anab&obic organisms in acute aid chronic p0lmohart diseases/67531/metadc... · anab&obic...
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ANAB&OBIC ORGANISMS IN ACUTE AID CHRONIC
P0LMOHART DISEASES
APPRO?® 5
I Jor professor
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Director of the Departmentof Biology
Dean of'tbi ''cStra&al® lefcool1
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H S S X B
P r e s e n t e d t o t h e G r a d u a t e C o u n c i l o f t f e *
n o r t h T t t u B t * t * C « l l « g « l a P a r t i a l
F m l £ i l l u # » t o f t h e R « q u i r « * M i t »
F o r t l i # B « g r « « o f
M A S S ! Of i m i
Br
O « o r g « Btiffr *1<M»1 # B* S «
D « a t * a # T e x a s
A u g u s t , I f
TABU OF CONTENTS
Page
LIST OF TABUS iv
Chapter
I. INTRODUCTION 1
Review of Literature Th© Problem
II. MATERIALS AND LABORATORY WORK 4
Materials Laboratory Work
III. EXPERIMENTAL DATA AND DISCUSSION ?
Experimental Data
If. MTI®iA!! AND CONCLUSIONS 32
Stissiary Soael«»i@s8
BIBLIOORAPII 34
lit
LIST OF TABLES
Table Pag®
I. Patients Diagnosed as Having a Bulraonary Infection and Admitted to the Hospital • • • 8
XI* Patients Admitted to the Hospital With Some Condition Other Than Pulmonary# However, at the Time of Admittance or During the Convalescent Period Contracted a Pulmonary Ailment of a Chronic Type » • • • • • • • • Id
III* Patients Seen in an Out-Patient Clinic Stff£eri'ng With a Chronic Type' ©f Pulmonary ' Infection 25
iv
CHIPfH I
INTRODUCTION
Sinee a great deal of time 1® involved in isolating
strict anaerobic bacterial organisms, these, with reference
to their clinieal importance In diseases of the respiratory
tract, have been ignored generally. Most of the anaerobic
organisms which have been studied clinically have been isolated
from gangrenous material, abscesses, septicemias, and cases
of puerperal sepsis., this study has been devoted to a deters
mination as to whether anaerobic organisms are involved in
pulmonary diseases, particularly those of the chronic type*
Review of Literature
An interest in anaerobic organisms of the upper respir-
atory tract began about 1927 when Branham (1} isolated seven-
teen strains fro® nasopharyngeal washings of twenty-six
persons#
Long and Muellerschoen (4) in 1933 substantiated a pre-
vious work which showed that gram-negative anaerobic bacterial
organisms constitute a part of the noma! flora of the naso-
pharynx. However, they found that these organisms are more
often characteristic ©f those patients who suffer more fre-
quently from upper respiratory tract infections, fhey believed
that under proper conditions these anaerobic organisms became
opportunists and assumed a pathogenic condition*
Cohen (2I la 1933 isolated a number of anaerobic bac-
terial organisms from patients suffering from tog abscesses.
During the same year Cohan (3) reported that 80 per cent of
the gram-negative rod-shaped organisms found in lung ab-
scesses gave a positive Shwartzman Phenomenon reaction em-
ployed for the presence of toxic factors#
The Problem
The purpose of this investigation was to determine
whether anaerobic bacterial organisms could be fottod in
chronic pulmonary diseases as well as in acute respiratory
infections. The problem has consisted of, first, the collec-
tion of sputum samples from chronic pulmonary cases; second,
the inoculation of these samples into both aerobic and
anaerobic culture media; third, the determination of the
presence or absence and number, if present, of anaerobic
organisms; and finally, an evaluation of the results obtained
with respect to the problem involved*
BIBLIOQRAPHX - CHAPTER I
1. Braaham, S. H,, "Anaerobic Microorganisms in Naso-pharynx Washings," Journal of Infectious W H I (1927), 203-207, ' • "
2# Cohen, «J#, "Anaerobic Gram-Negative Bacilli Isolated rroia Abscess of the Lung.11 Journal of Infectious Mseaaea. LII _ (1933), 1*5-91* "
3. Cohen, J., "The Bacteriology of Abscess of the Lung and Methods for Its Study"1* Archives of Sura try. XXI? (1932), m-as. 1 ; —
4. Long, P. H. and Kuellerschoen, B.# "Incidence of Anaerobic, FiIter-Passing Organisms in Normal and Abnormal Respiratory Tract Conditions." Journal Diseases. LII (1933), 121-25*
CHAPTER II
MATERIALS AMD LABORATORY WORK
Materials
Materials used in pursuing this problem were cardboard
sputum containers, trypsin, tryptose broth, thioglycollate
broth, rabbit*s blood agar plates, and a Brewer Anaerobic
jar. These were obtained from the Bacteriology Laboratory and
Stock loom of Parkland Meoorial Hospital, Dallas, Texas*
Laboratory Work
Sterile cardboard sputum containers used in the collection
of sputtm. specimens were distributed t© the ysrioui nurses*
stations in the hospital• Upon receipt of the collected
specimens in the hospital bacteriology laboratory, trypsin
was added to each immediately in order to digest the protein*
Elimination of such extraneous material facilitated the pro-
curement of a more satisfactory and representative sample for
the later inoculations. Each sputum specimen was inoculated
into both tryptose and thioglycollate broth for the routine
culture study* In addition one rabbit*® blood agar plate
was inoculated for the culturing of anaerobic organisms.
The materials set up for aerobic cultures were incubated
at 37° Cm for twenty-four hours and then examined for growth#
Tentative identifications of the aerobic organisms were made
4
and the quantity of growth was recorded as slight, moderate,
or heavy# This information was used further la a comparison
®f the growth her© with that ©a the corresponding anaerobic
Mood agar plate# Eaeh different type colony which was sus-
pected of being pathogenic was picked and streaked to another
blood agar plate for aerobic culture. This was done in order
to obtain a pure culture for identification and antibiotic
sensitivity study.
The properly inoculated anaerobic blood agar plates were
placed is a Brewer Anaerobic Jar* The jar was evacuated of
air by a vacuum pump and refilled with methane gas. This
operation was repeated three consecutive times or until the
blood plates assumed a cyanotic condition# The container of
anaerobic plates was placed in an incubator at 37° G» for a
minimum of 3even and a maximum of ten days. Twice each day,
once in" the morning and once in the evening, the blood agar
plates were examined through the Brewer Jar in order to see
if they had retained their cyanotic condition*
At the and of the incubation period the anaerobic plates
were examined and compared with the corresponding aerobic
plate® in order to see if anaerobic organisms were actually
present# The colonies which were suspected of being anaerobic
were picked and sub-cultured on each an aerobic and another
anaerobic blood plate# If the organisms grew on the aerobic
sub-cultured plate this eliminated any possibilities for
their being strict anaerobes. However, if growth occurred
only on the anaerobic subculture this would indicate the
presence of a strict anaerobic organism* To eliminate any
doubt whatsoever the presumptive anaerobic organis® was
again swbettltmred to another aerobic blood agar plate* if
a© growth occurred this time the organism was pronounced
a strict anaerobe#
During the course of this investigation, in west case®
at least, if anaerobic organisms were present a very character-
istic putrid ©dor was noted* Furthermore, most of the
anaerobic organisms encountered developed a coal black-
colony# this facilitated markedly the task of each identi-
fication.
/
CHAPTER III
EXPERIMENTAL DATA AID DISCUSSION
Experimental Data
Sputum specimens were collected fro® 120 patients in
Parkland Memorial Hospital,' Dallas, Texas. These patients '
who were suffering from some pulmonary disease, for'labora-
tory diagnostic convenience, were placed in three distinct
groups*, to® first group, which consisted of fifty-two
patients, were admitted to the hospital with a diagnosed
pulmonary infection, la the second group there were thirty
patients who were admitted to the hospital because of some
conditieto other than pulmonary. However, either at the time
of admittance or during the convalescent period these patients
contracted a pulmonary ailment of a chronic type* The third
group consisted of thirty-eight pulmonary cases which were
seen in am out-patient clinic• Members of this group were
suffering from a chronic type of pulmonary infection and had
been diagnosed as having a *pulmonary disease of undetermined
origin."'
An observation of Table I shows that thirty-one of the
fifty-two patients in Group I were diagnosed as having pneu-
monia* Laboratory procedures revealed that out of these
thirty-one pneumonia patients the sputum of seventeen grew
anaerobic bacterial organisms* Among these, there were
7
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thirteen responsible for anaerobic streptococci producing
black colonies, and two anaerobic streptococci producing
gray colonies. A species of bactericides was isolated from
each ©f two patients# Bacillus ramosus wag isolated from
on® patient. Oat species of an anaerobic cocco-bacillus
was isolated from another patient. Twelve of the thirty-one
Group 1 patients were found to possess pathogenie aerobes#
Among this number there were eight showing the presence of
Siplicoccus pneumoniae, one with a coagulase positive
Staphylococcus aureus> one with a Pseudomonas aeruginosa, one
with a PseudoiBonas aeruginosa and a Staphylococcus aureus
combination, and on® with a Streptococcus pyogenes. However,
eight of the twelve patients showed a combination of anaerobic
bacteria along with the aerobes« Anaerobic bacteria were
Isolated from nine of the thirty-one Group 1 patients with-
out finding any trace of pathogenic aerobes# Neither pathogenic
aerobic nor anaerobic bacterial organisms were isolated from
the remaining eleven of the thirty-one patients in Group Z«
Normal flora only was encountered.
Reference to Table I shows that there were four diagnosed
oases of tuberculosis among the patients of Group X. From
one of these a black colony anaerobic streptococcus was i
Isolated, and from one other there was one pathogenic aerobe*
The remainder revealed nothing other than normal flora
Isolates*
17
Still further reference to Table I reveals that there
were four diagnosed case® of chronic pulmonary disease* in
Group I# Laboratory diagnoses showed that two of these pos-
sessed anaerobic streptococci, ©a® of which produced a black
colony while the other produced a gray colony* Each of these
cases showed the presence of pathogenic aerobes combined with
anaerobes* One case possessed an aerobic pathogen while the
fourth showed only normal pulmonary flora*
A final observation of Table I shows that among the
patients of Group I there were three diagnosed cases of lung
abscess, three of bronchiectasis, one of lung congestion,
one of pharyngitis and asthma, and one of a non-specific
cough* Two of the cases of lung abscess were diagnosed as
hating' anaerobic streptococci, both of which produced black
colonies* Both of these showed additional pathogenic aerobes*
The third case showed only normal flora. Each one of the
three case# of bronchiectasis showed the presence of anaerobic
streptococci, all of which produced gray colonies* Two of
these cases showed a combination of aerobic pathogens with
the anaerobes# The one case of lung congestion grew a gray
colony producing anaerobic streptococcus with combined nor-
mal flora*
An observation of Table II shows that the patients in
Group II had been diagnosed as four hypertensive cardio-
vascular disease, four deferred heart failure, two congestive
heart failure, three cirrhosis of the liver, two
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which a combinatl©a of anaerobic and aerobic organisms were
isolated.
An inspection of Table III reveals that the thirty-
eight patients in Group III had been given a diagnosis of
undetermined origin. However, all appeared to have some
respiratory infection at the time of the visit to the out-
patient clinic. Laboratory diagnoses of the sputum of
twenty-one of the Group III patients showed the presence of
anaerobic streptococci, nineteen of the black and two of the
gray colony type# Further studies showed that sputum from
seven of the group cultured aerobic pathogenic organisms.
Aaong this number were four with Dlplococcus pneumoniae,
one with Streptococcus pyogenes, one with a coagulase
positive Staphylococcus aureus. and one with ©
. two of the aerobic pathogenic organisms were
isolated in combination with anaerobic streptococci* Sputum
of the remaining twelve cases in this group showed nothing
but normal flora#
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CHAPTER If
SUMMARY M B CONCISIONS
Summary
X# Sputum specimens were collected from 120 patients
in Parkland Memorial Hospital, Dallas, Texas# Each of these
patients had been diagnosed as having some type of pulmonary
infection# Laboratory diagnostic testa were made on each
sputum specimentin order to determine whether anaerobic
bacterial organisms were present#
2. For convenience in making this study the patients
were listed in three groups# •The grouping was determined
according to the diagnoses upon admittance to the' hospital
or out-patient clinic#
3# Studies of Group I, which consisted of fifty-two
patient®, revealed the presence of twenty-six individual®
who possessed anaerobic bacteria# la addition twenty-one
of the group possessed pathogenic aerobes# Fifteen of the
twenty-one patients showed a combination of aerobic and
anaerobic bacteria#
4# Studies of Group II, which consisted of thirty
patient®, showed that the sputum of thirteen of these grew
anaerobic streptococci and' that ten of theae grew aerobic
pathogenic organisms* Among the latter group there were two
31
33
patient* who possessed a combination of aerobic and anaerobic
pathogens*
5. An analysis of sputum specimen® from Group III, which
consisted of thirty-eight patients, showed that twenty-one
of these grew anaerobic and seven grew aerobic pathogenic
organisms. Both aerobic and anaerobic pathogens were
isolated in combination fro® two of these patients#
Conclusion*
from the tabulated results ©f bacteriological diagnoses
©f sputum specimens from 120 patients in Parkland Memorial
Hospital, Dallas, Texas, it can be concluded that anaerobic
bacterial organisms are related to and are possibly re-
sponsible for certain chronic types of pulmonary disease#
Probably these are associated with the fulminating type of
respiratory infection as secondary invaders^
BIBLIOGRAPHY
Iranhaa, S, H,. *Anaerobic mcroorganisms In Naso-Pharynx Washings,* Journal of Infectious XL1 (1927), 203-207.
Cohen, J., "Anaerobic Gram-Negative Bacilli Isolated from ' Absc«»a of tha Lung,» Journal of Infaefeious maaaata.
LII (1933), 1SS-91*
Cohen, J», nTh© Bacteriology of Abscess of tha Lung and Mathods for Ita Study," ArcfaHras of Sur&ery. XXIV il932)» 171-**.
Long, P. H. ,aad Muallaracboan, 8*, «Incid@ne« of Anaerobic, Filter-Passing Organisms in Normal and Abnormal Respiratory Tract Conditions," Journal of |nf^otiou> Diseases. Ill (1933), 121-25*
34