RESPIRATORY MOVEMENTS OF THE CHICKEN DURING

THE PARAFOETAL PERIOD

by
c. ROMIJN
(Laboratory of Veterinary Physiology, Utrecht)
Received June 27, I947
INTRODUCTION.
From the work of ROMIJN and Roos (1938) we know that the air space of the hen's
egg plays an important part in foetal development. Especially during the last day of
incubation when the chicken has perforated the membranous base of the air chamber and
lungventilation has begun, the animal respires from an atmosphere with an extremely
high carbon dioxide percentage, (up to 9.11 %) and a very low oxygen percentage (9%
or less). During this S.c. parafoetal period the respiration by means of the allantoic vessels
is gradually superseded by real lung ventilation.
WINDLE and BARCROFT (1938) state that respiratory movements were never seen
before the 17th or 18th day (sometimes not before the 19th), although movements of
the muscles involved in respiration appeared very much earlier. From the observations
of these authors on the effect of various amounts of carbon dioxide and oxygen on the
general movements of chickens during the second half of the incubation time, we may
conclude that the parafoetal period must be of great physiological importance for the
initiation of a respiratory rythm of
the animal. The author studied the
F
development of the respiratory mo
vements during the last day of in
cubation and the results have been
collected in the present communi
cation.
METHODS.
A series of 50 eggs of White
Leghorn hens were incubated at a
temperature of 40 C. At the end of
the 19th day of incubation some
eggs (E) were isolated in a small
incubator at the same temperature
and a small funnel was fixed on
the blunt end of it, after artificial
T
L2 0
~
c........==
Inc
L,
s
perforation of the shell on that
spot. The funnel was connected by
Fig. 1. Optical registration of fluctuations in pressure in
means of narrow, flexible rubber
the air-chamber of the incubated egg.
tube to a glass funnel (R), closen
with a very thin rubber membrane
of about 0.01 mm thickness and a diameter of about 6 mm. Bij this method the air chamber
of the incubated egg is connected with the air in the glass funnel and fluctuations in
From: Physiologica Comparata et Oecologia 1(1): 24-28 (1948)
---------
'----;-1----
A
1 1 ~ . ~ ' 1 ~ ,., \ . ~ I ' I I 1 ' ~ ' (.' II' ,,'I,' .'
, I ~ , ~ ~ I
.-"K!----- ;z /
B C
.-.
D
------------ ------
( /'
E F
Fig. 2. Development of respiratory movements in the hen's embryo.
25 C. ROMIJN: PARAFOETAL RESPIRATORY MOVEME:-ITS OF THE CHICKEN
pressure within the air chamber could be recorded optically. A silvercovered piece of
coverglass, was fitted on a small segment of the rubber membrane and a beam of light
from lamp A, thrown on the mirror reflected by it on the slit of a camera with a cylindrical
lens (C) and a film of photographic paper (F). A second beam of light from lamp B was
projected on the same film and was interrupted at regular intervals for time-recording
purposes.
RESULTS.
From the end of the 19th day till the end of the 20th day of incubation when the
chicken perforates the egg shell which covers the air chamber, the movements of the embryo
could be recorded. Some results have been collected in fig. 2. During the 19th day of
incubation embryonic movements are very rare and only one deflection of the beam of
light could be recorded every 10 minutes.
Starting with the 20th day of incubation (A) there is an increase in rythm, though
the movements are still irregular in intensity and frequency. Probably at this moment
the membranous base of the air chamber is perforated by the beak of the animal and
it respires directly from the air in the air space, together with the respiration by means
of the allantoic circulation which is still in full development. At this moment of foetal
development lungventilation begins, a situation which is characteristic of the bird's
embryo and not comparable with any stage of development in the respiration of mammal
foetuses.
A few hours after the beginning of these movements the rythm is more regular and
the fluctuations in pressure are of the characteristically respiratory type (B). Periods
of great frequency are followed by periods of slower rythm. The great sensitiveness of
the recording technique could be demonstrated by increasing the speed of the paper
film (D). The great fluctuations of the zero line coincide with the respiratory movements,
the smaller with the heartbeat of the embryo. At this stage of development (19 days and
10 hours) respiratory movements have a frequency of about 15 a minute, the heart beats
with a rythm of about 200 in the same space of time. During the following hours the
respiratory rythm increases steadily and for some hours the frequency remains relatively
constant (C). The respiratory movements are interrupted at irregular intervals by
movements of the body of the embryo (t).
At the end of the 20th day of incubation, the frequency becomes faster and faster (E).
A rythm of 60 respiratory movements per minute and more is not very rare. General
mass movements of the embryo are more regular and appear at short intervals and during
the last hour of the 20th day or at the beginning of the 21st day the movements of the
body are very energetic and frequent. At this stage of development the animal tries to
break the shell and when succeeding, the fluctuations in pressure stop (F). During the
last minutes of the parafoetal period, when the movements of the body predominate,
the respiratory rythm is of such high frequency, that an accurate recording was not very
well practicable. With six other eggs identical results were obtained; the results obtained
with four of them have been collected in the following table (table J).
26 C. ROMIJN: PARA FOETAL RESPIRATORY MOVEMENTS OF THE CHICKEN
Table 1. Development of respiratory movements in the chicken.
I Frequency of respi
Egg(Nr) Incubation time ) ratory movements Remarks
(per minute)
I!J days 23 hours I 65
20 75
20 2 min. 80
20 15 94
20 40 120
20 55 150
20 62 150
20 100" 150
20 2 hours 120
20 2! " 130
20 3 135
2
3
4
20
20
20
20
20
19
19
19
19
19
19
19
19
19
19
20
20
20
19
19
19
19
19
19
19
19
19
i 19
3hours
II!hours

13
14
14! "
16
In "
20
21
23
I
I! "
7 hours
8
"
9
"
10
12
13
13!
15
15
II
52
69
2-3
9-10
40
25
30
25
20
35
50
60
64
80
II
15
13-14
50
so
80-24
70 -30
80--30
70-30
the animal squeaks within the shell
shell still intact
the animal squeaks
CO
2
content of the air in the chamber 7%
egg shell perforated
the animal squeaks (lungventilation from the air
chamber)
shell perforated
general movements of the foetus
irregular foetal movements
irregular body movements of low frequency.
regular body movements
movements of the respiratory type, regular; the
animal squeaks for the first time (Iungventil
ation I)
respiratory movements very regular, every 10 min.
interrupted by squeaking movements
. increasing muscular activity of the chicken
CO
2
content of the air chamber gas 8.43%
the animal perforates the shell
regular movements of the respiratory type, the
membranous base of the air chamber still intact
animal respires from the air in the air chamber,
lungventilation
regular respiratory movements, interrupted by
squeaking
periods 0 f high frequency, followed by low activity
incl'easing intenSIty of body movements
perforating of the shell

From the data obtained, some conclusions of general importance in relation to the
development of respiration can be drawn. First of all, respjratory movements exist in the
chick before lungventilation is established (eggs 3 and 4), though they are not of the
regular type. In the White Leghorn chicken, incubated at a temperature of 40 C they
27 C. ROMIJN: PARAFOETAL RESPIRATORY MOVEMENTS OF THE CHICKEN
can be demonstrated during the first half of the 20th day of incubation and even earlier.
From the figures collected in our first communication (1938) on the composition of the
gas in the air chamber (l.c. p. 374) we know that at the end of the 19th day of incubation
there is a sudden increase in CO
2
percentage and decrease in O
2
percentage. At this stage
of development the CO
2
content takes up about 5% and the oxygen content about 15%.
As the total gas pressure in the air chamber will be about atmospheric, the blood in the
allantoic vessels is balanced with a gas mixture comparable with the alveolar air of
mammals.
Starting with the end of the 19th breeding day the composition of the gas alters, as a
result of the increasing metabolism of the foetus. The initiation of regular respiratory
movements during the first hours of the last two breeding days is in accordance with
these findings. According to \\TINDLE and BARCROFT (1938) all movements are stimulated
by the carbon dioxide.
Perforation of the membranous base of the air chamber by the beak of the animal
should be considered as a result of the increasing activity of the neck muscles. At this
moment lungventilation begins and the embryo respires directly from the air in the air
chamber, together with the respiration by means of the allantoic vessels. Stimulation
of the respiratory centre by the increasing CO
2
pressure in the blood may be the explan
ation, but there are some facts which are not in accordance with this supposition.
First of all, alteration of the CO
2
pressure in the air chamber does not alter the
frequency of respiration worth mentioning. The following experiment was carried out
with an egg during the parafoetal period.
Egg nr. 5, breeding time 20 days, 5 hours; respiratory movements optically recorded,
freq. 52/min. Air chamber ventilated with room air of 40 C for 5 minutes. After ventilation
the frequency of respiratory movements was 48/min. (no apnoe I), 20 minutes after stopping
the ventilation of the air chamber the CO
2
content of the gas in the air chamber
took 4.10% and the respiratory rythm was 50/min.
The respiratory movements did not alter in rythm, though the CO
2
pressure of the
aspirated air decreased from about 40 mm Hg to about zero and afterwards increased
from zero to the original level. The reverse may be said about the oxygen pressure.
It seems probable that the chemical regulation of respiration in the chicken at this
stage of incubation is only poorly developed. We must however state that the exact
magnitude of the tidal air could not be measured with our technique of registration and
our conclusions bear relation only to the frequency of respiratory movements.
The insensibility of the respiratory centre to chemical stimulation from the aspirated
gases is illustrated by some other experiments.
Egg nr. 6, in the small incubator, incubated 20 days; respiratory movements had a
frequency of 68/min. A mixture of air and 6% CO
2
was driven through the small incubator
(volume = 2 liter), replacing the normal air in it. The respiratory rythm of the animal
did not alter during this experiment and after ventilating the incubator with 8 liters
of the gas mixture (for 20 minutes) the frequency of respiratory movements was only
72/min. [\ow, the gas volume of the incubator was quickly replaced by fresh room air
(in 4 minutes) and the respiratory rythm continuously recorded. No alteration could
put on record, the frequency remained 68/min. and did not change before the animal
perforated the hard egg shell.
Perforation of the shell is another important moment in foetal development, especially
when the small window in the shell is large enough to allow a gas exchange between the
air chamber and the incubator. A sudden fall in CO
2
content and consequent rise in O
2
content of the gas in the air chamber must result and when the chemical regulation of
respiration in the embryo should have its full development an apnoe would occur. To
check this supposition, the small window made by the embryo was enlarged artificially
immediately after perforation, and the respiratory movements of the animal observed
with the eye. In no case apnoe could be stated, the respiratory rythm was about the
28 c. ROMIJN: PARA FOETAL RESPIRATORY MOVEMENTS OF THE CHICKEN
same as before shell perforation, though general movements of the neck and other parts
of the foetus were less frequent during some time.
We put another egg in an atmosphere of 40 C, containing 7% CO
2
, 12% O
2
and
81 % N
2
, a few minutes after perforation of the shell by the embryo. Hatching stated quite
normally, the chicken was completely free from the shell about 3 hours after the first
perforation and the respiratory movements had a frequency of fi8/min. shortly after
birth. Brought under normal conditions the respiratory rythm did not alter and was fixed
at GO/min. for about 15 minutes. When the animal is put back into the first gas mixture
the respiratory frequency does not change, no dyspnoe was observed. The same chicken,
24 hours old, placed in an atmosphere containing 7% CO
2
, 12% O
2
and 81 % N
2
, produces
a distinct dyspnoe and the conclusion seems justified that C O ~ has no effect on respiratory
activity of the chicken during the parafoetal period and during the first hours after birth.
To what extent the low oxygen percentage of the air chamber gas will be of importance
for the development of anoxaemia, consequently for the normal development of the
whole animal in general, is the subject of further research. It will be of importance
in this connection to know the influence of CO
2
on the O
2
combining power of the
chicken's blood.
Moreover it is noteworthy to mention the redundance of the curious gas phase in
the air chamber for normal developm('nt. From three eggs, the air chambers of which
were opened on the 12th day of incubation, two produced a normal chick; removal of the
shell from the air chamber on the 17th breeding day in three other eggs did not prevent
normal hatching. The chickens have grown up in the laboratory to normal 'White Leghorn
hens.
SUMMARY.
The respiratory movements of the chicken during the last day of incubation (parafoetal
period) have been recorded optically. The changes in respiratory activity before and after
the shell perforation have been described and the influence of CO
2
on it discussed.
RESUME.
Les mouvements respiratoires du poussin pendant Ie dernier jour de la couvaison
ont etc ctudics. L'activite de la respiration avant et apres la perforation de la coquille
et l'influence du CO
2
ont ete discutees.
LITERATURE.
ROMIJN, C. & Roos, J., 1938. The air space of the ben's egg and its changes during the period of in
cubation. J. Physiol., 94: 365-379.
WINDLE, W. F. & BARCROFT, J., 1938. Some factors governing the initiation of respiration in the chick.
A mer. J. Physiol., 121: 684-699.