Egg Storage and the Embryo1
G. M. Fasenko2
Department of Agricultural, Food and Nutritional Science, University of Alberta, Edmonton, Alberta, Canada T6G 2P5
ABSTRACT In domestic avian species, eggs are stored
at cool temperatures until they can be placed into an
incubator. The low temperature-induced diapause en-
ables the embryo to survive until optimal temperature
and humidity incubation conditions can be provided to
support embryonic growth. Egg storage is a logistical
necessity for the hatching egg industry both at the breeder
farm and at the hatchery. However, it is well known
that egg storage longer than 7 d negatively influences
hatchability. At the cellular level, long-term egg storage
induces cell death. This appears to be occurring both via
necrosis and apoptosis. The result is higher embryonic
mortality and, consequently, lower hatchability. In addi-
tion, long-term egg storage influences embryonic devel-
opment and metabolism. Embryos of eggs stored long-
term can be affected such that they do not initiate growth
Key words: fertile egg storage, embryonic development, embryonic viability, hatchability
INTRODUCTION
Arguably, one of the most important aspects of the
broiler production chain is the production of fertile hatch-
ing eggs. Hatching eggs for the broiler industry are pro-
duced on broiler breeder farms where males and females
are housed together. At most of these modern farms, the
hens lay eggs into nest boxes, the eggs roll onto mechani-
cal belts, and they are then transported to an egg-handling
location just outside the breeder barn. The eggs are col-
lected numerous times during the day to prevent the
fertile egg from becoming broken or contaminated with
bacteria. Upon egg collection, the eggs are placed into
coolers set at approximately 15 to 20°C and 75 to 80%
humidity. The eggs may be washed or sanitized postcol-
lection then stored in these on-farm coolers for approxi-
mately 3 d. Transportation of the eggs on specially envi-
ronmentally regulated trucks does not occur on a daily
basis, because most hatching egg farms are at consider-
©2007 Poultry Science Association Inc.
Received September 19, 2006.
Accepted October 7, 2006.
1
Presented as part of the Embryo Symposium: Managing the Embryo
for Performance, July 19, 2006, at the Poultry Science Association Annual
Meeting, Edmonton, Alberta, Canada.
2
Corresponding author: gaylene.fasenko@ualberta.ca
1020
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after proper incubation temperatures are provided; they
initiate growth, but grow at a slower rate than eggs stored
short term; and they are affected in both of the previously
mentioned ways. Development of equipment to measure
the embryonic metabolism of individually incubating
eggs over the entire 21 d of incubation has provided
further evidence that embryo metabolism has changed
due to storage. One of the methods to reduce the negative
effects of long-term storage has been to incubate eggs for
short periods before storage. In both turkey and chicken
eggs, this technique has been successful in improving the
hatchability of long-term stored eggs. It is hypothesized
that particular embryonic developmental stages are better
able to survive long-term storage. Future research should
focus on the mechanisms behind this improved ability to
survive storage.
2007 Poultry Science 86:1020–1024
able distances from one another as well as from the hatch-
ery; thus, daily egg pick-up would be economically inef-
ficient.
The eggs are transported to a commercial hatchery
where they are again placed in large coolers that have
temperature and humidity regulated. The length of time
the eggs spend in these hatchery coolers varies depending
on the current situation in the broiler industry and at the
hatchery. If there is a high demand for broiler chicks, the
hatchery manager will set the eggs fairly quickly. If there
is a surplus of hatching eggs, the eggs will sit in the
cooler for a longer period. In addition to considering
these factors, the hatchery manager must also manage
the number of eggs in large incubators. Because most
incubators in modern commercial hatcheries are large
multistage incubators, the efficiency of the incubator op-
eration is affected by the number of eggs set. Thus, the
hatchery manager must also balance egg numbers such
that the incubators are set close to capacity. Another factor
to consider is that in areas where high environmental
temperatures occur during the summer months, egg pro-
duction may be negatively affected. Hatchery managers
may anticipate this and stockpile eggs in the hatchery
coolers in times of plenty to prepare for anticipated lower
egg numbers. It is common knowledge in the poultry
industry that cool egg storage longer than 1 wk negatively
affects hatchability.
 EMBRYO SYMPOSIUM
A discussion of the effects of egg storage before incuba-
tion would be incomplete without a review of early em-
bryonic development. This paper will provide a general
review of embryonic development as it relates to storage,
as well as discuss selected research on length of storage
and the effects of storage on embryonic development
and hatchability.
EARLY EMBRYONIC DEVELOPMENT
BEFORE OVIPOSITION
In birds, because the majority of embryonic growth
occurs outside of the maternal body, all of the nutritional
components of the embryo must be included in the egg
at the time of oviposition (lay). As a result of the large
amount of yolk reserves, the active cytoplasm containing
the maternal chromosomes is concentrated on 1 small
area of the yolk called the blastodisc. The blastodisc is
identified as a small whitish area approximately 2 to 3
mm in diameter. If fertilization occurs, during the next
26 h, the blastodisc, while egg formation is occurring
inside the reproductive tract of the hen, undergoes a series
of morphogenetic changes which include cell division
(meiosis and mitosis) and cell differentiation and organi-
zation.
In birds, egg formation and embryonic development
occur as the yolk passes through 5 portions of the oviduct
[infundibulum, magnum, isthmus, shell gland (uterus),
and vagina]. Fertilization takes place in the infundibulum,
and shortly after, the second meiotic division is completed
during the passage of the ovum through the magnum. At
the time the ovum enters the shell gland, approximately 3
to 5 h after fertilization, the first mitotic division occurs
(Patterson, 1910; Olsen, 1942). Cleavage (division) of the
single-celled blastodisc involves numerous mitotic divi-
sions to produce an embryo with thousands of genetically
identical cells. It should be noted that only the cells of
the blastodisc divide and not the yolk.
Shell deposition occurs in the shell gland over the next
18 to 21 h. About 11 h after the development of the first
cleavage furrow, the embryo, which is now called a blas-
toderm, is a circular disc about 5 to 6 cells thick in the
center but tapering out to 1 to 2 cells at the periphery
(Bellairs, 1971). While the ovum is still in the uterus,
the first morphogenetic event in the embryo occurs; this
involves cellular shedding from the central area of the
blastoderm (Eyal-Giladi and Kochav, 1976; stage VII of
embryonic development). The result is that by the time
the egg is laid, the central area of the blastoderm is now
1 to 2 cell layers thick with an area on the periphery that
is several layers thick. The translucent central area of
the blastoderm is called the area pellucida, while the
peripheral “ring” of cells, that is still in contact with the
yolk, is termed the area opaca. It is the ring of cells of
the area opaca that makes up the structure of the embryo
that many industry people call “the doughnut.” The cells
in the area opaca do not contribute to the embryo proper;
only the epiblast cells (that are in the area pellucida)
1021
will eventually form all the structures of the body of
the embryo.
EMBRYONIC ORGANIZATION AT THE
TIME OF OVIPOSITION
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The result of the embryonic developmental processes
that occur while the egg is still being formed in the oviduct
of the hen is that the broiler embryo now consists of
approximately 40,000 to 60,000 cells (Eyal-Giladi and Ko-
chav, 1976; stage X of embryonic development). Although
there may be variations in embryonic development that
occur due to different parent strains and ages, this is the
most common stage of embryonic development at the
time of lay. Gupta and Bakst (1993) have determined that
domestic turkey embryos are at more immature stages
of embryonic development at the time of lay than domes-
tic chickens.
From this point on, unless adequate incubation temper-
atures are provided (37.5°C), the embryo will not continue
normal embryonic development. If the fertile eggs are
provided with proper incubation conditions, and the eggs
are not contaminated to the point where they explode in
the incubator, there are 4 possible outcomes. The embryo
could die during the early (0 to 7 d), middle (8 to 14 d),
or late (15 to 21 d) stages during the 21 d of incubation,
or the embryo could develop and hatch into a chick. One
of the factors that strongly influences this outcome is
egg storage.
EGG STORAGE TEMPERATURE
Both at the breeder farm and at the hatchery fertile
eggs are stored at temperatures below 21°C. One of the
reasons for storing at cool temperatures is to prevent
bacterial growth. The primary reason is to stop the devel-
opment of the embryo.
The temperature below which embryonic development
does not occur has been termed physiological zero. Physi-
ological zero has been reported at 2 different tempera-
tures. Early research by Edwards (1902) found the mini-
mum temperature for embryonic development to be 21°C,
whereas research conducted 40 yr later by Funk and Biel-
lier (1944) reported a higher temperature of 28°C. Myself
and my colleagues (Fasenko et al., 1992) established that
storage of fertile eggs (14°C) for various lengths of time
stopped all embryonic development, as established by
microscopic techniques that differentiate embryonic de-
velopment into stages (Eyal-Giladi and Kochav, 1976).
Using a similar microscopic technique adapted for turkey
embryos (Gupta and Bakst, 1993), Bakst and Gupta (1997)
determined that some turkey embryonic development
during storage for 3, 7, or 14 d at 15°C does occur.
One of the reasons why there may be a discrepancy
between reported minimum temperature requirements
for embryonic development is that the different devel-
oping tissues of the embryo may have varying minimum
temperature requirements for growth to occur and be
readily observable (Kaufman, 1948). Another extremely
1022 FASENKO
important factor that should be further researched is the
concept of physiological zero. Although it may be true
that development using microscopic techniques is not
observable at certain temperatures, there may be cellular
metabolic processes that are still continuing. Because of
this, the term physiological zero seems inappropriate,
and I advocate using the term embryonic diapause as
an alternative.
Although not discussed in depth in the current paper,
there are numerous publications that have examined opti-
mum egg cooler storage temperatures, conditions, and
egg treatments. I recommend seeking out the works of
Proudfoot and Hamilton (1990) and Meijerhof (1992) for
more comprehensive reviews.
THE INFLUENCE OF EGG STORAGE
ON HATCHABILITY
It has been well documented in numerous domestic
fowl that egg storage longer than 1 wk significantly re-
duces hatchability (Scott, 1933; Asmundson, 1947; Kosin,
1950; Becker, 1963; Merritt, 1964; Sittman et al. 1971;
Whitehead et al., 1985; Fasenko et al., 2001a,b). In research
I conducted in which eggs were stored for 2, 4, 6, 8, 10,
12, 14, and 16 d, the results confirmed that storage less
than 8 d does not influence hatchability (Fasenko and
Robinson, 1999). However, hatchability did significantly
decline after storage for 8, 12, and 16 d.
EFFECTS OF EGG STORAGE
ON EMBRYONIC MORTALITY
The negative effect of storage on embryonic mortality
has been noted by researchers for some time (Merritt,
1964; Arora and Kosin, 1966; Sittman et al., 1971; Mather
and Laughlin, 1976). It has been established that embry-
onic mortality even before incubation can be measured
and that mortality increases as storage time lengthens
(Fasenko et al., 1992). In broiler embryos, egg storage for
14 vs. 4 d increased embryonic mortality at early and late
stages of incubation. Overall embryo mortality went from
10.7% in embryos of 4-d stored eggs to 27.7% in embryos
from 14-d stored eggs (Fasenko et al., 2001b). A similar
pattern of embryonic mortality was noted in turkey eggs
stored for 4 (22.9%) vs. 14 d (29.28; Fasenko et al., 2001a).
HOW EGG STORAGE AFFECTS
EMBRYONIC METABOLISM
Research in my laboratory over the past few years has
resulted in the fine-tuning of equipment that is able to
measure the embryonic metabolism of individually incu-
bating eggs over the entire incubation period (Segura et
al., 2006). Using this equipment, it was established that
the metabolism of embryos from 15-d stored eggs, as
measured indirectly by embryonic CO2 output, proceeds
at a slower rate than embryos from 4-d stored eggs (Fa-
senko et al., 2002). This research provides evidence that
embryos from long-term stored eggs not only lag behind
in development, but their metabolism is also changed
due to storage before incubation. Evidence for differences
in embryonic metabolism due to storage is provided by
other research in turkeys (Fasenko, 1996) and broiler
breeders (Christensen et al., 2001). I (Fasenko, 1996)
showed that turkey embryos from 14-d stored eggs de-
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pend more upon gluconeogenesis during pipping and
hatching than do embryos from 4-d stored eggs. Chris-
tensen et al. (2001) examined the carbohydrate metabo-
lism of a line of broiler breeders that was resistant to
long-term storage compared with a broiler breeder line
that was vulnerable to embryonic mortality induced by
storage. They determined that the embryos from the stor-
age-resistant strain were able to maintain larger glycogen
reserves in the muscle and heart than the embryos from
the line more affected by storage. Collectively, the re-
search described above provides evidence that the physi-
ology of an embryo from a long-term stored egg may be
changed in such a way as to be less efficient than an
embryo from a short-term stored egg.
EMBRYONIC DEVELOPMENTAL
CHANGES DUE TO STORAGE
Previous research has established that when eggs are
stored long-term, the incubation time for a chick to hatch
is extended (Kosin and Konishi, 1973; Mather and Laugh-
lin, 1976, 1977). Two possible hypotheses have been put
forth for the increase. Arora and Kosin (1966) showed
that in long-term stored eggs, embryonic development
did not immediately initiate in response to incubation
temperatures. In another study, Mather and Laughlin
(1977) determined that the embryonic development of
embryos from eggs stored for 14 d lagged behind non-
stored eggs by about 12.2 h. In this study, it was estab-
lished that embryonic development in embryos from
stored eggs proceeded at a slower rate during the first
portion of incubation.
Research by myself and my colleagues (Fasenko and
Robinson, 1998) has validated both of these hypotheses.
In this study, it was established that the biological age of
an embryo from a 14-d stored egg lags behind that of an
embryo from a 4-d stored egg (Fasenko and Robinson,
1998). This observation was made even though the chro-
nological ages of the embryos were the same. In examin-
ing embryonic development every 3 hr for the first 12 h
of incubation, it was determined that the development
of the embryos from 14-d stored eggs began to lag behind
as early as 6 hr into incubation. Further to this, it was
determined that not all embryos responded the same way
to long-term storage. Some embryos of long-term stored
eggs, even after exposure to normal incubation tempera-
tures for 12 h, had not initiated any development. Other
embryos advanced in development, but not at the same
rate as embryos from short-term stored eggs. Perhaps the
most interesting result obtained was that there were some
embryos from long-term stored eggs whose development
was equal to that of the short-term stored eggs. If the
factors that provide resistance to long-term storage in
 EMBRYO SYMPOSIUM
these embryos can be identified, the deleterious effects
of long-term storage may be able to be alleviated.
RELATIONSHIP BETWEEN EMBRYONIC
DEVELOPMENT AND THE ABILITY
TO SURVIVE EGG STORAGE
Hays and Nicolaides (1934) conducted one of the first
studies that recognized the relationship between hatch-
ability and embryonic development at the time of lay. In
this research, hens showing a record of consistently
higher hatchability had blastoderms that were at more
advanced stages of development at the time of lay. Taking
advantage of this fact, Kosin, (1956) examined the effect
of preincubation of turkey and chicken eggs before stor-
age. In both species, eggs that were warmed before stor-
age of at least 7 d had higher hatchability than those not
exposed to incubation before storage.
Based on the above findings, further research was con-
ducted in modern turkey and broiler strains by myself
and my colleagues. When turkey breeder eggs were
stored for 4 and 14 d, the hatchability declined from 70.9
to 64.4%, respectively (Fasenko et al., 2001a). However,
when eggs were incubated for 12 h before storage for 14 d,
the embryos had developed to the point where hypoblast
formation was complete, and the hatchability signifi-
cantly improved (70.9%).
In broiler breeder eggs stored for 4 d the hatchability
(89.7%) was significantly lowered when the eggs were
stored for 14 d (72.2%). The negative effect on hatchability
was significantly reduced (78.1%) when the eggs were
incubated for 6 h before storage for 14 d (Fasenko et al.,
2001b). Interestingly, the broiler embryos were at the
same stage of embryonic development (hypoblast forma-
tion complete) after 6 h of incubation as the turkey em-
bryos were after 12 h of incubation.
This study also provided evidence that prestorage incu-
bation does not merely provide extra incubation time for
the embryos to hatch (Fasenko et al., 2001b); when eggs
were incubated for 18 h before storage for 14 d, hatchabil-
ity drastically dropped to 11.5% (Fasenko et al., 2001b).
Collectively, these research results provide evidence that
there are particular embryonic developmental stages that
are better able to survive storage. Embryos that have
completed hypoblast formation may be at a relatively
inactive stage and may withstand developmental arrest
better than embryos that are undergoing active periods
of cellular division, cellular metabolism, differentiation,
or all of these processes.
In all of the above studies conducted by myself and
colleagues, the incubation treatments were imposed be-
fore any egg storage. Because breeder farms are not
equipped with incubators, an additional study by myself
was undertaken to determine if preincubation treatments
after 3 d of on-farm storage would produce the same
positive results on hatchability of long-term stored eggs
(unpublished data). Even though hatchability signifi-
cantly declined in 14- vs. 4-d stored eggs, incubation of
eggs after eggs had been stored for 3 d and before storage
1023
for an additional 11 d did not improve hatchability. The
3 d of storage (simulating on-farm storage) before the
incubation treatments may have somehow reduced the
beneficial effects of the prestorage incubation treatments.
THE NEXT RESEARCH STEP: HOW DOES
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EGG STORAGE AFFECT EMBRYONIC
ABNORMALITIES, CELL NECROSIS,
AND APOPTOSIS?
At the cellular level, Arora and Kosin (1968) observed
that the number of necrotic nuclei in cells of embryos
from stored eggs increased as the length of the storage
period increased. The reduction in healthy cells may be
the reason behind the observations of Mather and Laugh-
lin (1979), who observed that when eggs are stored, blas-
toderm area shrinks.
More recently, Bloom et al. (1998) demonstrated that
there is an increase in the number of cells that are pro-
grammed to die (apoptotic cells) when eggs are stored
for 14 d. Preliminary research conducted in my laboratory
has shown that the percentage of apoptotic cells increases
in embryos of long-term stored eggs vs. embryos from
short-term stored eggs. My hypothesis is that long-term
exposure of embryos to cool storage may increase the
number of necrotic (dead) cells or may predispose the
cells to unscheduled apoptosis. The end result is that the
ratio of nonviable to viable embryonic cells increases and
thus may increase the number of abnormal or dead em-
bryos occurring in long-term stored eggs; an optimum
number of viable embryonic cells is likely required for
initiation of normal growth and development.
ACKNOWLEDGMENTS
I thank the many research colleagues, undergraduate
and graduate student researchers, and research techni-
cians who have made strong contributions to the studies
mentioned in this symposium proceedings. The work
mentioned was supported in part by Alberta Agriculture
Research Institute, Alberta Livestock Industry Develop-
ment Fund, the Agriculture Funding Consortium, Cana-
dian Broiler Hatching Egg Marketing Agency, Lilydale
Hatchery, Maple Leaf Hatchery, Natural Sciences and
Engineering Research Council of Canada, and the Poultry
Industry Council.
REFERENCES
Arora, K. L., and I. L. Kosin. 1966. Developmental responses of
early turkey and chicken embryos to pre-incubation holding
of eggs: Inter- and intra-species differences. Poult. Sci.
45:958–970.
Arora, K. L., and I. L. Kosin. 1968. The response of the early
chicken embryo to pre-incubation temperature as evidenced
from its gross morphology and mitotic pattern. Physiol. Zool.
41:104–112.
Asmundson, V. S. 1947. Time held prior to incubation and hatch-
ability of turkey eggs. Poult. Sci. 26:305–307.
1024 FASENKO
Bakst, M. R., and S. K. Gupta. 1997. Preincubation storage of
turkey eggs: Impact on rate of early embryonic development.
Br. Poult. Sci. 38:374–377.
Becker, W. A. 1963. Length of preincubation storage of turkey
eggs and its effects on body weight. Poult. Sci. 42:1356–1359.
Bellairs, R. 1971. Order and adjustment in the early stages of
bird development. Page 69 in Developmental Processes in
Higher Vertebrates. Logos Press Ltd., London, UK.
Bloom, S. E., D. E. Muscarella, M. Y. Lee, and M. Rachlinski.
1998. Cell death in the avian blastoderm: Resistance to stress
induced apoptosis and expression of anti-apoptotic genes.
Cell Death Differ. 5:529–538.
Christensen, V. L., M. J. Wineland, G. M. Fasenko, and W. E.
Donaldson. 2001. Egg storage effects on plasma glucose and
supply and demand tissue glycogen concentrations of broiler
embryos. Poult. Sci. 80:1729–1735.
Edwards, C. L. 1902. The physiological zero and the index of
development from the egg of the domestic fowl. Am. J. Phys-
iol. 6:351–397.
Eyal-Giladi, H., and S. Kochav. 1976. From cleavage to primitive
streak formation: A complementary normal table and a new
look at first stages of the development of the chick. I. General
morphology. Dev. Biol. 49:321–337.
Fasenko, G. M. 1996. Factors influencing embryo and poult
viability and growth during long term storage of turkey eggs.
PhD Thesis. North Carolina State Univ., Raleigh.
Fasenko, G. M., V. L. Christensen, M. J. Wineland, and J. N.
Petitte. 2001a. Examining the effects of prestorage incubation
of turkey breeder eggs on embryonic development and
hatchability of eggs stored for four or fourteen days. Poult.
Sci. 80:132–138.
Fasenko, G. M., and F. E. Robinson. 1998. Identification of the
incubation period when broiler breeder embryonic develop-
ment is delayed due to egg storage for 14 versus 4 days.
Poult. Sci. 77(Suppl. 1):77.(Abstr.)
Fasenko, G. M., and F. E. Robinson. 1999. Profiling egg storage:
The effects on egg weight loss, egg characteristics, and hatch-
ability. Poult. Sci. 78(Suppl. 1):9.(Abstr.)
Fasenko, G. M., F. E. Robinson, R. T. Hardin, and J. L. Wilson.
1992. Variability in preincubation embryonic development
in domestic fowl. 2. Effects of duration of egg storage period.
Poult. Sci. 71:2129–2132.
Fasenko, G. M., F. E. Robinson, J. C. Segura, J. J. R. Feddes, and
C. A. Ouellette. 2002. Long term hatching egg storage alters
the metabolism of broiler embryos. Poult. Sci. 80(Suppl. 1):62.
Fasenko, G. M., F. E. Robinson, A. I. Whelan, K. M. Kremeniuk,
and J. A. Walker. 2001b. Prestorage incubation of long-term
stored broiler breeder eggs. 1. Effects on hatchability. Poult.
Sci. 80:1406–1411.
Funk, E. M., and H. V. Biellier. 1944. The minimum temperature
for embryonic development in the domestic fowl (Gallus do-
mesticus). Poult. Sci. 23:538–540.
Gupta, S. K., and M. R. Bakst. 1993. Turkey embryo staging
from cleavage through hypoblast formation. J. Morphol.
217:313–325.
Hays, F. A., and C. N. Nicolaides. 1934. Variability in develop-
ment of fresh laid hens eggs. Poult. Sci. 13:74–89.
Kaufman, L. 1948. The effect of certain thermic factors on the
morphogenesis of fowl embryos. Pages 351–356 in Official
Downloaded from https://academic.oup.com/ps/article-abstract/86/5/1020/1620704 by Universidade Federal do Rio Grande do Sul user on 15 September 2018
Report of the Eighth Worlds’ Poultry Congress. J. Baelum and
A. Nielsen, ed. Nielsen and Lydich, Copenhagen, Denmark.
Kosin, I. L. 1956. Studies on pre-incubation warming of chicken
and turkey eggs. Poult. Sci. 35:1384–1392.
Kosin, I. L. 1950. A relationship between the length of storage
and incubation periods in Broad Breasted Bronze eggs. Poult.
Sci. 29:620–621.
Kosin, I. L., and T. Konishi. 1973. Pre-incubation storage condi-
tions and their effect on the subsequent livability of chicken
embryos: Exogenous CO2, plastic bags and extended holding
periods as factors. Poult. Sci. 52:296–302.
Mather, C. M., and K. F. Laughlin. 1976. Storage of hatching
eggs: The effect on total incubation period. Br. Poult. Sci.
17:471–479.
Mather, C. M., and K. F. Laughlin. 1977. Storage of hatching
eggs: The effect on early embryonic development. Br. Poult.
Sci. 18:597–603.
Mather, C. M., and K. F. Laughlin. 1979. Storage of hatching
eggs: The interaction between parental age and early embry-
onic development. Br. Poult. Sci. 20:595–604.
Meijerhof, R. 1992. Pre-incubation holding of hatching eggs.
Worlds’. Poult. Sci. J. 48:57–68.
Merritt, E. S. 1964. Pre-incubation storage effects on subsequent
performance of chickens. Br. Poult. Sci. 5:67–73.
Olsen, M. W. 1942. Maturation, fertilization, and early cleavage
in the hen’s egg. J. Morphol. 70:513–533.
Patterson, J. T. 1910. Studies on the early development of the
hen’s egg. I. History of the early cleavage and of the accessory
cleavage. J. Morphol. 21:101–134.
Proudfoot, F. G., and R. M. G. Hamilton. 1990. Care of hatching
eggs before incubation. Publication 1573/E. Commun.
Branch, Agric. Canada, Ottawa, Ontario.
Scott, H. M. 1933. The effect of age and holding temperature on
hatchability of turkey and chicken eggs. Poult. Sci. 12:49–54.
Segura, J. C., C. Ouellette, J. J. R. Feddes, G. M. Fasenko, and
M. J. Zuidhof. 2006. Development of a metabolic calorimeter
system to measure heat production of domestic avian em-
bryos during incubation. Can. Bios. Engin. 48:41–46.
Sittman, K., H. Abplanalp, and C. F. Myerdick. 1971. Extended
storage of quail, chicken and turkey eggs. 1. Hatchability
and embryonic mortality. Poult. Sci. 50:681–688.
Whitehead, C. C., M. H. Maxwell, R. A. Pearson, and K. M.
Herron. 1985. Influence of egg storage on hatchability, em-
bryonic development, and vitamin status in hatching broiler
chicks. Br. Poult. Sci. 26:221–228.