ANAB&OBIC ORGANISMS IN ACUTE AID CHRONIC

P0LMOHART DISEASES

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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

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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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13

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