032 D 6) Aug. 21 ie | 7, 500 TA) ets spe a" SODIS As: oe 74. au 4

Table 19 shows that the number of bacteria varied from 100 per
gram at 20° C. to 100,000 in eight samples and from 3,900,000 to
22,000,000 in the other two. No explanation can be given of the
cause of the last two counts other than that they may represent
iransitional stages between yolks beginning to mix and those entirely
mixed with white. It was shown on page 30 that the latter are
heavily infected with organisms. There were no B. coli in one-half
of the specimens and only a few in the remaining half. The amount
of protein deterioration was no greater than that found in seconds
and cracked eggs.
Only three samples were taken of eggs with the yolk entering the
white through small apertures in the vitelline membrane. The speci-
mens consisted of individual eggs and were practically sterile.
During the latter part of July two samples were taken, consisting,
in one case, of 30 dozen eggs, and in the other of 11 dozen, which,
before the candle, appeared to have broken yolks but were other-
wise normal. On opening the eggs it was found that the grading
by the candle was not accurate, because among them was a large
number of eggs which were approaching the stage of white rots. The
large sample was opened without much care in grading; the smaller
one was broken, and every ege.which appeared to have passed the
first stage of physical degeneration was eliminated. The difference
between the laboratory results of the two samples was striking.- The
poorly graded sample contained 8,300,000 bacteria per gram, 1,000,000
B. coli per gram, and 0.0028 per cent of ammoniacal nitrogen on
the wet basis; the well graded one, 5,500 organisms per gram, 10
B. coli, and 0.0021 per cent of loosely bound nitrogen. This experi-
ment is a good example of the effect of careful and intelligent grad-
ing on a product prepared from doubtful eggs.
These preliminary studies indicate that eggs with yolks showing
the first signs of deterioration are suitable for food purposes, if
 COMMERCIAL EGGS IN THE CENTRAL WEST. 49

other characteristics are normal, but that egos showing more advanced
decomposition should be avoided. This phase of the egg problem will
be specially investigated during the season of 1913.
BLOODY WHITES.

Many of the first eggs laid by pullets contain blood which may
be diffused through the white or may be in the form of clots on the ~
yolk or in the albumen (see Pl. IV). It is probable that in the
passage of the white and yolk through the oviduct some of the smal!
blood vessels are ruptured, thereby allowing blood to gain access to
the white or yolk before they are incased in a shell.
The examination of six individual eggs of this type disclosed the
presence of less than 5,000 organisms per gram. No &. coli were
found, except in one sample, which had only 10. The eggs were
taken from the cups of the breakers; consequently the few organisms
found are very likely referable to outside sources and not to the eggs.
On account of the presence of blood, these eggs are not used for food
purposes.
; EGGS WiTH BLOOD RINGS.

Eggs showing signs of incubation were discussed quite fully on

page 13; consequently only the information gained when these eggs
were broken commercially will be given here.
During May, 1912, four samples, consisting of from 3 to 12 large
blood rings, were taken. Since the weather had not been warm
_ enough to cause spontaneous development of embryos of fertile eggs,
these blood rings were probably the result of the undesirable practice
of selling incubated eggs which would not hatch. The yolks were
broken and partially mixed with albumen; the germinal disk was
deteriorated. The odor of each was good.
TABLE 20.—Eggs with blood rings.
LARGE SAMPLES OF EGGS WITH SMALL BLOOD RINGS.

2g Percentage
a 7 = .
Number of bacteria 3 oe . aeaee 2
per gram ai— as | AE |
hic Bens ee Z
Sample
amp Date
at of f See
“SDae CENCE 1S)S Size of -E
No. collec- pS e | ea —| = | sample. Remarks.
tion. oy Z a
A S Seo gis a ses
:
S
R
2S
AQ
;
tt
ar) Z
6«|BBS|
oe, =
8 | eB | gs
aD
a
<
a

1912. |
4837 F 5} July 25 36, 500 3,500 100/9. 0022)0. 0077; 71.50} 30 dozen..| Kept in chill room
2 weeks.
~ 4843 F 5) July 26 37, 000 37, 000 10} .0022) .0077| 71.60) 12 dozen...
A883 | D5] July 31 50, 000 41,000). .--.-- .0019} .0067| 71.56) 83 pounds.
4889 | D5) Aug. 1 /10in 1, 000/10 in 1,000 0| .0018} .0063) 71.58) 4 pounds. .
4960 F 6) Aug. 12 950,000) 700,000) 10,090} .0023) .0074) 68. 89) 20 dozen...
4975 F 6} Aug. J4 430,080} 500,000 10] .0024} .0072) 66.58)...do.....-
41033 | D6) Aug. 21 77, 000 58, 000 €} .0019 ales 74.59) 43 pounds.
i }
a i Less than 1,000.
17625°—14——__4
50 BULLETIN 51, U. S. DEPARTMENT OF AGRICULTURE.
TABLE 20.—EHggs with blood rings—Continued.

SMALL SAMPLES OF EGGS WITH LARGE BLOOD RINGS.

2= Percentage
Number of bacteria |S ohare? S
: aH = .

per gram at— a B) gen, Folin ioe

n 5 , n

Sample
Date of SeS=| method i) :
Size ofzs
No.
collec-
aril
ola
gg Z es zi =
= sample.
Remarks.

a
5) S =§
ESO
as =
Ee
a) g
2 5 0 ELEN POM | Seren ates
oo R 3 A = A a

1912. ;
4292 D1} May 9 1,700 400 (7) RSC ee Sige eee 3 eggs....- Broken yolks.
440] D 2} May 27 100 100 0\0. 0014|0. 0052] 72.97; 12 eggs... Do.
4402 ID WAS aOR Se a 400 790 10} .0016| .0059} 72.71]...do...... Do.
4430 D 2} May 28 100 150 0; .0019) .0070) 72.71) 5 eggs. -.--- Do.
4699 D4 July 9 | 7,100,000 400 0} .0018) .0065}) 72.09) 8 eggs..-_.. Do.
mt

LARGE SAMPLES OF EGGS WITH LARGE BLOOD RINGS.

4838 F 5| July 25 |} 4,000,000 1, 900, 0001100, 000lo.ao22lo. 0077| 71.41} 30 dozen..| Kept neo room
2 weeks.
4844 F 5} July 26 | 4,300, 000} 3, 100, 600 100} .0019} .0068} 72.17) 9 dozen...
A884 D 5; July 31 |0in1,000| 0in i1,000)....... . C019) .0063; 70.00] 74 pounds.
4888 D5) Aug. 1 6,500 | 0 in 1.000 10) .0919} . 0063} 69. 96} 10 pounds.
41040 D 6} Aug. 22 | 2,000,000} 1,400,000} 10.000) . 0020) .0071} 71.98) 7 pounds. -

The results of the first four samples given in the second section of
Table 20 showed very few organisms and no B. coli except in one
sample. The amount of ammoniacal nitrogen was identical with that
found in contemporaneous samples of seconds, cracked, and dirty
eggs. <A fifth sample, taken in July, consisting of eight large blood
rings, gave a count of 7,100,000 bacteria per gram on agar plates
incubated at 20° C. and of 400 on similar plates kept at 37° C. The
divergence of the two counts is not explained.
In July and August larger lots of both small and large blood rings
were studied. These were caused in part by the warmth of the late
summer months and in part by a short period of incubation under
broody hens. The first observations were made of a case of blood
rings which had been held in a chill room at about 32° F. for two
weeks. The eggs were recandled and 83 dozen eggs with broken yolks
or with yolks stuck to the shell were discarded. During the process of
breaking, the small and large blood rings were separated, the basis
cf division being less than 8 centimeters for the small and over ©
that for the large blood rings. Many of the eggs of the former type
had firm whites and yolks with faded rings. The eggs with the large
blood rings contained broken yolks. All eggs showing signs of mix- |
ing of white and yolk, often termed “runny eggs,” were excluded. — |

Each lot was mixed thoroughly by passing a few times through a

|
steamed sieve. The resulting mixtures had a good odor. The sam-
ple of small blood rings contained 86,500 bacteria per gram; the one
 COMMERCIAL EGGS IN THE CENTRAL WEST. o1

of larger blood rings, 4,000,000. The amount of ammoniacal nitro-

gen was in each case 0.0022 per cent on the wet basis.
The counts of similar samples of eggs with blood rings which iad
not been kept for any extended period in a chill room varied from less
than 1,000 to 950,000 for small blood rings and from under 1,000 to
4,300,000 for large blood rings. Since the blood rings which were
used in these samples underwent the same diversity of conditions
before and during marketing as did the seconds, cracked eggs, dirty
eggs, etc., they would be expected to show practically the same varia-
tions in bacterial contents.
The amount of loosely bound nitrogen in the samples of both small
and large blood rings was no greater, and in many cases was less,
than that found in stale eggs. These results are in accordance with
those found by Pennington and Robertson,’ which they summarize
as follows:
The amount of loosely bound nitrogen in incubated eggs, as determined by
the Folin method, shows an interesting change. In the case of infertile eggs
a very noticeable and quite regular increase takes place with time, while in
the case of the fertile eggs the increase is very slight. Considering the content
of loosely bound nitrogen as a criterion of protein decomposition, this is not
surprising, since in the first case heat would be expected to increase catabolic
processes, making for simpler nitrogen compounds, while in the second case it
introduces metabolic or upbuilding processes,
The product obtained from eggs containing small bined rings was
normal in appearance, taste, and odor; that obtained from eggs with
large blood rings. had a much lighter appearance; it was normal
with respect to odor but had a flat, insipid, and uninviting flavor.
Tt is evident, therefore, that changes had occurred which were re-
corded by the senses and not by the examinations just described.
Pennington and Robertson found that catalase increased in fertile
eggs during incubation, but did not increase in nonfertile eggs held
under the same conditions. It may be that the studies now under
way on the sugar content of eggs will also throw some light on the
changes occurring in fertile eggs during the process of incubation.

EGGS WITH TURBIDITY IN THICK WHITE.

It is observed that when eggs which have been in storage for some
time are broken, many of them have a turbidity which is localized in
the thick portion of the albumen, but this cloudiness disappears
_ when the eggs are warm. That this is a physical change brought
| about by low temperatures, and not by bacterial action, is indicated by
| the results given in Table 21 of the laboratory examination of six
samples of eggs with cloudiness in the thick white.
1U. S. Dept. of Agr., Bureau of Chemistry Cir. 104.
52 BULLETIN 51, U. S. DEPARTMENT OF AGRICULTURE.

TABLE 21.—Hggs with turbidity in thick white.

INDIVIDUAL EGGS.

|
Total number of | |
bacteria per |Number) qajiin
gram on plain | Of gas- |liquefy-
Date of agar in producing} ~. a:
eacuple Source.| collec- ape eer es bacteria Bes oo See ot Description.
PR tion. per gram 8 SAEED:
=aaa ae lactose ee
bile. Stat.
20° C. Bile C-

1912. | Pounds
4571 D3 | June 18 150 650 Oks reece
he eee Leaking sheil; normal
| yolk; no odor.
4575 D3 | June 19 43,500 36, 500 100 250008 Essen aa Do.
4587 D3 |June 22 150 100 0 igre aes Opened aseptically.

SMALL SAMPLES.

4598 E4| June 24 |120, 000 32,006 10 | 100,000 = Cracked eggs; normal
yolk; good odor.
4611 E4 | June 25 16,000 500 O fecdie cseee 1.5 | Cracked eggs and sec-
| onds; good odor.
4624 E4| June 27 290, 000 14, 000 10 10, 600 1 Dirty eggs; good odor.
}

EGGS HAVING WHITE PARTIALLY COAGULATED BY BEAT.

Breaking-stock eggs are occasionally found with contents which

present the appearance of soft-boiled eggs... They probably had been
dipped in hot water to prevent their use for hatching when they
had been purchased, ostensibly for food purposes. The heat of sum-
mer is also, under some circumstances, sufficiently great to cause the
albumen of eggs to partially coagulate, thus giving it a clouded ap-
pearance. For instance, an egg laid on a haystack exposed to the
direct rays of the sun becomes partially cooked and has the ap-
pearance of an egg which has been in boiling water about a minute.
A bacterial examination of two such eggs showed them to be prac-
tically free from organisms.

EGGS HAVING ENTIRE WHITE TURBID.

Bacterial growth in an egg may cause cloudiness in the albumen

analogous to that caused by the growth of bacteria in laboratory
media. ‘The eggs mayor may not have an odor. Eggs with a charac-
teristic sour odor (see p. 61) have, almost invariably, a turbid al-
bumen. An examination of four eggs with a clouded white showed
(Table 22) that their bacterial content varied between 15,000 and
150,000,000 per gram. The two eggs with the high counts had an
abnormal odor, which fact was indicative of the presence of large
numbers of bacteria.
 COMMERCIAL EGGS IN THE CENTRAL WEST. 53

TABLE 22.—Eggs having entire white turbid.

[+ denotes presence.]

Total number of bac- | Number

teria per gram on of gas- A
in i siete Gelatin
Sample
N Source.|e DEN CL
collec- pip pearsncubated peoduents
bacteria liquefying
IGT Se ank
Description.
G: tion. per gram Seas.
in lactose Perse
20° C. BU? Oy bile.

1911.
533 F¥ | Aug. 41] 5,600,000 | 3,400,000 100, 000 + | Milky white; normal
yolk; no odor.
1912.
4260 Di1| May 7 | 18,000,000 | 8,300,000 100 0 in 10,000 Seal odor; normal
yolk.
4331 E 2} May 16 {150,000,000 |120, 000, 000 10,000 | + in 160,000 | Unpleasant odor; cloudy
rim of white around
yolk.
4766 E5 | July 17 15, 000 4, 800 LOO aoe sae: - A very cloudy white
odor and taste good.

WHITE OR LIGHT ROTS.

White or light rots are the advanced forms of partly decomposed

egos, of which the following are typical: Eggs with yolk partially
mixed with white, eggs containing old broken-down blood rings, and
eges with a broken yolk which was previously adherent to the shell.
Before the candle these eges are light in appearance, hence their
mame, and are often passed as good eggs by candlers who do not
take the time tc determine the condition of yolks. Out of the shell
white rots appear as an unappetizing homogeneous mixture of yolk
-and albumen (see Pl. VIT).
During the spring of 1912 six samples, composed of from four to
eight white rots, were taken. The condition of these eggs had not
been detected by candling, and they therefore found their way to the
breaking room. Instead of being consigned to the rotten-egg bucket
they were poured from the cups of the breakers into sample bottles.
The laboratory examination showed that the majority of the samples
were heavily infected with bacteria, among which were many B. coli.
The results, which are given in Table 23, are in accordance with those
obtained in the study of white rots opened under aseptic conditions.
The percentage of ammoniacal nitrogen found in five of the six speci-
mens was greater than that found in any of the previous samples dis-
cussed. ‘The sampies of summer firsts, seconds, checks, and eggs with
yolk partially mixed with albumen contained from 0.0014 to 0.0026
per cent of loosely bound nitrogen on the wet basis, whereas the speci-
mens of white rots gave a variation of from 0.0019 to 0.0061 per cent
in the amount of ammoniacal nitrogen in five of the six samples
examined.
04 BULLETIN 51, U. S. DEPARTMENT OF AGRICULTURE.

TABLE 23.—Hggs having the white entirely mixed with the yolk.

Total number of bac- | Number Percentage of

varie per aoa ie of gas- Se Per
: plain agar incubated | producing| Gelatin nitrogen Size
bacteria | liquefying |Polin method. Coe of
No. |, Source.
Sample} oe 2 geo
pate?
per gram | organisms
ion in per gram, Sar se |e8© lises
poae
O! |sam-
OTS
pie.
20° C 37° C lactose Wet | Dry :
: : bile. basis. |basis.
| | J tt | | | | |__|
1912. 2 99
4261 Di|May 7 1, 100, 000 500, 000 10 | 0 in 10,000 |0. 0025 |0.0096 | 73.27 6
4291 Di1j|May 8 14, 000 10, 000 1,000 |0 in 10,000 | .0019 | .0072 | 73.63 5
4303 E2 | May 13] 82,000,000 | 60,000,000 | 1,000,000 | 37,000,000 | .0034 | .O01T1 | 69.34 4
4318 H2 {May 14 | 110,000,000 9)100, 000 1,000 | 22,000,000 | .0033 | .0109 | 69.68 6
4409 B 21 May 27) 16,000,000 | 5,000,000 10 |0 in 10,000 | .0027 | .0103 | 73.77 8
4459 3 | June 1| 56,000,000
June 1] 56,000,000 | | 24,000,000
24,000,000 || 1,000,000
1,000,000 |
| 15,000,000
15,000, 000 |
| .0061
.0061 |
| .0211
.0211 |
| 71.17
71.17 |..-.-..

On account of the large number of bacteria which white rots con-

tain it is very important that breaking stock be candled with suffi-
cient care to prevent these eggs from gaining access to the breaking
room.
EGGS HAVING THE YOLK ADHERENT TO THE SHELL.

Two different forms of summer eggs with adherent yolks, termed

commercially “heavy spots,” were found. One is caused by the
action of high atmospheric temperatures and the other by exposure
to damp surroundings. In some cases both factors contribute to the
same result. Both types occur most frequently during the summer
and autumn months.
The heavy spots caused by heat are found in the following stages:
First, in which the yolk is so lightly stuck to the shell membrane
that a slight jar sets it free (see Pl. VII) ; the second, in which the
yolk is adherent and broken; and the third, in which a very small
portion of yolk adheres to the shell, the rest being partially or entirely
mixed with albumen. In the case of an incubated fertile egg, it is
observed that the adherent surface of the yolk is the hatch spot
or blood ring. The first and second types are usually without odor;
the third may, or may not, have an odor, and is classed among the ©
white rots on account of the mixing of yolk and albumen.
The eggs with an adherent yolk, produced by moisture, present a
characteristic appearance before the candle. The portion of the
yolk coming in contact with the shell is dark and often black in ©
appearance. The yolk is much more opaque than normal. When
the contents of these eggs are emptied from the shell, a large portion
of the yolks cling to the shell. They have commonly a sour or
putrefactive odor. This type of egg, if held for a sufficient ett
of time, may develop mold spots on the yolk.
Three samples with yolk very slightly stuck to the shell were
selected by candling. When opened many of the yolks dropped out
whole and left no mark on the shell at the place of contact (see
 COMMERCIAL EGGS IN THE CENTRAL WEST, 55d

Pl. IV). During the breaking care was exercised to eliminate all
egos showing signs of mixing of white and yolk. For example, from
Sample 4842 one egg was discarded; from 4891 twenty-two eggs, and
from 4919 a larger number. The number of discards gives, perhaps,
an index to the bacterial contents, for, as shown by Table 24, Sample
4849 contained 900,000 organisms per gram; Sample 4891, 8,300,000;
Sample 4919, 24,000,000. The amount of ammoniacal nitrogen in the
first sample was practically the same as that found in August checks,
dirties, and seconds; the quantity in the other two samples was
slightly higher. The odor and taste of the liquid product was good
in every case. |
TABLE 24.—Small samples of eggs having the yolk adhering to the shell.

SLIGHTLY ADHERENT.

Number Percentage of
Number of bacteria | of gas- ammoniacal Per
per gram at— producing | Gelatin nitrogen,
Sample] gouree Dele ot bacteria |liquefying |Folin method. con Size of
No. 1 ‘tion per gram | organisms & oig-| Sample
in pergram.| _ ae
20° C 37° lactose Wet | Dry
x : : bile. basis. | basis.

1912.
4842 F5| July 26 900, 000 750, 000 HOO Aare
eae ae 0.0022) 0.0077] 71.37] 6% doz.
4891| D5|Aug. 1] 8,300,000) 5,800,000 Na eee Ss 0024} 0088} 72.72) 44 Ibs.
4919 E6 | Aug. 61] 24,000,000} 2,700,000 100)..... so ieee . 0024] .0086) 72.10) 1 Ib.

HEAVILY ADHERENT.

4422! D21| May 28} 30,000,000} 21,000,000 0| Oin 10,000] 0.0049] 0.0179| 72.69] 4 eggs.
4841 F 5} July 26 | 150,600,000} 120,000,000) 1,000, 000).........-. -0031} .0108) 71.25} 5 doz.

For comparative study two samples of eggs with yolks heavily

adherent to the shell and sometimes called by the trade “cellar stucks”
were selected by candling. The portion of the yolk which clung to
the shell when its contents were emptied was not removed. ‘The
product had an abnormal and offensive odor. Table 24 shows that
the bacterial counts, as well as the percentage of ammoniacal nitrogen,
| were higher than those found in the first type, where the yolks were
| slightly adherent to the shell. |
A sample of three eggs with adherent yolks, which had become
moldy, contained 7,200,000 bacteria per gram and 0.0022 per cent of
ammoniacal nitrogen. If this sample had included “moldy spots”
with offensive odors the laboratory results would show much more
decomposition.
Further studies will be made of spot eggs. The results cited, how-
ever, indicate that these eggs should not be used in a product pre-
pared for food purposes.
56 BULLETIN 51, U. S. DEPARTMENT OF AGRICULTURE.
BLACK ROTS.

Black rots receive their name from the black appearance which
they present before the candle. Out of the shell they are a homo-
geneous clive green liquid with an offensive odor resembling that
of hydrogen sulphid. It 1s often possible to pick out a black rot
from receipts by the odor and grayish tint of the shell. The results
of two samples consisting of from four to eight of these rots showed
counts of more than a billion bacteria and an amount of ammoniacal
nitrogen much greater than that found in any previous type of ege
discussed (see Table 25). Black rots, therefore, represent eggs in
the last stages of decomposition. ‘Their only value is for fertilizer
purposes.
TABLE 25.—Small samples of black rots.

e | Percentage
Number of bacteria per | Number of ammon-
gram at— R ofPRG Gelatin eat eae Per
f
ucing | liquefy- | Sen, Folin | cent | q.
Wate @ bacteria | ing or- | method. -| of | Size of
a Source.) collec- mois_| Simple.
Om) |asa ee ve ae pereram |eanisms
| in lac. |per gram. ture.
° ° tose bile. Wet | Dry
oe oe basis.| basis.

;
310,000,000) 10,000,000] 99,000,000)......|....-. 73.31] kebes:
1912.
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D2] May 28 | 1, 100,060,000]
D5) July 31 | 2,300,000, 000) 2,300, 000, 000) 10,000,000}. .-.----.- 0. 0229/0. 0800} 71. 40} 8 eggs.

DETERIORATED EGGS NOT DISTINGUISHABLE BY CANDLING.

The eggs previously discussed have been recognizable before the

candle or distinguishable by other characteristics, such as dirty or
cracked shells. Some eggs, however, which are distinctly undesir-
able, can not be detected by candling and must be eliminated by the
breaker. These are recognized when out of the shell by color, odor,
or general appearance. The great majority of them belong to three
groups: Eggs having a green white, often called “ grass” eggs by
the trade; eggs having a pungent, characteristic odor, commonly
known as “sour”; and eggs which are “ musty,” that is, having an
odor which is exceedingly penetrating, very characteristic, and oiten
suggesting that of the.common jimson weed. This odor increases
when heat is appled, so that a single musty egg in 100 pounds of
good egg will spoil it for bakers’ purposes.
A number of eggs have distinctive cdors when out of the shell, |
though there may be no visible signs of deterioration. Sometimes |
the eggs absorb these odors, such as the fruity odor which comes when
egos and apples are held in a closed space together, or that of straw-
board from the fillers in which they are packed. Unless these ab-
sorbed odors carry with them an objectionable taste there would seem ~
to be no reason for discarding the eggs. Other odors, apparently |
 *

COMMERCIAL EGGS IN THE CENTRAL WEST. 57

generated by the chemical changes accompanying deterioration, are

indications that the egg is unfit for food.
The various groups of eggs not recognizable before the candle are
productive of much trouble in the frozen and dried egg industry.
They will therefore be considered separately.
EGGS HAVING GREEN WHITES.

Certain eggs show a distinctly greenish tinge in the white (see

Pj. VIII). This may be so slight that 1t is not noticed unless com-
pared with an egg having a normal color. The majority of these eggs
show no other signs of deterioration; others have a thin albumen, a
yolk with a ruptured membrane or even mixed to a decided extent
with the white. Eggs showing macroscopic evidences of decomposi-
tion are usually accompanied by a fetid odor. The shells of eggs hav-
ing green whites are frequently cracked, stained, or dirty; many have
the appearance of washed eggs.
Tt is of interest to observe that these eggs are found in greatest
numbers in the spring, when dirty and wet shells are most prevalent.
Being but seldom distinguished by the candler while in the shell,
they go to the breakers. Some plants have in the past excluded such
eggs when the breaker happened to see them, especially if they had
reached the odor stage. Other cgi lacie: used them when odor-.
less and gave various reasons to account for the color, the most fre-
quent explanation being that the hen had been eating grass. This
supposition led to the term “ grass egg” as descriptive of this con-
dition, but as this phrase has been used by the trade to describe the
early spring eggs also, much confusion has resulted.
Laboratory examination disclosed the fact that such eggs contain
enormous numbers of bacteria, as 1s shown in Table 26.
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 60 BULLETIN 51, U. S. DEPARTMENT OF AGRICULTURE.

The samples given in Part 1 of Table 26 were opened aseptically.

The organisms found are, therefore, referable strictly to the egg.
The other samples were obtained in packing houses and were opened
into sterile cups. It is possible that a few extraneous organisms
may, therefore, be included in these bacterial counts, but the error
is small. There is a close agreement between maximum and mini-
mum counts in the samples obtained by the two methods. While but
31 individual commercial samples were examined, many of them rep-
resent a large number of eggs and a few approximate 5 pounds each.
Twenty-six of the 31 samples, or 83.9 per cent, show counts of over
10,000,000 per gram.
The predominating organism has been found to be Pseudomonas
syncyanea (Migula)* and the color of the egg white is due to the
ability of this form to produce a diffuse, green flucrescence in the
medium in which it grows. When pure cultures of this pseudo-
nonas were injected into a fresh egg the white assumed the char-
acteristic color in a few days and later developed a fetid odor.
The pseudomonas is not, however, in pure culture when occurring
in eggs with a green white. &. coli, as well as other organisms, are
generally found with it. The numbers of B. coli, as determined by
lactose bile fermentation, varied from 10 to 1,000,000 per gram.
The eggs which were physically in good condition and odorless,
and some of which were separated into white and yolk, are listed
in Table 26, Part IIT. Others, having an odor but not sufficient to
preclude use according to old methods of grading, are given in
Part TY. it will be observed that the white of the egg has a much
ereater number of organisms than the yolk, though the infection
in the latter is also extensive. A further indication of a mixed in-
fection is the fact that organisms which liquefy gelatin are com-
monly present in numbers. The pseudomonas isolated does not
liquefy gelatin. It does not grow to any extent at 37° C.; yet the
counts at this temperature are frequently decidedly higher than the
sum of the number of liquefiers and the organisms developing in lac-
tose bile with gas production. Apparently, therefore, these eggs with
green-colored whites are recognized by the characteristic color pro-
duced by one species, though they are the harbingers of a number
of species as well as of great numbers of organisms.
This argument is reenforced by the amount of loosely bound —
nitrogen found. When the egg is not separable into white and yolk
the amount of nitrogen is uniformly high—much higher than in
eggs commonly used for food. When the degeneration of the egg
is not sufficient to interfere with its physical integrity the amount —
of loosely bound nitrogen is not materially increased. It might ©
1This organism was identified by Evelyn Witmer, of the staff of the Food Research
Laboratory.
 COMMERCIAL EGGS IN THE CENTRAL WEST. 61

be inferred from these facts that even though the number of

bacteria in the egg be very high, as, for example, in Sample 4504,
where 210,000,000 per gram were found, the infection is too recent
to have produced chemical changes in the nitrogenous constitu-
ents. Because of the mixed infection it is not possible to correlate
the amount of loosely bound nitrogen with the presumably greater
or lower number of pseudomonas individuals, since the accompany-
ing organisms may exercise even greater activity in splitting protein
molecules.
SOUR EGGS.
The term “ sour eggs,” or “ sour rot,” is used by the egg breaker to
describe an egg that has when opened a peculiar pungent odor. In
the sense of a vinegar or common acid odor these eggs, in the earlier
stages at least, do not fit the name. In the later stages they may
have an odor suggesting sourness in the usual acceptance of the term.
They are characterized by causing a prickling sensation in the nose,
suggesting the bite of pepper, though not so sharp nor so well defined.
These eggs can not be distinguished by candling. Generally, how-
ever, there is some visible sign of degeneration as well as the charac-
teristic pungency. For example, sour eggs frequently have a tur-
bidity in the white; or the yolk membrane may be weak, or even
broken, so that the yolk is more or less mingled with the white. The
only means of detecting such an egg is the peculiar pungent odor.
Table 27 gives the bacterial and chemical analyses of 18 samples of
| sour eggs. The samples vary in size from 2 eggs to 5 pounds. All
were obtained from the current egg supply in the several packing
houses, were broken by cracking on a sterilized knife edge, and were
emptied into a sterilized glass cup. The grading was such that the
eggs in one lot were as nearly identical as possible.
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63
 \ ;

64 BULLETIN 51, U. S. DEPARTMENT OF AGRICULTURE.

All the samples examined show a high count of bacteria. One

sample (4269) has more than a billion organisms per gram and all
the samples but three have more than 100,000,000. On the score of
numbers of bacteria these eggs rank with black rots. The table
gives first a series of eggs in which the odor is but faint and which
might easily be passed by the careless or too rapid grader. Part I7
gives another series in which the odor was distinct and Part II
gives three samples in which the odor was pronounced. It will be
noted that these last three samples are individual eggs, and that a
physical deterioration in two of the three has proceeded so far that
the vitelline membrane has ruptured.
The bacterial content of the eggs with a faintly sour odor and those
with a distinctly sour odor is about the same, with the exception of
Sample 4256, in which the bacterial count agrees with the eggs in
Part IT in having a decidedly sour odor.
The great number of organisms invariably present is, however,
the noteworthy feature. Aside from the numbers of organisms, the
Bacillus colon was found in every sample examined, a condition
which, up to this time, has not been observed for any other single
type of deteriorated egg. Not only are B. coli present, but the num-
ber, as determined by lactose bile fermentation, is usually at least
a million, and may be 10 million. In some of the samples listed as
showing a million, more may have been present, because the dilutions
were not made beyond this point. Had they been it is quite probable
that the coli organisms would have been found to be more numerous
than the analyses indicated. One sample (41016) was separated into
whites and yolks. The count in the whites is double that in the yolks,
which may indicate an infection from the exterior, though more
work must be done with the two portions of the egg before accepting
this suggestion as a fact.
Where organisms hquefying gelatin were sought they were found
and in comparatively large numbers. Hence, there is in these eggs
a condition very much like that noted in the eggs with a green white,
namely, a mixed infection aggregating large numbers of individual
organisms and characterized by the presence of one distinguishing
species.
The amount of loosely bound nitrogen is higher than that com-
monly observed in seconds, which, on the average, is 0.0067 per cent
on the water-free basis. The range, omitting Sample 4256, which is
exceptionally high, is from 0.0102 to 0.0194 per cent, with an average
of 0.0184 per cent, all these values being on the water-free basis.
The water content of the samples varies from 69.52 to 75.48 per cent,
indicating a decided variation in the age of the eggs as measured by
shrinkage.
 COMMERCIAL EGGS IN THE CENTRAL WEST. 65

MUSTY EGGS.

The eggs called musty by the bakers have a strong odor, very
penetrating and persistent, becoming more pronounced when heat
is applied. All such eggs are sharply watched for by egg breakers
and discarded. Fortunately they are not very plentiful, even in the
early spring and late summer, when they are most common. Hot,
dry weather seems to lessen their frequency.
Sometimes several musty eggs will be found in the same lot; very
rarely almost a whole case of eggs will be of this type. They can
not be recognized by the candler and very frequently there is no
physical sign to indicate that the egg is not good. The sense of
smell alone must be depended upon to detect them.
The few examinations made of musty eggs do not justify any con-
clusions; therefore they are not given here. It is highly desirable
that further and detailed studies be made of this type of egg, which
is interesting from practical and scientific viewpoints.

. SUMMARY.
BACTERIOLOGICAL RESULTS OF INDIVIDUAL EGGS OPENED ASEPTI-
CALLY IN THE LABORATORY.

The first section of Table 28, summarizing the total bacterial con-
tents of individual eggs opened aseptically in the laboratory, shows
that the greatest percentage of second-grade food eggs examined,
the medium stale eggs, hatch-spot eggs, heavy rollers, dirty eggs,
cracked eggs, and eggs with yolk partially mixed with albumen, con-
tained less than 1,000 bacteria per gram. The occasional high bac-
terial content of single cracked eggs, dirty eggs, etc., could, in most
instances, be predicted by the appearance of the shell or by the odor
and condition of the contents. Such eggs would ordinarily be recog-
nized and discarded by the housewife or egg breaker.
The second section discloses the rather unexpected fact that B. coli
were not present in the whole-shelled second-grade eggs and were
present in only 5.9 per cent ot the cracked-shelled eggs.
Blood rings and the last five types of eggs given in the two sections
represent eggs ordinarily discarded as unfit for food purposes. The
first section shows that 26.5 per cent of the eggs with adherent yolks,
50 per cent of the eggs with dead embryos, 75.9 per cent of the moldy
eggs, 66.7 per cent of the white rots, and 100 per cent of the black rots
contained over 1,000 organisms per gram. A review of the second
section of the table shows that, with the exception of the white and
black rots, B. coli were present in but few of the eggs.
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68 BULLETIN 51, U. S. DEPARTMENT OF AGRICULTURE. |

BACTERIOLOGICAL AND CHEMICAL RESULTS OF COMPOSITE SAMPLES

OF EGG OPENED COMMERCIALLY IN THE PACKING HOUSE.

The laboratory results of composite samples of eggs opened com-

mercially in the packing house are summarized in Tables 29 and 30
and are shown graphically in figures 1 and 2. From these data the
following conclusions are drawn:
(1) The samples of July and August firsts contained very few
organisms, and in many cases no bacteria of the 2, coli group.

SUL ANO AUGUST FIRSTS!

SYALL “BLOOD RINGS.”

“SE.CONDS «
Co
DIRT £GES.
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Fic. 1.—Percentage of samples opened commercially with bacterial counts over 1,000,000
per gram,

(2) The majority of the samples of clean-shelled seconds had a

comparatively low bacterial content, only 8.3 per cent of them con-
taining over 1,000,000 organisms per gram. The number of B. coli
varied in the different specimens from none to 100,000 per gram.
(3) The percentage of bacterial counts over 1,000,000 per gram in
samples of dirties, checks, and eggs with yolk partially mixed with
albumen was 16.6, 18.8, and 20 per cent, respectively. No greater
number of B&B. colt was found in these samples than in samples of
seconds.
(4) The samples of blood rings contained comparatively few
organisms. The large blood rings in most instances showed more
infection than did the small rings. Most of the speeimens contained
less than 10 B. coli per gram.
 COMMERCIAL EGGS IN THE CENTRAL WEST. 69

(5) The amount of protein decomposition as shown by the am-

moniacal nitrogen in the preceding six types of eggs was greater, as
would be expected, than that found in strictly fresh eggs, but was
no greater than that found in some grocery eggs. Although a cracked

aici £6CS.

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Fic, 2.—Average amount of ammoniacal nitrogen in 14 types of eggs (closed bars, fresh
basis; open bars, dry basis).

er dirty shell may be a factor in facilitating infection and subse-

quent decomposition, the data obtained show that checks and dirties
in the producing section are as well preserved as the clean whole-
shelled seconds or the July and August firsts.
70 BULLETIN 51, U. S. DEPARTMENT OF AGRICULTURE.

(6) The eggs constituting the samples of July and August firsts,
seconds, dirties, and checks would be used without compunction by
the housewife, baker, or confectioner.
(7) The majority of the samples of white rots, eggs with yolk
lightly adherent to the shell, and all of the samples of sour eggs,
black rots, eggs with a green albumen, and eggs with yolk heavily
adherent to the shell, were infested with bacteria. 2B. coli were pres-
ent in most of these samples, forming the predominating organism
in the samples of sour eggs.
(8) The eggs with the yolk lightly adherent to the shell were, {

chemically, slightly lower in quality than were the second-grade food

eggs, whereas the sour eggs, white rots, eggs with a green white, and
eggs with yolk heavily adherent to the shell showed much more
deterioration. Black rots had five times as much ammoniacal nitro-
gen as any of these types of eggs. With the exception, possibly, of
the eggs with yolks lightly stuck to the shell, none of the eggs in
these samples would be used by the housewife or reputable baker or
confectioner.
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 COMMERCIAL EGGS IN THE CENTRAL WEST. | UB

TABLE 30.—Variation in amount of ammoniacal nitrogen in 17 types of eggs.

aN

Per cent of ammoniacal niiro-

Number gen, Folin method.
Kcind of eggs. of
samples
Wet basis. Dry basis.

TEINS ONSYR a a ae a a ya ee DAU yesHAP 6 | 0.0011-0.0015 0. 0040-0. 0054

Aibhlhy Qin! AVOUT ISU SS 5 bao eu coe saa do oes acouSadouocnoaosodue 4 -0019- .0022 -0065- . 0074
GOCCP ER OS oe ean ue oO es a eee A ee cet eee een 10 FOOUO= {OO 22 ese eee ee
Siuell CRCOS Bir ere eee net gers cca tehs Scueeintn= Siam ena teen 3 .0016- .0018 .0061— . 0069
SC COM CS rt eerie neem onnrete cepideyen a lieksSates aie Hence 17 .0015- . 0026 .0048- .0098
ETA CIRC ORG Sst agg PEN eal Ie a Ueepee eeiNenc eee nes ee a 18 .0014— .0024 . 0046- . 0083
Crackedlesssi with moldyrcheliss 7 eres ean see eee iG .0013- . 0025 .0043- . 0088
DT iygCCOSS See eH RRP ECE eR eee ee eee ie ee ae Te 14 .0013- .0024 | .0061- . 0084
Eggs with yolk partially mixed wath wihites. ©. 6° ost awe 5 .G017- .9023 .0062- . 0078
Simal blood anim gs. © jetta) ee ete ta ta Hr a ao hele ci Sie 7 .0018- . 0024 0063- .0077
CATOCHO] OO GMT E Sh ae Mee hoe yanNa yanipeiytee, Lal Eater eonL kteve 2 Se 0) .0014— . 0022 0052- .0077
Eggs with yolk slightly adherent to shell. ........-.....------ 3 -0022- .0024 0077- . 0088
Eggs with yolk heavily adherent to shell............-..-..--- 2 -0031- . 0049 0108- .0179
AVVAL GOST OLS se Spa Ain oi SS eR rN Sok a I ak jst cee OL Re peed 6 QO19- .0061 0072- .0211
Meosnwachereen\wiateses ace Meas ha eek oan eyes oes 12 0016= .0071 0056— .0264
SOUITCSCS es sires UN rau ea Sh voametat LI Se eR 12 0029- .0098 0102- .0323
IBIDCKSTOLSH emer ea: ook eee ena Re ee arse NR Tee 1 . 0229 . 0800

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A COMPARISON OF BACTERIAL CONTENTS OF INDIVIDUAL EGGS

OPENED ASEPTICALLY WITH THOSE OF EGGS OPENED COMMER-
CIALLY.

A comparison of the results of individual eggs opened aseptically

with the results of composite samples of eggs opened under clean
commercial conditions shows some apparent discrepancies. For in-
stance, only 4, or 7.1 per cent, of the 56 individual cracked eggs
opened aseptically contained over 1,000 organisms per gram, whereas
14, or 87.5 per cent, of the 16 composite samples, representing 2,924
“checks,” opened commercially contained more than this number per
gram.
Tt will be observed that the numbers of the latter are far in excess
of the former; it will be remembered, also, that eggs vary greatly
among themselves. It is possible, therefore, that the differences
between the bacterial findings of individual eggs and composite
samples are due, in large part, to the relative difficulty in detecting
early stages of infected eggs.
It was possible, for instance, to detect by the senses* but two of
the four individual cracked eggs which were infected. It has been
shown in Tables 26 and 27 that incipient sour eggs, which are de-
tected only by the sense of smell, and eggs with albumen just begin-
ning to turn green, which are recognized only by the sense of sight,
1To determine definitely to what extent it is possible to detect infected eggs by means
of the senses, and to what extent the bacteria! content of a product consisting of large
numbers of eggs of unknown history can be minimized by grading, it is necessary te make
detailed descriptions of the characteristics of many individual eggs, to open each aseptic-
ally and to determine their bacterial content singly and in combination. To find, also, the
amount of bacterial contamination acquired during the preparation, studies must be made
of the routine methods in use in egg-packing houses to determine the part which each step
in the process of preparation plays in the final condition of the product. This subject
will be presented as the second report of this series.
 74 BULLETIN 51, U. S. DEPARTMENT OF AGRICULTURE.

centain millions of organisms. It is reasonable to conclude, there-

fore, that the earlier forms of such and similar eggs furnish large
numbers of bacteria to the liquid product prepared from second-
grade food eggs. It is quite probable, also, that these earlier stages
of incipient sour eggs are a contributing cause to the presence of
appreciable numbers of B. coli in liquid egg of good quality.

TECHNIQUE FOR THE BACTERIGLOGICAL EXAMINATION OF

EGGS.
METHODS USED FOR OBTAINING SAMPLES OF INDIVIDUAL EGGS
OPENED ASEPTICALLY IN THE LABORATORY.

A. Mercuric chlorid method—The eggs were washed in running

water, treated for five minutes in a 1 to 500 or 1 to 1,000 mercuric
chlorid solution, and then rinsed with sterile water. The egg was
then placed, large end uppermost, in a suitable holder. A small open-
ing was made in the apex with sterile, fine-pointed forceps, about 2
square centimeters of the shell removed, and the membrane punc-
tured. About 2 cc of the white were then transferred with a sterile
pipette to a sterile tared weighing flask containing small pieces of
sterile glass. The opening was made larger and as much of the white
as possible removed with the pipette. Wuth a second sterile pipette
the vitelline membrane was ruptured, and about 2 cc of the yolk
_ transferred to another weighing flask. When it was impossible to
examine white and yolk separately, on account of disintegration, a
sample of whole egg was taken.
B. Flaming method.2—The egg was washed in running water,
rinsed in sterile water, dried with a sterile towel, and placed, large
end uppermost, in a suitable holder. The top of the egg was steril-
ized by flaming. A portion of the top was removed with fine-pointed
forceps and the contents of the egg dropped into a sterile salt-
mouthed 4-ounce bottle containing sterile glass.
PREPARATION OF COMPOSITE SAMPLES OF EGGS OPENED COMMER-
CIALLY IN THE PACKING HOUSE.

The details of the collection and handling of the samples are de-
scribed on page 39. When the samples arrived in the morning, they
were examined immediately. When they came late in the day, they
were put at once into a sharp freezer and held overnight. On ar-
rival at the laboratory the hard-frozen samples were placed imme-
diately in a water bath at 40° C. and allowed to remain, with frequent
shaking, until completely melted. After shaking the melted sample
vigorously for five minutes, about 3 ce were transferred with a sterile
pipette to a tared weighing flask.
1 Samples 4001 to 4159, inclusive, and 3001 to 3030, inclusive, were obtained by this
method.
2 Samples with numbers under 1,000 were obtained by this routine,
 COMMERCIAL EGGS IN THE CENTRAL WEST. (as)
METHOD OF PLATING AND COUNTING.

The flask containing the portion for examination was weighed.

_ The weight of egg material in grams multiplied by 9 gave the number
of cubic centimeters of sterile physiological salt solution required to
| make a 1 to 10 dilution, the shght error due to the gravity of the egg
' material being disregarded. After very thorough shaking, 1 cc of
| the 1 to 10 dilution was transferred by means of a sterile pipette to
9 ec of sterile physiological salt solution, thereby obtaining a 1 to
100 dilution. Higher dilutions were made on the same plan. Two
dupheate series of plates of nutrient agar were prepared from four
| consecutive dilutions. One set was incubated at 37° C. for two days;
the other for five days at 20° C.
The history of the sample was used as a basis for deciding in each
ease which dilutions should be plated. Whenever possible plates
containing from 50 to 250 colonies were selected for counting. The
numerical results were expressed in accordance with the rules pre-
scribed by American Public Health Association, 1912. A Stewart’s
counting chamber and a hand lens, magnifying four or five diameters,
were used to facilitate the counting. To determine the sterility of the
media, the salt solution, and glassware, blank plates were poured at
each plating. Plates of agar were also exposed to the air for three
minutes during each plating to show the relative freedom from air
contamination.
DETERMINATION OF THE NUMBER OF ORGANISMS CAPABLE OF LIQUEFYING GELATIN.

In a number of the experiments to be reported an attempt was

made to determine the gelatin liquefying organisms quantitatively.
For this purpose gelatin plates were prepared at the same time and
in the same manner as the agar plates. These were incubated at
20° C. and the liquefying colonies counted at the most appropriate
time. The counts were so discordant that many of the results were
discarded. The difficulty appeared to be due principally to the
fact that in most cases there was present a mixture of organisms,
some of which were capable of lquefying the entire plate in 48
hours while others required several days, or even weeks, to show the
first signs of liquefaction. The action of the slower ones was, there-
fore, masked by that of the more rapid.
After a large number of gelatin plates had been made and it was
found that irregular counts were very frequent, it was decided to
abandon the method except in special instances where special in-
formation was required.
DETERMINATION OF THE NUMBER OF ORGANISMS PRODUCING GAS FROM LACTOSE
IN THE PRESENCE OF BILE SALT.

At the time of plating 1 ce of each dilution was transferred to a

Durham fermentation tube containing lactose bile salt medium.
76 BULLETIN 51, U. S. DEPARTMENT OF AGRICULTURE.

These tubes were incubated for two days at 37° C., at the end of
which time each dilution showing gas was recorded as positive.
The fermentation tests were in every case started with the 1 to 10
dilution and carried at least one dilution higher than the plating.
The denomination of the highest dilution showing positive results
was reported as the number of gas-producing organisms in the
sample. This is the generally accepted presumptive test for B. coli
group.
FURTHER EXAMINATION OF THE GAS-PRODUCING ORGANISMS.

As far as time permitted, one of the higher dilutions from each

sample showing gas production was plated qualitatively either on
litmus lactose agar or Endo’s medium. From these plates typical
coli-like colonies were selected and examined to ascertain whether
they conformed to the definition of B. coli communis as given in
the 1905 Report of the American Public Health Association. For
this purpose they were subjected to the following tests, morphology,
Gram stain, motility, liquefaction of gelatin, coagulation of milk,
production of indol from peptone solution, reduction of nitrates to
nitrites, fermentation of lactose, and fermentation of dextrose.
They were then tested for gas production in dulcite, sucrose, man-
nite, and raflinose, in order to classify them according to the scheme
outlined in the 1912 Standard Methods of Water Analysis, American
Public Health Association.
CULTURE MEDIA USED.

The nutrient agar, gelatin, and broth were made from fresh beef
practically in accordance with the directions given in Standard
Methods of Water Analysis, American Public Health Association,
except that they were made in larger quantities than there specified
and were cleared with egg white and filtered through paper.
The lactose bile salt medium was prepared by dissolving 10 grams
of peptone, 5 grams of bile salt (commercial sodium taurocholate),
and 5 grams of sodium chlorid in 1 liter of distilled water, filtering
and adding 10 grams of lactose.
The sugar broths were prepared by adding 1 per cent of the sugar
to neutral sugar-free nutrient broth, made from fresh beef.
The milk was fresh, separator skimmed. It was used both with
and without litmus.

ANALYTICAL METHODS USED IN THE “EGG INVESTIGATION” DURING

THE SUMMER OF 1912, AT OMAHA, NEBR.

Ammoniacal nitrogen.—Briefly stated, the method consisted in

making the egg solution slightly alkaline with sodium carbonate,
driving out the ammoniacal nitrogen with a current of air, absorb-
| COMMERCIAL EGGS IN THE CENTRAL WEST. Le

ing the ammonia in standard sulphuric acid, and titrating the excess
of the latter with twentieth-normal ammonia solution, using congo
red as indicator. For liquid egg, 50 grams were weighed into a
liter suction flask, 200 ce of water were added, and the flask vio-
lently shaken until a uniform suspension was secured. There was
then added in turn 100 ce of 95 per cent alcohol, to prevent foaming,
2 grams of sodium carbonate, to render the solution alkaline, and 1
gram of sodium fluorid as a preservative, the mixture being shaken
after the addition of each reagent. The fiask was then put in place
and aspirated for five hours, using a strong air current. The latter
was first drawn through a 25 per cent solution of sulphuric acid to
remove ammonia, and was then delivered to the aspirating flask
through a tube with several small perforations, thus affecting an ex-
cellent distribution of the air current through the egg mixture. It
was then passed through a trap which caught any particles carried
over mechanically or through foaming, through the absorption fiask,
which contained 10 cc of twentieth-normal sulphuric acid diluted
with 50 cc of water, and finally through a second trap, which re-
tained any acid carried over mechanically. Three hundred cubic
centimeter Erlenmeyer flasks were used for the absorption apparatus
and traps. From time to time the flask containing the egg mixture
vas shaken to insure complete removal of the ammonia. After the
air current had passed for five hours the apparatus was disconnected,
the second trap and connecting tubes were rinsed off into the ab-
sorption flask, and the excess of acid titrated with twentieth-normal
ammonia solution, using congo red as an indicator. For desiccated
eggs 20 grams were weighed into a liter suction flask, 230 ce of
water added, and this same process followed.
Moisture—About 3 grams of egg were weighed into a small lead
dish (bottle cap), dried in a water-jacketed oven at 100° C. for one
hour, placed in a vacuum oven and dried in vacuo at 70° C. for 10
hours, cooled in a desiccator, and weighed. The sample was then
reheated in vacuo until constant or increased weight was noted,
weighing at intervals of two hours. The lowest weight obtained was
taken as the nearest approximation to the correct figure.
Ether extract—tThe lead dish and dried sample from the moisture
determination were cut into small pieces, placed in a Johnson ex-
traction apparatus, extracted for 16 hours with anhydrous ether, the
ether expelled on the steam bath and the fat dried at 100° C., cooled
in a desiccator, and weighed.

WASHINGTON : GOVERNMENT PRINTING OFFICE: 1914

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