Influence of Oxygen Concentration on Hatchability
and on Selecting for Hatchability1,2'3
G E O R G E T.
University of Wyoming, Laramie, Wyoming
(Received for publication April 21, 1954)
O B T A I N I N G a respectable hatching
percentage from the eggs of domes-
tic poultry is one of the important prob-
lems of the poultry industry in areas of
high elevation. This problem exists a t
Laramie, Wyoming, where the elevation
is approximately 7,200 feet above sea
level. T h e Wyoming Agricultural Experi-
ment Station, highest in the United
States, is an ideal location to study the
problems of high-altitude incubation.
One objective of the investigation re-
ported in this paper was to reexamine the
reported improvement in hatchability re-
sulting from the use of supplemental oxy-
gen in incubation. Another objective was
to study the effect of oxygen concentra-
tion as it is related to selecting for genetic
improvement in hatchability.
North (1941) stated t h a t the lower
barometric pressure of Laramie lowers the
volumetric concentration of oxygen b y 25
percent and t h a t more ventilation is re-
quired to supply the same amount of oxy-
gen in the incubator. Thompson (1952)
reported t h a t at altitudes of 6,000 feet and
higher, most incubators decline in effici-
ency and t h a t above 8,000 feet they be-
1
Published with approval of the Director,
Wyoming Agricultural Experiment Station, as
Journal Paper No. 49.
2
The oxygen used in this experiment was fur-
nished by the Medical Gas Division of the Denver
Oxygen Company, Denver, Colorado, Harold Rogers,
Manager.
3
This paper is based on a thesis submitted by
the author to the faculty of the Graduate School,
University of Wyoming, in partial fulfillment of the
requirements for the degree of Doctor of Philosophy.
107
DAVIS
come virtually useless. Wilgus (1949) ob-
served the physical factors within incu-
bators used at Fort Collins, Colorado, a t
an elevation of 5,100 feet. He found air
speed to vary as much as 10 times and
temperature to vary as much as 2 | de-
grees Fahrenheit in different locations
within the same incubator. He pointed
out the desirability of having oxygen and
carbon dioxide indicators on the incu-
bators to furnish a sound basis for ventila-
tion control.
A study of the effect of oxygen concen-
tration on the development of the chick
embryo was made b y Wesselkin (1913).
Growth and development were retarded
in eggs incubated in atmospheres contain-
ing 5, 10, and 15 percent oxygen. T h e de-
gree of retardation was associated nega-
tively with oxygen concentration. Barott
(1937) reported t h a t when eggs were in-
cubated in atmospheres containing 15, 18,
21, 30, 40, and 50 percent oxygen, the best
hatch was obtained at 21 percent. Below
21 percent, the decrease in hatchability
was 5 percent for each 1 percent decrease
in concentration. Riddle (1924) concluded
t h a t most bird embryos require an atmos-
phere of no less than 15 percent oxygen
at some stage of development.
Arbuckle (1918) found t h a t oxygen in
the incubator could be depleted b y re-
stricting ventilation in the late stages of
incubation. This depletion resulted in
lower hatchability.
Ells and Morris (1947) and Meshew
(1949) investigated the use of supple-
mental oxygen in incubation a t Laramie,
108 G. T. DAVIS
Wyoming. They found that the hatcha-
bihty of chicken and turkey eggs could be
increased by raising the oxygen concen-
tration to 25 percent. Volumetrically this
concentration is approximately the same
as 19 percent oxygen at sea level. Stephen-
son (1950a, b) reported similar results
obtained at Logan, Utah, at an elevation
of 4,500 feet. His work indicated that the
improvement in hatchability was propor-
tional to the length of time supplemental
oxygen was used.
North (1944) reported that strains of
chickens obtained from low-altitude loca-
tions where eggs have been hatching satis-
factorily will seldom produce good hatches
at high altitude. After several generations
at high altitude, such strains hatch nearly
as well as strains at low altitude.
Shoffner and Sloan (1948) estimated the
heritability of hatchability of chicken eggs
to be 16 percent. They employed the
method of dam-daughter regression with-
in sire groups to make their estimate.
The regression did not differ significantly
from zero. Wilson and Johnson (1946)
also used dam-daughter regression to esti-
mate the heritability of hatchability of
turkey eggs. Their estimate was 26 per-
cent. Abplanalp and Kosin (1953) used
several methods in estimating heritability
of hatchability of turkey eggs. Their esti-
mates varied from 3 percent to 64 percent.
They reported that eggs held 8 to 14 days
before setting gave higher estimates than
those held 1 to 7 days before setting.
Hammond (1947) expressed the hypo-
thesis that characteristics can be best
selected for under environmental condi-
tions which favor their fullest develop-
ment. Thus optimal environmental condi-
tions would be essential to improvement
by selection. Falconer and Latyszewski
(1952) selected for growth rate in mice
under two environmental conditions.' One
line was fed a full ration and another a
restricted diet. Selection for growth was
effective in both lines but heritability was
higher in the line which received the re-
stricted diet. When the full-diet line was
reared on a restricted diet, it was inferior
to the restricted-diet line and showed no
improvement over an unselected re-
stricted-diet line. When the restricted-
diet line was reared on a full diet, it was
nearly as good as the full-diet line and
showed marked improvement over an un-
selected full-diet line. Thus optimal en-
vironmental conditions were effective for
selecting only when optimal environment
would be continued, but sub-optimal en-
vironment was effective for selecting for
growth when either environmental condi-
tion would prevail.
MATERIALS AND METHODS
The experiment was designed to com-
pare the hatching records of hens when
their eggs were incubated under two en-
vironments. These were normal atmos-
pheric air, which will hereafter be referred
to as air, and normal atmospheric air sup-
plemented with oxygen, which will be
called oxygen in subsequent descriptions
and discussions. The amount of oxygen
used was that quantity necessary to raise
the oxygen level within the incubators to
approximately 25 percent. Volumetrically,
this is nearly equal to an atmosphere con-
taining 19 percent oxygen at sea-level
pressure. In all hatches the eggs from each
hen were divided equally and set in both
environments.
New Hamsphire chickens were used in
the study. The study was started in the
1949-50 hatching season and continued
for three seasons.
In the fall of 1949, 210 pullets were
flock-mated to 15 cockerels. From this
flock 96 pullets were selected for the first
trial. These were selected for high rate of
 INFLUENCE OF OXYGEN ON HATCHABILITY
egg production in order to insure a suffi-
cient number of eggs from each hen. They
were trap-nested and the eggs were marked
for pedigree hatching.
The 1950-51 trial was conducted with
the offspring of the birds used in 1949-50.
These offspring were divided into two
groups, one of which had been hatched in
oxygen and the other in air. Each pen in-
cluded 260 pullets and 15 cockerels. From
each pen 60 pullets were selected for the
trial. They were selected so that as many
dams as possible were represented in each
group. Fifty-three dams were represented
in the group hatched in oxygen and 38
dams in the group hatched in air. Cock-
erels were hatched in same environment
as the pullets. Again the pullets were trap-
nested and the eggs marked for pedigree
hatching.
In the 1951-52 trial, birds used were the
offspring of those pullets used in the 1950-
51 trial. The pullets were selected so that
each dam represented in the trial would
have no less than two nor more than four
daughters hatched in each of the incu-
bator atmospheres. The pullets which
were hatched in oxygen were housed to-
gether, and those hatched in air were
housed together. Thus each flock included
pullets hatched in one atmosphere, but
each included some pullets whose dams
were hatched in oxygen and some whose
dams were hatched in air. Males mated to
oxygen hatched pullets were themselves
hatched in oxygen; their dams likewise
were hatched in oxygen. Males mated to
pullets hatched in air were themselves
hatched in air; their dams likewise were
hatched in air. There were 260 pullets and
17 cockerels in each flock. Seventy-nine
pullets were selected from the flock
hatched in oxygen and 80 were selected
from the flock hatched in air. These were
offspring of 24 dams, each of which had
daughters in both flocks. The pullets were
109
trap-nested and the eggs were marked for
pedigree hatching.
In the first and second years there were
six hatches and in the third year there
were three hatches. Three forced-draft
incubators of similar construction, each
with a capacity of 384 eggs, were em-
ployed in the project. In order to mini-
mize the individual effects of each incuba-
tor, each environment was employed in
each incubator an equal number of times.
Oxygen was introduced into the oxygen
incubator through a gas-flow regulator.
Ventilators were adjusted to keep the oxy-
gen level constant. Hatching records were
kept for each hen under each hatching en-
vironment and the chicks pedigree identi-
fied.
RESULTS
Factors Causing Failure to Hatch. The
1949-50 hatches were examined to de-
termine the percentage of infertile eggs
and the stages of incubation when embry-
onic mortality occurred. The eggs were
candled on the fifth day of incubation to
determine fertility and on the eighteenth
day to determine embryonic mortality
which had occurred up to that time. Those
which contained live embryos on the
eighteenth day but which failed to hatqh
were examined after the chicks were re-
moved on the twenty-second day of incu-
bation to determine whether the em-
bryos died before pipping. The incidence
of infertility and mortality at various
stages of incubation was lower in eggs in-
cubated in oxygen as compared with those
incubated in air. A summary of infertility
and stages when mortality occurred is pre-
sented in Table 1.
Because of the low incidence and ap-
parent inaccuracies in its determination,
infertility is not here regarded as a separ-
ate factor from embryonic mortality. The
term hatchability as used hereafter will
 110 G. T. DAVIS

TABLE 1.—A comparison of infertility and stages of mortality of eggs set

with and without supplemental oxygen, 1949-50

Infertile 18 day 18-21 day Pipped

Number mortality mortality eggs
Incubation
of eggs
treatment set Num- Per- Num- Per- Num- Per- Num- Per-
ber cent ber cent ber cent ber cent

Oxygen 1,149 36 3.1 238 20.7 130 11.3 45 3.9

Air 1,149 52 4.5 340 29.6 252 21.9 71 6.9
Differences 0 16 1.4 116 8.9 122 10.6 26 3.0

be the percentage of all eggs set which
hatch rather than the percentage of fer-
tile eggs which hatch.
Sources of Variation in Hatchability.
Hatching records for the 1949-50 season
were analyzed to determine the causes of
variation in hatching percentages. Analy-
sis of covariance as described by Snedecor
(1946) was employed in the analysis. The
number of eggs set was used as the inde-
pendent variable and the number of chicks
hatched was the dependent variable. The
variations tested were those caused by
differences in individual hens, time of set,
oxygen environment in the incubator,
and the interactions between each of these
three causes of variation.
Differences at the .01 level of proba-
bility were found among individual hens,
times of set, oxygen environment, and in-
teractions between hens and sets, and be-
tween sets and oxygen environment. The
interaction between hens and oxygen en-
vironment was non-significant. This may
be interpreted as meaning that the pheno-
type for hatchability of the hen may be
measured by the record she establishes in
either oxygen or air incubation, or when
these records established under both en-
vironments are combined. Her records
established in oxygen or air may be con-
sidered as valid measurements of her
hatching ability.
The Effect of Supplemental Oxygen on
Hatching. Hatchability of each of the 15
hatches in the three-year period of the ex-
periment was improved by supplemental
oxygen. In the 1949-50 season, the im-
provement per 100 eggs set was 22.2
chicks, for 1950-51, 11.7 chicks, and for
1951-52, 12.4 chicks.
The data disclose an annual improve-
ment in eggs hatched in air. Hatchability
for the eggs was, for successive years, 38.1,
44.5 and 49.9 (Table 2). The regression of
improvement per year was found to be
5.51 + 1.05 percent, which is highly signif-
icant. For eggs set in oxygen, percentages
for corresponding hatches were 60.3,
56.2, and 62.3, which provided an annual
improvement of .0079 + 3.10 percent,
which is non-significant.
Hatching records of the 39 pullets which
were hatched in oxygen, and whose dams
were hatched in oxygen, were compared
with those of the 37 pullets which were
hatched in air, and whose dams were
hatched in air. Hatchability records es-
tablished in air for the two groups were
respectively 43.5 and 50.9 percent. These
percentages were just short of being sig-
nificantly different at the .05 level. Eggs
set from these groups in oxygen resulted in
hatches of 57.3 and 63.5 percent respec-
tively. These differences are non-signifi-
cant; however, when the records for both
environments are combined, the resulting
percentages (50.2 and 57.2) are signifi-
cantly different at the .05 level.
This improvement caused by the use of
 INFLUENCE OF OXYGEN ON HATCHABILITY 111

TABLE 2.—Summary of hatching for 15 sets of eggs

Hatched with supplemental Hatched in unsupplemented

Increased
oxygen air number of
Set chicks
Date set Number Number Number Number
number Percent Hatched per 100
eggs chicks eggs chicks
hatched set percent eggs set
set hatched hatched

1 Nov. 7, 1949 151 104 69.8 151 65 43.0 26.8

2 Dec. 5, 1949 193 138 71.5 193 68 35.2 36.3
3 Jan. 2, 1950 153 92 60.1 153 50 32.7 27.4
4 Jan. 30, 1950 207 137 56.2 207 86 41.5 24.7
5 Feb. 27, 1950 194 75 38.7 194 70 36.1 2.6
6 Mar. 27, 1950 186 108 58.1 186 74 39.8 18.3
Totals for 1949-50 1,084 654 60.3 1,084 413 38.1 22.2
7 Nov. 27, 1950 314 158 50.3 314 119 37.9 12.4
8 Dec. 25, 1950 349 175 50.1 349 152 43.6 6.5
9 Jan. 22, 1951 308 174 56.5 308 139 45.1 11.4
10 Feb. 19, 1951 278 181 65.1 278 141 50.7 14.4
11 Mar. 19, 1951 283 167 59.0 283 130 45.9 13.1
12 Apr. 16, 1951 258 151 58.5 258 115 44.6 13.9
Totals for 1950-51 1,790 1,006 56.2 1,790 796 44.5 11.7
13 Jan. 30, 1952 345 205 57.9 354 168 47.5 10.4
14 Feb. 27, 1952 336 234 69.6 336 172 51.2 18.4
15 Mar. 26, 1952 303 180 59.4 303 156 51.5 7.9
Totals for 1951-52 993 619 62.3 993 496 49.9 12.4
Totals for 3 years 3,867 2,279 58.9 3,867 1,705 44.1 14.8

oxygen was nearly equal for both groups.
The actual increases in number of chicks
per 100 eggs set were 13.8 and 12.6 re-
spectively. When computation was made
to determine the gain as a percentage of
maximum possible improvement (gain in
chicks per 100 eggs divided by 100 minus
hatching percentage) both groups re-
sponded almost exactly alike. Computa-
tions are as follows:
13.8
For oxygen -=.2404
100-43.5
12.6
For air -=.2570
100-50.9
These results indicate not only that
hatchability improves more rapidly each
generation when oxygen is not used in the
incubator, but also that the improvement
does not lower the eggs' capacity to be
further improved by being set in oxygen.
This may be interpreted to mean that the
gain resulting from successive years'
hatching in air is a genetic improvement
that is reflected in either of the incubator
environments and it is not merely a gene-
tic resistance to low oxygen concentra-
tion.
Factors Involved in Hatching Improve-
ment. Because the analysis of covariance
indicated that records of hatchability es-
tablished in oxygen are equivalent to such
records established in air, the records es-
tablished under each environment for
each hen were combined for further an-
alysis. Combining these data allowed the
use of larger samples of hatching ability
for each hen without increasing the experi-
mental error.
To further investigate the possibility
that the records established in both en-
vironments are related, a partial correla-
tion was determined between the number
of chicks hatched under each environ-
ment, independent of the number of eggs
set. The resulting correlation coefficient
is .7897, which is highly significant at the
.01 level.
A comparison of grandams' hatching
112 G. T. DAVIS
records was made between the line de-
veloped in successive hatching in oxygen
and the line developed in successive hatch-
ing in air. The hatching percentage of
those producing the oxygen line was 58.26
and of those producing the line developed
in air, 74.40. The 1949-50 flock, which
was the grandam generation, was a single
flock, and both lines were developed from
it. Considerably more selection pressure
was exerted on the line developed in air
incubation. This was natural selection
since every dam which had at least one
daughter at the time the next year's trial
began was represented in the second gen-
eration. Only 38 of the 96 hens had daugh-
ters which were hatched in air while 53
had daughters which were hatched in
oxygen.
Since the pullets for the last year's trial
were selected to provide a minimum of
two daughters hatched in each environ-
ment, some selection pressure was applied
to both lines. Of the 24 families selected,
11 were from the line hatched in air and
13 were from the oxygen line. Thus there
were nearly as many superior families re-
sulting from the 37 air-hatched hens of
the first trial as there were from the 52
hens of the oxygen-hatched hens.
Heritability estimates were computed
for the oxygen-hatched line and the air-
hatched line. The method of doubling the
regression of dam on daughter was em-
ployed. The regessions were computed
within subclasses in order to eliminate re-
gression caused by differences in years.
The resulting regressions and heritability
estimates are as follows:
6 B*
Oxygen line .146±.137 .292
Airline .324 ±.158 .648
The two regression coefficients are signifi-
cantly different at the .05 level.
In order to find some explanation for
the differences in heritability, coefficients
of variability were computed for the
chicks hatched in 1949-50. For the chicks
hatched in oxygen this coefficient was
.6247 and for those hatched in air, .5344.
Evidently heritability was higher in the
line hatched in air because variation
caused by environment was smaller.
DISCUSSION
Evidence presented by investigators
Ells and Morris (1948), Meshew (1949),
and Stephenson (1950a, b) and the data
obtained in this experiment leave little
doubt that hatchability can be improved
by use of supplemental oxygen during in-
cubation at high altitudes. Hatcherymen
could well take advantage of this informa-
tion provided the gain in hatchability
more than offsets the cost of installing and
maintaining oxygen apparatus.
Hatcherymen, however, should not
overlook the possibility that they may
make genetic improvement in hatchability
in their flocks by not supplementing oxy-
gen. Such incubation could be limited to
hatching of chicks which will be used in
future breeding flocks.
The experiment indicates the possibility
that high-altitude locations may furnish
environment for more rapid improvement
than low-altitude locations provide. It
does not indicate what altitude is optimal
for such selection. The possibility exists
that an altitude higher than that of Lara-
mie may be more desirable. Perhaps high
altitude may be artificially created at low-
altitude location by diluting the air with-
in the incubator with an inert gas such as
nitrogen.
The question of what level of environ-
ment is best for most effective selection
has not been satisfactorily answered.
Hammond (1947) suggests an optimal
one, Falconer and Latyszewski (1952)
suggest one in which the offspring are to
be perpetuated, and Abplanalp and Kosin
 INFLUENCE OF OXYGEN ON HATCHABILITY
(1953) indicate more rapid progress with
a sub-optimal one. This investigation sug-
gests that sub-optimal environment may
be superior for selecting, provided the
character being selected responds in like
degree to varying levels of environment,
and provided variation caused by environ-
ment is reduced.
SUMMARY
A study was made on the effect of sup-
plemental oxygen on hatchability and on
selecting for hatchability in New Hamp-
shire chicken eggs at high altitude (7,200
feet).
From a common foundation stock two
lines were established, one successively
hatched in an incubator in which the air
was supplemented with oxygen, and the
other hatched without supplemental oxy-
gen. Hatchability records for each hen
were established in both environments
throughout the study.
In all hatches a higher percentage of
hatchability was obtained in the incuba-
tion with oxygen supplementation.
The hens were found to respond in like
degree in hatchability to both environ-
ments. The oxygen levels provided equi-
valent environments for genetic expres-
sion of hatchability.
Incubation without oxygen supple-
mentation resulted in genetic improve-
ment in hatchability, while no such im-
provement occurred in the line hatched
successively in supplemented air.
This improvement was found to be due
to natural selection and to higher herita-
bility of hatchability in the unsupple-
mented line. The higher heritability was
apparently caused by reduction in en-
vironmental variability.
REFERENCES
Abplanalp, H. M., and I. L. Kosin, 1953. Genetic
variation of fertility and hatchability in the
113
Broad Breasted Bronze turkey. Poultry Sci. 32:
321-331.
Arbuckle, H. B., 1918. Report of an investigation as
to cause of death of chicks in shell in artificial
incubation. J. Elisha Mitchell Soc. 34: 141-145.
Barott, H. G., 1937. Effect of temperature, humidity,
and other factors on hatch of hens' eggs and on
energy metabolism of chick embryos. U.S.D.A.
Tech. Bull. 553.
Ells, J. B., and L. Morris, 1947. Factors involved in
hatching chicken and turkey eggs at high eleva-
tions. Poultry Sci. 26: 635-638.
Falconer, D. S., and M. Latyszewski. 1952. The
environment in relation to selection for size in
mice. J. Genetics, 51: 67-80.
Hammond, J., 1947. Animal breeding in relation to
nutritional and environmental conditions. Biol.
Rev. 22: 195-213.
Meshew, M. H., 1949. The use of oxygen in the
hatching of chicken and turkey eggs at high alti-
tudes. Poultry Sci. 28: 87-97.
North, M. O., 1941. High altitude incubation of eggs.
U. S. Egg and Poultry Mag. 47:158-159, 184.
North, M. O., 1944. High altitude incubation of
chicken and turkey eggs. Wyoming Agr. Exp.
Sta. Pamphlet 10.
Riddle, O., 1924. On the necessary gaseous environ-
ment of the bird embryo. Ecology, 5:348-362.
Shoffner, R. N., and H. J. Sloan, 1948. Heritability
studies in the domestic fowl. Report on the
Eighth World's Poultry Congress, Copenhagen,
1:269-281.
Snedecor, G. W., 1946. Statistical Methods. The
Iowa State College Press, Ames, Iowa, 1-485.
Stephenson, A. B., 1950a. Supplemental oxygen for
high altitude incubation. Poultry Sci. 29: 781.
Stephenson, A. B., 1950b. Supplemental oxygen in-
creases hatchability of eggs at high altitudes.
Utah Agr. Exp. Sta. Farm and Home Sci. 11: 65,
95-96.
Thompson, R. L., 1952. Incubation at high altitudes.
The effects of wind, barometric pressure and
humidity on foetal mortality in the hen's eggs.
Poultry Sci. 31: 497-509.
Wesselkin, N., 1913. Ueber den Einfluss' des Sauer-
stoffmangels auf des Wachstum und die Ent-
wicklung von Huhnerembryonen. Zentralblatt
fur Allgemeine Pathologie und Pathologische
Anatomie, 24: 1033-1034.
Wilgus, H. S., 1949. High altitude studies show need
for better incubator controls. U. S. Egg and
Poultry Mag. 55 (9): 15.
Wilson, W. O., and L. E. Johnson, 1946. The inher-
itance of egg production and hatchability in
turkeys. Poultry Sci. 25: 278-284.