Emphysema Produced in Dogs
by Cigarette Smoking
Oscar Auerbach, MD, E.
David Kirman, and Lawrence
Ten dogs smoked cigarettes daily in two sessions each
day by voluntary inhalation through a tracheostomy tube.
Five dogs died during the course of the experiment, and
the remaining five were killed after more than 420 days
of smoking. Hematocrit values increased markedly during
the first several weeks of smoking and then declined
somewhat but remained higher than presmoking levels.
The heart weight relative to body weight was markedly
higher in the ten smoking dogs than in the ten control
dogs. Pulmonary fibrosis and emphysema, similar to
those conditions in human beings, were found in all five
of the smoking dogs killed. No such lung parenchymal
changes were found in ten control dogs.
Thispreliminary
study
mum number of
initially
was
experiment
cigarettes
undertaken simply as a
to determine the maxi¬
which dogs can smoke
daily without showing evidence of severe acute
effects of nicotine or carbon monoxide poisoning.
We were not interested in dosage levels, relative to
body weight, much beyond that taken by some
persons who smoke very heavily. When it was found
that dogs could tolerate a fairly high exposure, the
experiment was continued for more than a year to
get some idea of the long-term effects. When two
dogs died with such massive lung infarction that
the tissue could not be evaluated for structural
changes, the remaining dogs were killed. Causes of
death, lung parenchymal changes, hematocrit and
hemoglobin changes, and heart weights will be dis¬
cussed here. A microscopic study of the bronchial
tubes of the dogs will be reported later.
Method
Tracheostomy was performed on each dog, and
three weeks were allowed for recovery. The trache-
From the Senior Medical Investigator Laboratory, Veterans Ad-
ministration Hospital, East Orange, NJ (Dr. Auerbach and Mr.
Kirman), and the Statistical Research Section, American Cancer
Society, New York (Dr. Hammond and Mr. Garfinkel).
Read before a joint meeting of the sections on diseases of the
chest, anesthesiology, radiology, and preventive medicine and the
American College of Chest Physicians with the General Scientific
Assembly at the 115th annual convention of the American Medi-
cal Association, Chicago, June 27, 1966.
Reprint requests to VA Hospital, Tremont Ave and Center St,
East Orange, NJ 07019 (Dr. Auerbach).
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Cuyler Hammond, ScD,
Garfinkel, MA
ostoma was kept open with a specially designed
hollow tube made of Teflon. In smoking, the tube
was changed to a Teflon tube equipped with a
socket for coupling to a machine designed to de¬
liver cigarette smoke intermittently. The first one
or two puffs (to light the cigarette) were delivered
under forced draft. Thereafter, the cigarette was
smoked by deep voluntary inhalation of the dog.
In between each five puffs (or more often when
necessary to prevent anoxia) the smoking tube was
clamped off, allowing the dog to breathe fresh air
through its mouth or nostrils. Whenever a dog
showed signs of distress, a longer period of air in¬
spiration was permitted.
Initially, the dogs showed high excitability and
symptoms such as occur in a child attempting to
smoke his first cigarette (slight to profuse saliva¬
tion, coughing, dilatation of pupils, redness and
tearing of the eyes, and sometimes nausea, vomit¬
ing, and dizziness). These symptoms subsided sub¬
stantially in subsequent day but reappeared from
time to time, the dogs being variable in this respect.
The dogs were cooperative and, after a week or two,
some showed evidence of liking cigarette smoking
as indicated by tail wagging and jumping into the
smoking box voluntarily.
The plan was to increase daily cigarette con¬
sumption gradually, cutting back to a lower level
whenever it appeared that we might have exceeded
tolerable limits as judged by the reaction of the
dogs (ie, the appearance of acute symptoms more
severe than those experienced by beginning human
cigarette smokers). One female and nine male
beagles of pedigreed stock were selected as experi¬
mental animals. They were divided into two groups,
A and B, the two lightest dogs being put in group
A and the two heaviest put into group B (Table 1).
On four successive days, each dog was put in the
smoking apparatus in the morning and again in the
afternoon, all procedures being carried out except
that the cigarette was not lighted. On the morning
of the next day, (which we will call day 1) each
animal was given one lighted filter-tipped cigarette
of a brand reported to be low in tar and nicotine
content, and on the afternoon of that day was
 Table 1.—Characteristics of the Ten Smoking Dogs
Weight, kg (lb) Mean Hematocrit (%)
Days From No. of
Dog No. Age at Start to Cigarettes At At Before Days Days Highest Last
and Sex Start, Mo Death" Smoked Start End Smokingt 7-1 It 16-20! 2 Weeks? 2 WeeksS
Group A
29 M 19 24 D 73 10.2 (22.5) 10.2 45.9 50.2 51.8 51.8
30 M 19 229 D 1,378 7.6(16.8) 7.5 (16.5) 47.8 55.7 55.3 65.3 59.4
25 410 D 3,539 11.6(25.5) 10.9 (24) 49.1 60.1 58.1 67.4 53.4
36 M 30 415 D 3,598 10.7 (23.5) 11.3 (24.8) 46.8 57.9 65.3 68.1 48.2
37 M 423 K 3,702 11.1 (24.5) 11.9(26.3) 49 50.5 55.9 59.5 56.6
Group B
15 M 21 422 K 4,114 16.3 (36) 15.1 (33.3) 49.3 57 58.1 66.4 51.4
26 M 423 K 4,084 12.8 (28.3) 14.7 (32.3) 48.9 52.1 55.2 64.4 53.7
31 M 17 421 K 4,104 12.6(27.8) 11.9 48.4 56.9 58.6 69.5 57.7
34 M 18 422 K 4,116 10.2 10.4 (22.8) 47.8 56.1 59 66.5 51.8
35 M 278 D 2,298 11.6 11.4(25) 48.5 52.1 55.2 55.5
*
D indicates died: K indicates killed.
+ Mean of readings on four successive days before smoking.
Î Mean of readings on five successive days (smoking cigarettes).
§ Mean of readings taken during a period of two weeks.

Table 2.—Daily Number of Cigarettes

the daily cigarette smoking of the remaining nine
Smoked by Dogs in Groups A and B dogs for several days. On day 31, cigarette con¬
No. of Cigarettes sumption was increased but was reduced again on
From Start
Day Smoked per Day day 38 due to acute symptoms in several dogs.
of Smoking Group A Group B Later, consumption was increased again without
1-2
3-10
1
2
1
2
appearance of acute symptoms.
11 2 4 It should be noted that Table 2 shows the gen¬
12-15 3 4 eral schedule of dogs in groups A and B. Occasion¬
16-20 4 6
21-23 6 8 ally, owing to symptoms, an individual dog was
24 5 7 given a temporary reduction in smoking. For ex¬
25-29
30
3
4
5
6
ample, on two occasions dog 35 was temporarily
31-37 5 7 taken off smoking and given antibiotic therapy for
38-106 4 6
7
high fever and apparent pulmonary infection. This
107-120
121-148
5
6 8 dog had to be handled carefully, often being given
149-162 7 9 fresh air between puffs of cigarette smoke, and he
163-176
177-190
8
9
10
11
appeared not to inhale the smoke quite as deeply
10 12 as most of the other dogs. On day 24 convulsions
191-204
205 207 11 13 developed in dog 31. Smoking was immediately re¬
208-226 10 12 duced but later resumed at the level of other dogs
227-423 12 12
in group B.
Five of the smoking dogs died during the course
given one unlighted cigarette. This was repeated of the experiment, and the remaining five were
on day 2. On days 3 to 6 one lighted filter-tipped killed on days 421, 422 and 423.
cigarette was given in the morning and one in the Ten additional dogs of the same breed and of
afternoon. On days 7 to 10, two cigarettes of a approximately the same sizes and ages as the ten
popular 70-mm non-filter-tipped brand were given, smoking dogs were kept as controls. Two of them
one in the morning and one in the afternoon. Ciga¬ had tracheostomy openings. They were not exposed
rettes of this brand (which was reported to be to cigarette smoke but were killed at the time of
relatively high in nicotine and tar) were used there¬ death of the last five smoking dogs.
after. Each cigarette was smoked down to a re¬
maining butt length of approximately 5 mm. Results
Daily dosage throughout the remainder of the Food Intake and Body Weight.—Food intake and
experiment is shown in Table 2. There were two body weight were determined daily. The appetites
smoking sessions per day, morning and afternoon, of the smoking dogs remained excellent although
seven days a week. On days when the dogs smoked this varied somewhat from time to time. As indi¬
an odd number of cigarettes, one more cigarette cated in Table 1, none of the dogs showed a marked
was given in the morning than in the afternoon. change in weight up to the time they died or were
The dogs were fed once a day about half an hour killed.
after the afternoon session. As shown in Table 2, Hematocrit.—~Ear blood samples were taken from
up until day 227 dogs in group A were given fewer the smoking dogs on four successive days before
cigarettes per day than dogs in group B. the start of smoking and once a day immediately
Several hours after the morning smoking session following the morning smoking session on days 1
on day 24, an apparently healthy dog which ap¬ through 26. Thereafter, ear blood was drawn every
peared to have good tolerance for cigarette smok¬ Tuesday and Friday immediately after the morning
ing was found dead in its cage. Fearing that we smoking session. Ear blood samples were similarly
might have exceeded tolerable limits, we reduced drawn from five of the nonsmoking control dogs on

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 1. Lung parenchyma of nonsmoking dog 38 (top) and
of smoking dog 26 (bottom). Dog 26 had smoked 4,084
cigarettes in 423 days (x40).
Tuesday and Friday mornings during 27 consecu¬
tive weeks. Of these five nonsmoking dogs, two had
tracheostomas and three did not.
Microhemátocrits were determined by the stan¬
dard capillary tube technique.1
The mean hematocrit readings of the five control
dogs were 47.7%, 47.9%, 45.3%, 50.7%, and
48.7,% respectively. This is in agreement with a
previous report on normal male beagles.2 Although
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2. Lung parenchyma of nonsmoking dog 38 (top) and
of smoking dog 26 (bottom) at higher magnification
(X150).
the hematocrit readings for individual dogs varied
from time to time, there was no appreciable trend
either up or down during the 27 weeks of observa¬
tion. The highest single hematocrit reading for any
of these five dogs was 55.5%. As shown in Table 1,
the mean hematocrit values of the ten smoking
dogs on the four days preceding smoking ranged
from 45.9% to 49.3%, this being in the same range
as that of the control dogs. Of the 40 individual
 3. Lung parenchyma of nonsmoking dog 42 (top) and
of smoking dog 34 (bottom). Dog 34 had smoked 4,116
cigarettes in 422 days (X40).
readings taken during this period (four readings
on each of the ten dogs), the highest was 51%.
The hematocrit values of all ten experimental
dogs began to rise when they started to smoke. The
mean reading for each of the ten dogs on days 7 to
11 and on days 16 to 20 is shown in Table 1. For all
ten dogs, the mean hematocrit value during days 7
to 11 was higher than the before-smoking level and
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4. Lung parenchyma of nonsmoking dog 42 (top) and
ofsmoking dog 34 (bottom) at higher magnification
(X150).
in eight of the ten dogs it was still higher during
days 16 to 20. Following this, the readings fluctu¬
ated considerably but showed a tendency to rise in
all ten dogs. The highest mean of any four readings
taken consecutively during a two-week period
varied from 51.8% (dog 29) to 69.5% (dog 31). In
dog 36 the highest mean level occurred during the
fourth and fifth weeks of smoking, while in most of
 the other dogs it was not reached until the 12th to
14th week of smoking. In later weeks, despite an
increase in daily cigarette consumption, the mean
hematocrit readings tended to decline, the degree
of decline varying in different dogs (last column,
Table 1 ). The highest single hematocrit reading for
each of the ten dogs was 57% for dog 29, 68% for
dog 30, 71% for dog 32, 69.5% for dog 36, 68% for
dog 37, 71% for dog 15, 67% for dog 26, 71% for
dog 31, 72% for dog 34, and 64% for dog 35.
Cause of Death.—Dog 29 died unexpectedly on
day 24. At autopsy, a large recent thrombus was
found in the right auricular appendage. In addi¬
tion, several recent infarcts measuring from 5 mm
to 2 cm were found in both lungs.
Dog 30 died suddenly on day 229 while smoking
the fourth cigarette of the afternoon session. He
had shown signs of ill health during the preceding
several days. Autopsy indicated recent broncho-
pneumonia (with no evidence of abscess formation)
as the cause of death.
Dog 35 died on day 278 while inhaling smoke
from the third cigarette of the afternoon session.
He had appeared to be in fairly good health during
the preceding several weeks. At autopsy, thrombi
were found in both lungs, with areas of infarction.
Dog 32 was found dead early on the morning of
day 410. She had had wheezy, labored respiration
for several weeks, and her movements were slow
with appearance of fatigue. Rectal temperatures
taken frequently during this period were normal.
At autopsy a recent thrombus was found in the
right auricular appendage and emboli with massive
infarction were found in both lungs.
Dog 36 was found dead on the morning of day
415. The animal had been ill for several days with
a high fever and was under treatment with anti¬
biotics and intravenously administered fluids. At
autopsy, a large recent thrombus was found in the
right auricular appendage and there was massive
infarction of both lungs, with secondary pneumonia.
We do not know why more dogs died in group A
than in group B. Perhaps it may be related to the
body weight of the animals at the start of the
study. The dogs in group A were lighter than those
in group B.
Autopsy Findings
Lung Parenchyma.—All of the lungs were filled
with formaldehyde solution instilled into the
trachea soon after removal from the body. The
lungs of beagle dogs have seven lobes, and a speci¬
men of parenchymal tissue was taken from each
lobe for microscopic study.
Grossly, the lungs of all ten nonsmoking, con¬
trol dogs showed a pink surface, as did the cross
sections of the lungs. Microscopically, the lung
parenchyma of these dogs showed a uniform ap¬
pearance strikingly similar to that of a human
being who has never smoked and never been ex¬
posed to air pollutants3 Fig 1 to 4). The sizes of
the alveolar spaces were within normal limits and
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the alveolar septa were thin. In none of the ten
control dogs was there evidence of pulmonary fibro¬
sis or rupturing and tearing of alveolar septa.
As previously noted, five smoking dogs died dur¬
ing the course of the experiment and five were killed
after 421, 422, or 423 days of cigarette smoking.
In dog 29 (death on day 24), pad-like attach¬
ments to alveolar septa were found in all lobes of
the lungs. Dilatation of the alveolar spaces was
present in focal areas within the lungs. These were
located chiefly in the subpleural aspect of the lungs.
After 229 days of smoking (dog 30), the changes
in the lung, although focal, were more advanced.
The changes were present predominantly in the
subpleural regions. Here the alveolar septa showed
a fibrous thickening of the walls with areas of rup¬
ture.
The lung of dog 35 who died after 278 days of
smoking showed focal subpleural pulmonary fibro¬
sis and ruptured alveolar septa but to a lesser de¬
gree than dog 30. This was the dog who had to be
taken off smoking occasionally and who appeared
to inhale the smoke less deeply than other dogs.
Dog 32, who died after 410 days of smoking, and
dog 36, who died after 415 days of smoking, showed
such extensive infarction of the lung parenchyma,
that it was not possible to evaluate the extent of
pulmonary fibrosis and tearing of alveolar septa in
the massive necrotic tissue.
The findings in the lungs of the remaining five
dogs, which were killed after 421, 422, or 423 days
of smoking, showed a fairly constant picture on
gross and microscopic examination.
On gross examination the surface of the lungs
showed white areas which varied in size from 5 mm
to 3 cm. These were present over all the lobes but
were generally largest in the apical lobes. The re¬
mainder of the surface had a pink appearance.
Cross section of the lung parenchyma showed the
presence of dilated air spaces, particularly beneath
the pleural surface. These measured 1 mm to 1.5
mm in size and gave the lung in these areas a
spongy appearance.
Microscopically, the white areas showed irregu¬
larly arranged zones of connective tissue surround¬
ing dilated airsacs of varying size within many of
which remnants of alveolar septa were still present
(Fig 1 to 4). In other regions of the lung paren¬
chyma, fibrous thickening of the alveolar septa and
dilated air sacs, although present, were not of an
advanced degree. There was no thickening of the
walls of small arteries and arterioles within the
lung.
Figures 1 to 4 show photomicrographs of the lung
parenchyma of smoking dogs 34 and 26. These
were selected for presentation as typical of the
changes found in the five smoking dogs killed at the
end of the experiment. Photomicrographs of the
lung parenchyma of two nonsmoking dogs are
shown for comparison. Nonsmoking dog 38 had a
tracheostoma and nonsmoking dog 42 did not.
The lung parenchyma of a number of the smok-
 ing dogs showed granulomata of varying sizes.
Some of these contained fat, others contained
brown pigment, and still others contained both fat
and brown pigment. In one of the smoking dogs
brown pigment was also present in the regional
lymph nodes. No granulomata were found in the
lungs of the nonsmoking control dogs.
Heart Weight.-The mean heart weight expressed
in grams per kilogram of body weight was 7.87 for
the ten control dogs, the individual readings being
6.61, 6.98, 7.09, 7.23, 7.34, 8.08, 8.61, 8.83, 8.96,
and 8.99. This is consistent with typical values for
normal male beagles as reported by Hadidian and
Pawlowski.a The mean for the ten smoking dogs
was 11.94, the individual readings being 13.22,
15.95, 12.66, 11.37, 8.88, 13.62, 12.01, 9.51, 9.47,
and 12.69, respectively, in the order in which the
dogs are listed in Table 1. The difference of 4.07 in
mean values between the ten normal dogs and
the ten smoking dogs is statistically significant
(P< 0.001).
Comment
Hematocrit studies were made on the dogs be¬
cause in a previous experiment on human beings,
cigarette smoking was found to be associated with
a marked increase in hematocrit, and the authors
suggested that this might possibly be related to
"the occurrence of diseases characterized by throm¬
bus formation."4 Hematocrit values of the dogs
increased with smoking (though later they declined
somewhat), and several of the dogs died of throm¬
bosis with pulmonary emboli and infarction.
Whether these two events were causally related is a
matter of conjecture.
Prospective epidemiological studies have indi¬
cated a high degree of association between cigarette
smoking and death rates from pulmonary emphy¬
sema.5,6 In a histologie study of the lung parenchy¬
ma of 1,340 men who died of various causes, we
found a high degree of association between ciga¬
rette smoking and rupturing of alveolar septa to¬
gether with fibrosis.:> Similar findings have been re¬
ported in a study of macrosections of human
lungs.7
References
1. Deubler, J.J., and Cole, E.J.: The Microhematocrit in the
Routine Hematologic Examinations of Dogs, N Amer Vet 38:367\x=req-\
377 (Dec) 1957.
2. Hadidian, Z., and Pawlowski, A.A.: Hematologic, Biochemi-
cal and Organ Weight Characteristics of Beagles, Cancer Che-
mother Rep 38:17-23 (May) 1964.
3. Auerbach, O., et al: Smoking Habits and Age in Relation to
Pulmonary Changes, New Eng J Med 269:1045-1054 (Nov 14)
1963.
4. Eisen, M.E., and Hammond, E.C.: The Effect of Smoking on
Packed Cell Volume, Red Blood Counts, Haemoglobin and Plate-
let Counts, Canad Med Assoc J 75:520-523 (Sept 15) 1956.
5. Hammond, E.C.: Smoking in Relation to the Death Rates of
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Holland et al8 have reported the occurrence of
emphysema in rabbits exposed to cigarette smoke
in an exposure chamber daily for 2 to 5% years.
Hernandez et al9 exposed greyhound dogs to ciga¬
rette smoke in an exposure chamber five times a
week twice daily for periods up to more than one
year and reported marked changes in lung paren¬
chyma related to the duration of exposure.
Instead of using an exposure chamber, we tried
to simulate human smoking by having the dogs
draw smoke directly from cigarettes into their
lungs during two sessions each day, seven days per
week. Daily consumption of cigarettes was grad¬
ually increased to the number smoked per day by
some persons who are extremely heavy smokers,
body weight being taken into consideration. Ad¬
vanced changes were found in the lung parenchyma
of every one of the five dogs who survived exposure
for more than 420 days and were then killed. No
such changes were found in the lung parenchyma
of ten unexposed control dogs.
The lung parenchymal changes in the smoking
dogs showed the characteristics of emphysema
found in persons who smoke heavily, ie, rupturing
of alveolar septa, fibrous thickening of alveolar
septa, and pad-like attachments to alveolar septa.
In human beings who died at an advanced age
after having been heavy cigarette smokers for many
years, we found marked thickening of the walls of
arterioles and small arteries in the lung parenchy¬
ma.4 This particular type of change was not found
in the lung parenchyma of the smoking dogs. This
may have been due to the fact that the dogs were
younger at death (relative to the normal life ex¬
pectancy of the species) than were the human cig¬
arette smokers in whom these changes were ob¬
served.
Heart weight per kilogram of body weight was
found to be greater in the smoking dogs than in the
control dogs. Perhaps this was secondary to lung
parenchymal changes, as in cor pulmonale, but this
is not necessarily the correct interpretation.
This investigation was supported in part by grant P-315 from
the American Cancer Society.
Compilation and statistical analysis of data were done by the
Statistical Research Section of the American Cancer Society.
1 Million Men and Women, Nat Cancer Inst Monogr 19:127-204
(Jan) 1966.
6. Kahn, H.A.: The Dorn Study of Smoking and Mortality
Among U.S. Veterans: Report on 8 1/2 Years of Observations, Nat
Cancer Inst Monogr 19:1-125 (Jan) 1966.
7. Anderson, A.E., Jr., et al: Emphysema in Lung Macro-
sections Correlated With Smoking Habits, Science 144:1025-1026
(May 22) 1966.
8. Holland, R.N.; Kozlowski, E.J.; and Booker, L.: The Effect
of Cigarette Smoke on the Respiratory System of the Rabbit: A
Final Report, Cancer 16:612-615 (May) 1963.
9. Hernandez, J.A., et al: Pulmonary Parenchymal Defects in
Dogs Following Prolonged Cigarette Smoke Exposure, Amer Rev
Resp Dis 93:78-83 (Jan) 1966.