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—_— THE PHILIPPINE 


2 JOURNAL OF SCIENCE 


SECTION B 
TROPICAL MEDICINE 


EDITED WITH THE COOPERATION OF 


E. L. WALKER, S. D.; M. A. BARBER, Pu. D.; A. R. WARD, D. V+ M. 
Committee on Experimental Medicine 


V. G. HEISER, M. D.; W. E. MUSGRAVE, M. D.; B. C. CROWELL, M. D. 
Committee on Clinical Medicine 


R. C. McGREGOR, A. B. 


VOLUME IX 
1914 


WITH 15 PLATES, 24 TExT FIGURES, 1 MAP, AND 7 CHARTS 


MANILA 
BUREAU OF PRINTING 


1914 ASSL SEX 


129562 


eh Me rp ae: . 


ay hee ADA 


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RAR A ACER 

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RIGS 


eaten) T Ce 6 als 1 Ra ee 


CONTENTS 


No. 1, February, 1914 


BARBER, M. A. Cockroaches and ants as carriers of the vibrios of 
LaNSREW IG CLG) [TRE pees a tee NS ee a AI de Pree nie eae Rae 


HEISER, VICTOR G. Reappearance of plague in the Philippines after 
an absence of six years. Brief description of the outbreak, the 
methods used to combat it, and the probable factors in its 


BITCLOCUCUIOMM eter nrc Cee ree een On SRW ey 5 CU eee SS 
One map. 


BOYNTON, WILLIAM HUTCHINS. A preliminary report of experiments 
on the cultivation of the virus of rinderpest in vitro.................... 


BOYNTON, WILLIAM HUTCHINS. An atypical case of rinderpest in a 


(QETRE|T EO) Bike Mie oe PS Nera oa Fae acre eM ia |e Ae A EB 
One chart. 


WARD, ARCHIBALD R., WooD, FREDERICK WILLAN, and BOYNTON, 
WILLIAM HUTCHINS. Experiments upon the transmission of 


PEETICO TS OS filo ee ne ieee tae a ee NNO eee ae ee ON 8 
Two plates and 6 charts. 


WILLETS, Davin G. Intestinal parasitism, particularly entameebiasis, 
in patients of the Philippine General Hospital, Manila, P. I........ 


WILLETS, DAvip G., with the codperation of STANLEY, CourT R., and 
GUTIERREZ, PERPETUO. Preliminary report on the treatment of 
entamcebiasis with ipecac, emetine, and neosalvarsan at the Phil- 
mppine: General Hospitals Manila, Pr Wonca. .-cncscsennmsauneccconncecseoee 


GiBson, R. B., and CoNcEPcION, ISABELO. Nerve degeneration in 


Towlsited) on) unhusked rice .(palay))\-2 2212. 2-ctssenactencecceelSonscodek otenfeecses 
One plate. 


WARD, ARCHIBALD R., and Woop, FREDERICK WILLAN. Simultaneous 
method of inoculating cattle and carabaos with serum from ani- 
mals that have been recently immunized .................22........0.2002000200---+ 


Three plates. 
No. 2, April, 1914 


A JOINT COMMISSION OF REPRESENTATIVES FROM THE COLLEGE OF 
MEDICINE AND SURGERY, UNIVERSITY OF THE PHILIPPINES; BUREAU 
OF SCIENCE; AND BUREAU OF HEALTH. Sanitary survey of the 
San Jose Estate and adjacent properties on Mindoro Island, 
Philippine Islands, with special reference to the epidemiology of 
PRACT, Ey ee a ee es A ee 

One plate and 2 text figures. 


No. 3, June, 1914 


CALDERON, FERNANDO. Some data concerning the medical geography 
Olu theme bsnitrr pitches Ve ee ee ed a ate ee eta 2 


Page. 


39 


45 


49 


81 


93 


119 


125 


137 


Iv Contents 


ScHOBL, OTTO. The etiology of trichomycosis palmellina in the Phil- 
AP PINE TS LATS Wee ea ae eae ea Ee NA ee nein rs we ete 
One plate. 
JOHNSTON, JOHN A. A contribution to the bacteriology of leprosy. 
Preliminany}mobe: 22..3 See eae ate OLS Oe Be ee 
One plate. 
WILLETS, DAvID G. Intestinal helminthiasis in the Philippine Islands 
as indicated by examinations of prisoners upon admission to Bili- 
bids Prison. Mantle, Pil... a eel Ae oe ee SN eee 


MUSGRAVE, W. E., and Sison, A. G. Bacillary dysentery: The most 
prevalent form in Manila and its treatment..................-........---.-- 


WILLETS, DAvip G. Widal reactions among healthy adult Filipinos... 


BOYNTON, WILLIAM HUTCHINS. Experiments on the cultivation of 
rinderpest virus as described by Baldrey..............----2-..---sc..0--000--0---- 


BOYNTON, WILLIAM HUTCHINS. Kidney-worm infestation of swine 
in the Philippine Islands with special reference to the patho- 
logical \changes! 2:5. ..s.1..-2c.c access geen Decseen od son oe ott eae sce oe 

Three plates. 
LicHT, S. F. Another dangerous jellyfish in Philippine waters....... 


GIBSON, R. B. Some simple laboratory apparatus............-----...-.-..---- 
Three text figures. : 


FRE VIC WS erecta i ae ee eee nee ee ee 
No. 4, August, 1914 


BARBER, MARSHALL A. The pipette method in the isolation of single 
microérganisms and in the inoculation of substances into living 
cells. With a technique for dissection, staining, and other pro- 


cesses carried out under the higher powers of the microscope... 
Two plates and 19 text figures. 


CROWELL, B. C. Notes on the diagnosis of Asiatic cholera at autopsy. 

GUTIERREZ, PERPETUO. Typhoid fever in the Philippines.................... 

FREVIGWS isc neh ch Bek Se a a ea Re cE 
No. 5, September, 1914 


WALKER, ERNEST LINWOOD and BARBER, MARSHALL A. Malaria in 
the Philippine Islands. I. Experiments on the transmission of 
malaria with Anopheles (Myzomyia) febrifer sp. nov., Anopheles 
(Pseudomyzomyia) rossii, Anopheles (Myzorhynchus) barbiros- 
tris, Anopheles (Myzorhynchus) sinensis, and Anopheles (Nys- 
sonaynehns) wimaculatnstet.2656 non acnad ete ee ee 


WILLETS, DAvip G. Malaria in the Philippine General Hospital, 
Manila; PP). during the fiscal jyear 19130 2 eee 


CROWELL, B. C. The chief intestinal lesions encountered in one thou- 
sand consecutive autopsies in Manila.........2.222200020.. eee 


FREVIC WH sorter eee rete ieee eee eS Tb tee SS 


Page. 


219 


227 


233 


241 
253 


259 


269 


291 
297 


303 


307 


361 
367 
379 


381 


441 


Contents 
No. 6, November, 1914 


Ruepicer, E. H. The germicidal power of glycerin on various micro- 
organisms under various conditions.........................-..-- x AO Ree he 
ScHOBL, OTTo. The vitality of the cholera vibrio in Manila waters.... 


“WALKER, ERNEST LINWooD. The morphology of the adults of the 
filaria, found in the Philippe Uslands.2.).2......2..442. cae 


One plate. 


ACOSTA-SISON, HONoRIA. Pelvimetry and cephalometry among Fili- 
FETE SES ae SO sare oS eel ee es oe eso RENT a J OO Re a eee 


GisBson, R. B. A note on the physiological action of the proteoses.... 


GIBSON, R. B., and CoNcEPCION, ISABELO. The lymphagogic action of 
the Philippine mango, Mangifera indica Linnezus........................ 


CONCEPCION, ISABELO. Observations on mango rash................0.--.----.--- ; 


BarBer, M. A. Milk poisoning due to a type of Staphylococcus albus 
occurring in the udder of a healthy cow...............2..-02...2------------ 


LOT OST il paice iat DUA edi Ae Nt EE eH neh ct CN a 


sgl ary: 
aka 
tH iy Lh hat 
‘ is va 


pet 
heat oh wei Ava 
is so oe a 
ee if Ath) Aaies i a re 
ia bhi ver 
Bs v 5 | 


PUBLICATIONS FOR SALE BY THE BUREAU OF SCIENCE, 
ange PHILIPPINE ISLANDS | 


ETHNOLOGY 


A VOCABULARY OF THE IGOROT LAN- 
GUAGE AS SPOKEN BY THE 
BONTOO IGOROTS 


By WaAtrer CLAyron CLAPP 


Order No. 408. Paper, &9 pages, $0.75, 
postpaid, 
The vocabulary is given in igorot-English 
and English+Igorot, 


THE NABALOI DIALECT 


By Oro ScHEERER 
and 
THE BATAES OF PALAWAN 


By Epwarp Y. MILLE 


Order No. 403. Paper, $0.25; half mo- 
rocco, $0.75; postpaid. 

The “Nabalol Dialect (65 pages, 29 

plates) and the Bataks of Palawan (7 

pages, 6 plates) are bound under one cover. 


THE BATAN wet tie AS A Sn 
OF THE GROUP 
) OF PINGUAGES 
By Otro ScHEERER 
and 
“Fy” AND “Vv” IN PHILIPPINE 
LANGUAGES : 
By Caruos Evegerr Conant 
Order No. 407. 


These two papers are issued ‘under one 
cover, 141 pages, paper, $0.80, postpaid. 


THE SUBANUNS OF SINDANGAN BAY 
By Emerson B. CHristie 


Order No. 410. Paper, 121 pages, 1 
map, 29 plates, $1.25, postpaid. 
Sindangan Bay is situated on the north- 
ern coast of Zamboanga Peninsula. The Su- 
banuns of this region were yh by Mr. 
Christie, during two periods of five and six 
weeks, respectively. 


The 29 plates illustrate the Subanuns at ~ 


work and’ at play; their» industries, houses, 
altars, and implements; and ‘the people 
themselves, 


THE HISTORY OF SULU 
By NAJEEB M. 'SALEEBY 


Order No. 406. Paper, 275 pag 4 
maps, 2 diagrams, $0.75, postpaid, 
In. the preparation tel, ls ere for 
The History of tS 


much. time and in gs hs access 


to deouments in the possession of the Sultan 
is a history of the 


of Sulu. . This 
Mores in the Philippines from the earliest 
times to the dd actaae occupation, , 


pe Saleeby spent 


ETHNOLOGY—Continued | 
conlahan 5 Ae St aegis 
By Najnss M. SaLexsy 
Order No. 405. Pabar, 107 pages, 15 


plates, 5 diagrams, $0.25;. half mo- 
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This ‘volume. deals. with ‘the _ earliest. 


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The names of the rulers. of ‘Magindanao are 
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NEGRITOS OF ZAMBALES 
By Wuit1am Avan Resp 


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Plates from photographs, many. of which 
were taken for this publication, show orna- 
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INDUSTRIES 
PHILIPPINE HATS 
By C. B. Roginson 


Order No. 415. Paber, 66 pages, 8 
plates, $0.50 postpaid. 


This paper fs a concise record of the 
history and present condition of hat making 
in the. Philippine Islands. 


THE ya INDUSTRY IN THE 
LAND OF NEGROS” 


By Herperr 8; WALKER 


Order No. 44 Paper, 145 pages, 10 
plates, 1 map, $1.25, postpaid. | 


Considered from the viewpoint of prac- 
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in the Island of Negros is one of the mos 
important papérs published -by the Bureau 
of Sclence.. This volume is.a‘real contribu. 
tion. to the’ subject; it is nota mere com- 
pilation, for the author was in the field and 
be the . conditions» of which he 
writes. : 


A MANUAL OF PHILIPPINE SILK 


By CHARLES §. BANKs 


Vorier ‘No. 413. . Paper, 53° pages, 20 
plates, $0.75, postpaid. 

In A Manual of Philippine Silk Culture » 
are presented the results of several years’ 
actual work’ with at paednar ge larva to 
new cone. 


gether witha desoription Sel 


, bine race. 


THE PHILIPPINE ~S&tiens! sis: 


Too RNAL OF SCIENCE: -.. 


B. TROPICAL MEDICINE 
Vou. IX FEBRUARY, 1914 No. 1 


COCKROACHES AND ANTS AS CARRIERS OF THE VIBRIOS 
OF ASIATIC CHOLERA * 


By M. A. BARBER 


(From the Biological Laboratory, Bureau of Science, Manila, P. I.) 


Cockroaches, especially of the species Periplaneta americana 
Linn., are very common in dwelling houses in Manila at all 
seasons of the year. They are voracious feeders on all kinds 
of organic matter, and at night time, especially, creep over any 
unprotected human food and discharge their feces there. Since 
they have ready access to human feces in closets not provided 
with sanitary plumbing, they may become the carriers of in- 
fective material from the closets to human food. This is the 
more probable, since these insects not only creep rapidly, but also 


fly from place to place. In the following experiments, it was 


purposed to test the feeces of cockroaches which had previously 
ingested cholera cultures and human feces containing cholera 
vibrios, with the purpose of determining if these insects may 
not become carriers of Asiatic cholera. 

The cockroaches used in these experiments were all Peripla- 
neta americana and all winged adults. The method of experi- 
mentation was simple. The insects were caught and kept in a 
jar for a day or more until they had become hungry. They were 
then distributed in wide-mouthed bottles, one insect to a bottle. 
Cultures of cholera or liquid human feces were introduced into 
the bottles by means of a pipette. Some powdered carmine was 
added to the feces or cultures so as to make it possible to identify 
a feces sample with a given feeding. Liquid feces obtained 


* Received for publication December 8, 1913. 
124298 


2 The Philippine Journal of Science 1914 


from cholera patients were used with no addition to make them 
more palatable to the insects. 

The cockroaches devoured these feeces greedily; sometimes, 
a single insect ingested 0.2 cubic centimeter. The insects were 
transferred to new dry bottles after feeding and kept under 
close observation in order to obtain feeces fresh for testing. 
About six hours after their meal, the insects usually began dis- 
charging liquid or semiliquid feces, deeply tinted with carmine. 
On the day following the feeding and on subsequent days, the 
insects were given beef broth containing maltose, but no more 
cholera vibrios or carmine. The maltose broth was fed to them 
in order to obtain fresh feces for testing. A discharge of feeces 
usually followed this meal—often immediately. Carmine some- 
times persisted in the feces for six days after feeding. 

Tests were made by immediate microscopical observation, by 
direct transfer to Dieudonné plates, and by transfer to peptone 
for subsequent test on Dieudonné plates. All vibrios were tested 
by a specific agglutinating serum. 

In 8 cockroaches fed with human feces, cholera vibrios were 
recovered from the insects’ feeces six hours or more after feeding. 
In one case they were found in comparatively small numbers 
seventy-nine hours after feeding, and in several other cases they 
occurred in greater or less numbers from twenty-four to forty- 
eight hours after the ingestion of feeces. In one case they were 
obtained twenty-four hours after feeding with a cholera culture. 

Actively motile cholera vibrios often appeared in enormous 
numbers in the insects’ feces. In two cases, thirty-two hours 
after feeding, they were so plentiful that the material was im- 
mediately suitable for an agglutination test in hanging drop. 
They were apparently as numerous as in the original human 
feces. In another insect, Dieudonné plates made directly from 
feces discharged two days after feeding showed thousands of 
colonies. They were also seen in great numbers in feces at less 
intervals after the feeding. Generally, they seemed to diminish 
in number after twenty-four hours. In two cases, feces ob- 
tained twenty-nine and one-half and thirty and three-fourth 
hours, respectively, after feeding gave negative results, although 
carmine still persisted in the feces and these same insects had 
passed feces containing cholera vibrios five hours previously. 
On the following day, the feces of both these insects were free 
from carmine and gave negative cholera tests. The cockroaches 
were kept at a room temperature of from 29° to 31° C. 

The longevity of cholera vibrios in cockroach feces after dis- 
charge from the insect is probably short when the feces are 


ES a 


3 
q 
‘ 
. 


IX, B,1 Barber: Carriers of Asiatic Cholera B 


deposited in places where they can readily dry, but when dis- 
charged on moist food vibrios may remain for some time. To 
test this matter, some fresh cockroach feces were placed on four 
different kinds of food: fresh beef, lettuce, fish, and clams. In 
every case, the cholera vibrios remained viable for at least sixteen 
hours at room temperature. 

In this connection it may be worth while to mention the results 
obtained by me in some recent experiments on the longevity of 
cholera vibrios in human feces placed on various foods in use 
in this locality. Here, as in the case of the experiments with 
cockroach feces, the cholera vibrios must compete with the other 
bacteria present in the feces and with the microdrganisms 
already present in the food. On the cut surfaces of a cucumber, 
a chico, and a papaya fruit—all distinctly acid to litmus—and 
on the leaves of lettuce, the cholera vibrios survived overnight 
in one experiment. In another test, cholera vibrios in human 
feeces survived twenty hours on the cut surfaces of cucumber 
and papaya fruits, but failed to live forty-four hours. On° 
two varieties of shrimp, cholera vibrios in human feces re- 
mained viable twenty-two hours; on oysters, forty-six hours; 
and on the inside of an opened clam, four days. In all of these 
four foods, other bacteria were very numerous; in the oysters, 
especially, they were so plentiful that the food was acid in reac- 
tion when the cholera vibrios were placed on it. It is probably 
fair to assume that cholera vibrios, when abundant in cockroach 
fzeces, would survive as long on foods as when in human feces. 

In addition to their feeces, the vomit of these insects may also 
be a source of danger. A fzces-fed cockroach was observed to 
disgorge portions of its meal at intervals of ten minutes, twenty 
minutes, and sixty minutes after feeding—sufficient time to allow 
the insects to travel from a closet to human food. The sixty- 
minute sample contained many cholera vibrios. These insects 
often discharge a thin saliva from the mouth. This fluid seems 
to be nearly, if not quite, bacteria-free, and tests failed to show 
any cholera vibrios in it. The vomit mentioned above is of a 
different nature, as shown by the carmine tint and by bacteria 
and other substances seen in microscopical examination. 

As might be expected, cockroaches show no evidence of infec- 
tion by cholera. The vibrios are simply held in the intestine, 
and apparently become no more numerous than in the ingested 
human feces. In order to determine if there is any loss of viru- 
lence in cultures of cholera vibrios isolated from cockroach 
feces, a comparative test was made by guinea-pig inoculations. 


4 The Philippine Journal of Science 1914 


A culture was isolated from a sample of human feces, and this 
sample was then fed to a cockroach. Twenty-nine hours after- 
ward, a second culture was isolated from the fzces of the cock- 
roach. Young agar cultures of the two cultures were tested by 
intraperitoneal inoculation into 200-gram guinea pigs. In the 
culture isolated from the cockroach, one-eighth of a loop was fatal 
after about twenty-seven hours and one-sixteenth of a loop failed 
to infect. In the culture isolated directly from the human feces, 
one-fourth of a loop was fatal in less than twenty-four hours 
and one-eighth of a loop nonfatal. So there is no evidence of 
Joss of virulence in cholera vibrios after twenty-nine hours in the 
antestine of the cocns02¢h- : 

Ordinary red ants (probably Monomorium latinode Mayr) are 
also possible cholera carriers, since they are omnivorous feeders 
and will penetrate to any food not carefully protected from them. 
Some experiments were conducted on these insects, but owing 
to the small volume of their feces it was difficult to make a test 
before the feces became dry. The ants ate cholera cultures and 
human feces containing cholera vibrios readily, and in one group 
of culture-fed ants cholera vibrios were recovered from the 
crushed bodies of the insects about eight hours after feeding. 
It is very probable that their feeces also contained living vibrios, 
since carmine-tinted liquid feces were discharged, but these 
became dry before they could be tested. In a second group of 
ants fed with human feces, cholera vibrios were recovered from 
the crushed bodies nearly nine hours after feeding. 

In summary, cockroaches which have fed on human cholera 
feeces may harbor cholera vibrios in their intestines, and these 
may appear in enormous numbers in the insects’ feeces for at 
least two days after the insects have fed, and may occur in 
smaller numbers seventy-nine hours after ingestion. By means 
of both feces and vomit, cockroaches may act as carriers of chol- 
era to human food. Cholera vibrios in cockroach feces will sur- 
vive on human food at least sixteen hours after discharge from 
the insect, and cholera vibrios in human feces will survive, in 
competition with numerous other bacteria, on food at least four 
days. There is no loss of virulence for guinea pigs in cholera 
vibrios after twenty-nine hours in the intestine of the cockroach. 
Cholera vibrios may be found in the bodies of ants at least eight 
hours after they have ingested cholera cultures or human feeces 
from cholera patients. 


REAPPEARANCE OF PLAGUE IN THE PHILIPPINES AFTER AN 
ABSENCE OF SIX YEARS } 


BRIEF DESCRIPTION OF THE OUTBREAK, THE METHODS USED TO COMBAT IT, 
AND THE PROBABLE FACTORS IN ITS INTRODUCTION 


By Victor G. HEISER 
(Director of Health for the Philippine Islands) 


One map 


The Far Eastern Association of Tropical Medicine has been 
a beneficent influence in disseminating useful sanitary informa- 
tion among medical men of the Orient, and has been a great factor 
in promoting friendly relationship between the countries from 
which its membership is drawn. 

In choosing plague as a subject, we deal with a disease that 
concerns all of us, because it is constantly present in some ports 
of the Orient and is one that constantly threatens our frontiers. 
The factors concerned in the transmission of the disease are now 
so generally accepted that a study of the methods of combating 
plague in the different countries shows them to be very much the 
same. 

Therefore, the purpose of this paper will be, in addition to a 
brief description of the outbreak in the Philippine Islands, to 
refer particularly to those features which have not been so 
frequently mentioned in the literature of the disease. 

After an absence of six years in human beings and five years in 
rats, plague was again found in the Philippine Islands in man 
on June 17, 1912. From the beginning of the outbreak up to 
the present time, there have been in Manila 68 cases with 58 
deaths, which gives a mortality of 85.3 per cent. At Iloilo, there 
were 9 cases with 9 deaths. In view of the fact that Manila is 
a city with a population of approximately 300,000 and is largely 
built up with a poor type of wooden buildings, which furnish 
ideal harboring places for rats, it would not have been strange 
if there had been more cases. In fact, a much larger number 
of cases could have been reasonably expected. On account of the 
almost daily communication which Manila has with plague- 
infected foreign ports which are within a few days’ steaming 
distance for the average vessel and since passengers, crew, ro- 
dents, and vermin may arrive well within the incubation period 
of the disease, it is remarkable that the Philippines should have 
remained free from plague for so many years. This freedom 


*Read at the third annual meeting of The Far Eastern Association of 
Tropical Medicine held at Saigon, November, 1913. 
5 


6 The Philippine Journal of Science 1914 


can probably be attributed to the fact that all such vessels are 
fumigated at intervals of not greater than six months with sul- 
phur dioxide and that they unload either into lighters in the bay 
or on to rat-proof wharves. 


UNUSUAL CHARACTER OF PLAGUE AT QUARANTINE 


It is perhaps worthy of note that, prior to the appearance of 
plague in Manila a number of cases of the disease was found 
on incoming steamers. For instance, on April 6, 1912, a death 
was reported on the steamship Zafiro, which had arrived the 
day previous from Hongkong and had been in the harbor for 
twenty-four hours at the time of the death. At the medical 
inspection of the vessel, which was made the day previous, no 
illness was detected. An investigation showed that the victim 
had been on deck on the night of April 5, 1912, in apparently 
good health. The next morning, at 6 o’clock, he was found dead 
in his bunk. The necropsy and subsequent biological findings 
reported by Dr. R. P. Strong of the Bureau of Science showed 
that death was due to pneumonic plague. 

On April 7, 1912, the steamer Loongsang arrived in Manila 
from Hongkong, and the captain reported that a death had oc- 
curred the day previous in a Chinese member of the crew. Upon 
investigation of this case, the captain stated that the man was 
apparently in good health, but that while hauling on a rope he 
fell over in an apparent faint and was placed in a chair and in 
the course of a few hours expired. The necropsy and animal 
inoculations showed that he had died of plague and probably 
of the pneumonic variety. 

Beginning April 7, 1912, the temperature of all members of 
the crew and of the passengers that arrived in vessels from 
foreign ports was taken with a view to detecting any possible 
cases of plague. 

On the arrival of the steamship Taisang from Amoy at the 
Mariveles Quarantine Station at about 6.30 a. m. on April 30, 
1912, the entire personnel was carefully examined and found 
free from sickness of a suspicious nature and from elevations of 
temperature. Seventy-three persons were detained to serve a 
quarantine detention of seven days. On the evening of April 
30, a Chinese passenger, aged 51 years, was found to have a 
temperature of 39° C. with a pulse of 100. He was placed in 
the hospital, but protested vehemently that he was not sick. He 
was carefully watched from the first; there was a slight cough; 
physical examination of the chest revealed a few rales; smears 
made of the sputum and stained for plague bacilli were negative. 


1X, Bot Heiser: Plague in the Philippines 7 


On the fifth day, the fever still persisted, but the patient stated 
that he did not feel ill and demanded to be released from the 
hospital. On this day, the expectoration was bloodstained, but 
no suspicious organisms could be found in the smears nor could 
any physical signs of pneumonia be detected. Furthermore, 
there were no palpable glands. On the morning of the seventh 
day, the temperature and pulse dropped and the general condition 
was distinctly worse. The patient now admitted that he felt ill. 
Several hours later, he flinched when pressure was made in the 
right axilla. Lymphatic enlargement was now made out, and 
by the evening of the seventh day the bubo in the axilla had 
increased markedly in size, the swelling approximating 3 by 7 
centimeters. Glands now became palpable in other portions of 
the body, particularly in the cervical region, and a few hours 
later there were inguinal and femoral buboes. The patient 
became rapidly worse, and died at 7 o’clock on the morning of 
the eighth day of his illness. At the necropsy, the glands of the 
right axilla and those of the right side of the neck were found 
enlarged; the other lymphatic glands were also enlarged, but to a 
lesser degree. There was consolidation of the lower lobe of 
the right lung, and the spleen was about twice its normal size. 
In brief, the necropsy findings of a typical case of septiczemic 
plague were present. Smears from the spleen and the right 
axillary gland showed immense numbers of bipolar-staining 
organisms. Cultures made from fresh pieces of tissues and 
later inoculated into animals gave positive results for plague. 


ROUTINE RAT PRECAUTIONS PRIOR TO HUMAN OUTBREAK 


The city of Manila is divided into 5 sanitary divisions. Each 
division is in charge of a medical officer, who has a corps of from 
10 to 40 sanitary inspectors to assist him. During the entire six 
years during which plague was absent, test rat catching was 
done at weekly intervals under the direction of each station at 
places that were believed to be liable to become rat-plague in- 
fected, and in addition any unexplained mortality among rats 
was always investigated, but at no time were plague rats found. 


HUMAN CASES 


In view of the.foregoing, it was with considerable surprise 
that the first case of plague was found in a human being. This 
case was discovered June 17, 1912, in a Filipino employed as a 
watchman at 236 Calle San Jacinto, which is in the Chinese 
district, his residence being at 920 Calle Antonio Rivera, which 
is the slum section of the city. 


8 The Philippine Journal of Science 1914 


The victim was found dead at his home with a history of 
having been ill for about three days. On post-mortem exami- 
nation, typical plague buboes were found in the right groin and 
axilla. Inoculation into guinea pigs resulted in typical attacks 
of plague. The victim was a permanent resident of Manila, and 
had not been away from the city in many months. He worked 
and lived in a section that is far removed from the water front, 
and did not associate with persons who had been out of the city 
or persons who were connected with shipping. So far as known, 
the nearest focus of the disease was Hongkong. Therefore, 
the source of this infection was difficult to explain. 

The next case was found June 26 in a Filipina woman who 
lived at 1615 Calle Azcarraga near the Arranque market, which 
is over a kilometer from the place where the previous victim 
either worked or lived. 

The next case did not occur until August,4, which was thirty- 
nine days later. - This victim resided at 139 Calle Villalobos in 
Quiapo, which is a large retail district located near the Pasig 
River and fully 1.5 kilometers from where the previous case had 
occurred. 

The next case occurred on August 7 on the same street and 
block as the preceding case. 

The next case occurred on August 21 at 352 Calle Echague, a 
street corner, which is also in the same block as Calle Villalobos. 

It may be of interest to note that these last three cases were 
all in schoolboys under 16 years of age. 

By October 20, there had been 13 cases from the beginning of 
the outbreak, and these occurred at irregular intervals and in 
different sections of the city. Then between the dates of October 
20 and October 22, 13 new cases occurred, so that in a period 
of two days there were as many cases as there had been during 
the four preceding months. 


EXPLOSIVE HUMAN OUTBREAK 


An investigation showed that in these latter 13 cases there was 
definite geographic grouping, which was a feature that had not 
occurred up to that time. The victims were all laborers who 
worked at the freight station of the Manila Railroad Company. 
Inquiry developed the fact that large numbers of rats had been 
seen dying first in the north warehouse and a few weeks later 
in the south warehouse. About three weeks after the heavy 
rat mortality was noticed in the north warehouse, plague ap- 
peared among the laborers in the south warehouse. The ware- 


1X, B,1 Heiser: Plague in the Philippines 9 


houses (godowns) were galvanized iron buildings with a dirt 
floor, on sections of which a board floor had been constructed. 
Numerous rat runs were found that led under the boards, and 
upon these floors being removed rat nests and a number of dead 
rats were found. The rats were mummified, rendering it im- 
practicable definitely to ascertain the cause of their death. It is 
customary in the Philippines for laborers to go barefooted and 
barelegged as far as the knee, and as these employees were no 
exception there was ample opportunity for fleas to bite them on 
their lower extremities. 

From October 22 until October 31 there were 8 additional cases 
traceable to the warehouse, and then the outbreak stopped as 
suddenly as it began. The preliminary sanitary measures were 
completed by October 25, and as there were no further cases after 
the incubation period had expired it seems fair to assume that 
the sanitary measures were completely effective. 

The control of this explosive outbreak probably furnished as 
good an example of the effectiveness of modern sanitary measures 
against plague as can be found in the literature of the disease. 
As there were several hundred laborers employed in these freight 
warehouses, there is every reason to believe that large numbers 
of additional cases would have occurred if the proper steps had 
not been promptly taken to eliminate the infection. 


PLAGUE AMONG RATS 


Upon the discovery of the first case of human plague on June 
17, a careful investigation was again made, but no history of 
any unusual mortality among rats could be elicited. Immediate 
steps were taken to catch rats in the sections of the city in 
which the victim had worked and where he had lived. This 


- work was very actively carried out, but it was not until August 


31 and until over 7,000 rats had been caught that a plague rat 
was detected. This rat was caught in a spring trap at 351 Calle 
San Sebastian, which is in the block in which the human case 
on Calle Villalobos had occurred during the first week in August. 
On September 7, a plague rat was found at 104 Calle Santa Rosa 
and another at 215 Calle Echague, both of which addresses are 
within a block of the case that occurred on Calle Villalobos. 
On October 4, a plague rat was found at 644 Calle Ilaya and 
another at 637 Avenida Rizal, which are sections far removed 
from where either human or rat plague had occurred heretofore. 
During the early part of October, the rat-catching efforts were 
increased and rats were caught at the rate of approximately 


10 The Philippine Journal of Science 1914 


9,000 per month, but the percentage of infected rats found up 
to November was only 0.005, which is unusually low. According 
to many authorities, a 2 per cent rat-plague infection is con- 
sidered a low average. At Hongkong, for instance, it has been 
reported that 7 per cent of the rats examined prove to be plague 
infected. 

PLAGUE AMONG OTHER ANIMALS 


On November 26, 1912, five dead rats were reported from the 
United States Army Commissary warehouse, which is located on 
the Pasig River near Malecon Drive and is on the south side of 
the Pasig River. All infections heretofore had occurred on the 
north side of the river. Unfortunately the rats were thrown 
into the river and, therefore, the causes of their death could 
not be ascertained. 

On November 27, a cat known to have caught and eaten rats 
in this Commissary warehouse was reported to be sick and was 
taken to the Bureau of Science, where it was observed for a 
period of three days, at the end of which time it died. At the 
necropsy, typical bubonic cervical plague glands were found, and 
inoculations made into guinea pigs from material from the spleen 
and buboes produced typical plague in the guinea pigs. A guinea 
pig that was inoculated by a swab introduced into the cat’s 
rectum also died from plague. 

Eighty rats were caught in this and adjacent warehouses, but 
none of them showed any evidence of plague. 

On December 17, a woman died of plague at 4 Calle Barraca. 
Two hours later, Doctor Jackson, the medical officer in charge 
of the antiplague measures in Manila, and Doctor Schobl, the 
laboratory representative of the Bureau of Science, placed 2 
healthy guinea pigs free from plague in a wire cage upon the 
petate (mat) which was located on the floor and on which the 
woman slept, and left them there for one day; the routine 
insecticidal measures were delayed until after the guinea pigs 
had been removed. On December 21, one of the guinea pigs 
died from typical bubonic plague, the diagnosis being fully con- 
firmed by inoculations into other animals. The guinea pigs were 
carefully searched for fieas, but none could be found. Guinea 
pigs under similar circumstances were placed in two other 
houses in which plague had occurred, but after the disinfecting 
and insecticidal measures had been completed. In neither case 
did plague result in the guinea pigs, which would indicate again 
that the sanitary measures employed were effective. 


IX, B, 1 Heiser: Plague in the Philippines 11 


RARENESS OF SICKNESS OR DEATH FROM PLAGUE IN RATS 


An interesting incident in connection with the plague out- 
break was the rareness with which rats. were found that were 
either sick from plague or dead as a result of plague. Among 
a total of 37 rats in which plague was detected, a rat sick 
of plague was found only once, and rats dead of plague were 
found in but three instances, two of which did not occur until 
September, 1913. Of the remaining rats that were found to 
be afflicted with plague, 20 were killed in a spring trap, 8 died 
as a result of poison, 3 were killed with a club or other weapon, 
and in 2 the cause of death was not ascertained. 


[ RAT NESTS 


The principal measures in eradicating plague were directed 
toward finding and destroying rat nests. It was thought that 
in this way the danger of infected fleas spreading the plague 
among rats could be largely eliminated, and the small number 
of cases of rat-plague infection which occurred may be explained 
through this sanitary measure.. Experience shows that fleas 
spend much time in rat nests, and by disturbing the nesting place 
and by spraying it frequently with an insecticide the number 
of opportunities for fleas to spread the disease is certainly very 
much lessened. In order to accomplish this purpose, a gang 
of laborers composed of about 100 men was divided into three 
sections, each under the charge of an experienced sanitary 
inspector. These gangs go from house to house, and make a 
thorough systematic search for rat-breeding places. After the 
search of the house is completed, the woodpiles, old junk, rub- 
bish, and everything that is liable to harbor rats outside of the 
house are moved about so that the nests may be discovered and 
destroyed. Particular attention is given to spraying with an 
insecticide so that fleas may not escape. It was found that 
woodpiles are favorite harboring places for rats, and these are 
invariably taken down and repiled well above the ground and 
away from the wall, so that dogs and cats may work effectively in 
keeping rats out of them in the future. 

In view of the great importance which has been attached by 
the Javanese sanitary authorities to destroying rats in hollow 
bamboos, special stress was directed toward finding them in 
similar breeding places in the Philippines. At first, these efforts 
did not meet with much success. However, when the plague 
spread to sections of the city in which bamboo and thatched 
houses predominate, rats were frequently found breeding in 


12 The Philippine Journal of Science 1914 


the hollow bamboos, and our experience in the Philippines there- 
fore fully confirmed the experience of the Javanese authorities. 


RELATIONSHIP OF HUMAN TO RAT PLAGUE 


In the 68 cases of human plague, rats afflicted with plague 
were only found on three occasions in the house or premises in 
which the victim died or worked. The first of these instances 
has already been mentioned under the head of explosive out- 
break at the Azcarraga station of the Manila Railroad Company. 
The second occurred in a jewelry shop, and will be discussed 
under the head of multiple house infection. The third was that 
which resulted in the death of the editor of the Manila Daily 
Bulletin. 

It is also of interest to know that human plague existed for 
over two months before any rat plague could be found. Not- 
withstanding the foregoing, an examination of the map in the 
appendix of this report shows that by districts there was a very 
close relationship between human and rat plague in Manila, and 
in none of the human cases, after rat plague had once been 
discovered, could a history be obtained that the human victim 
had not been in the district in which plague rats were found. In 
every instance, he either lived or worked in such a section. 


MULTIPLE HOUSE INFECTIONS 


The first instance of multiple house infection was that already 
described under explosive human outbreak, which occurred at the 
freight warehouse of the Azcarraga railway station. The next 
was reported on February 18 at 1028 Calle Comercio. A Chinese 
boy, aged 15, died of plague on the 12th, and a Chinese, aged 50, 
died of plague on February 18. During the week ending April 
27, there were 5 cases of plague, all in Filipinos, that occurred 
among the employees of a silversmith shop at 1364 Calle Sande. 
These men were all employed on the first floor, which was of 
cement. Ordinarily, such construction would be regarded as rat 
proof; but, owing to the openings which resembled rat holes, 
found near the sides, and cracks located here and there, it was 
deemed advisable to tear out the floor. Several mummified rats 
were found underneath, the death of which was in all prob- 
ability due to plague, although this fact could not be definitely 
established. It is assumed that the fleas made their way through 
the cracks in the floor or ‘perhaps left a plague-sick rat during 
its migrations through the shop. This was another striking 
instance of the close relationship which exists between rat and 
human plague. 


— * 


ee 


IX, B, 1 Heiser: Plague in the Philippines 13 


On May 15, there was a second death from plague at 1226 
Calle Juan Luna, the first cases having occurred on May 14. 

A review of the history of the plague outbreak in Manila 
shows that the disinfecting and insecticidal measures taken at 
each house must have been effective, because there is not a 
single instance on record of a second case of plague having oc- 
curred in any house after the incubation period of the disease 
had expired. 

SEASONAL PREVALENCE 


The following table shows the prevalence of plague by months. 
The seasonal prevalence of plague in many Oriental ports is of 
the greatest epidemiological significance. In Hongkong, for in- 
stance, the greatest number of cases for many years has gen- 
erally occurred in May, and the smallest, in December. In Amoy, 
which is several hundred kilometers farther north, the greatest 
number of human cases occurs in June. The following table 
indicates that the greatest number of cases occurs in the Phil- 
ippines in October, but this can be considered more or less ac- 
cidental, because the majority of those that occurred in October 
were due to the explosive outbreak which was traced to the 
Azcarraga railway station, and that can scarcely be ascribed to 
seasonal reasons. There is also placed alongside the human cases 
the incidence of rat plague, which shows that the greatest number 
of plague rats was found in March. A study of these would 
seem to show that if the often-quoted observation is true that 
human cases should follow two months after rat plague has 
occurred, the greatest number of human cases should have oc- 
curred in Manila in May. The sanitary measures which were 
so actively carried out against rats during March and April may 
have prevented this. 


TABLE I.— Distribution by months of human and rat plague cases in Manila. 


= Human Rat | Human octet 
Month and year. plague | plague | Month and year. | plague | plague 
cases. | cases. | cases. | cases. 
Trees | | ; 
1912. 1913. | | 
Suing er SLE A: hes 2 Ojphebruary=-6=5- 42555. -20.. 3 | 1 
GN LC) recs ec ee 0 Uy) RTH oes ee se 4 | 13 
PRISE eee oe ee oe ee 5 Tod | / G7) ort ae es 2s ee 6 | 3 
September = =.2- 6 25. ose 8 BN ety ete eae uae oe aes Le 3 | 7 
October i225 eek 21 Td | Ares Cet See i oy es a oe 0} 3 
November ic..-- Say. fe du! 2 (| Pa Tic A & e Sees SOE eee 0 | 0 
December 4.5 eek ee ee 6 3 | (Nata Re a ee 1 | 0 
1913. ikGeptember’.- 22.2. .-2 22.2 1 3 
January Cot ee nee 1} 0 || October ---.-----------__-_-- | 0 0 


14 The Philippine Journal of Science 1914 


EVIDENCE OF FLEA TRANSMISSION OF PLAGUE 


On account of the completeness of the results obtained by the 
investigation of the death from plague of William Crozier, the 
editor of the Manila Daily Bulletin, it is thought that it might 
be of interest to give a brief description of the findings. Mr. 
Crozier felt ill on the evening of September 18, 1913. On the 
morning of the 19th, he was admitted to a local general hospital. 
By afternoon he developed symptoms suspicious of plague, and 
microscopical examinations made on September 20 showed typ- 
ical bipolar-staining organisms. He was immediately removed 
to the San Lazaro plague hospital, where he died on September 
22 from bubonic plague. The diagnosis was biologically con- 
firmed. On September 6, a plague rat was found in the block 
next to the one in which the building in which he worked was 
located. A mummified plague rat was found in one of the 
drawers of Mr. Crozier’s desk in his office. A number of fleas 
were seen hopping about, and one of these was captured and 
definitely identified at the Bureau of Science as a specimen of 
Pulex cheopis. Upon the flea being ground up and stained 
specimens made, a bipolar-staining organism was found. In the 
meantime, the mummified rat that was found in the desk was 
also ground up, and inoculations made into healthy laboratory 
rats resulted in typical cases of plague. This point is interesting 
for a number of reasons. In the first place, it shows that live 
fleas may harbor virulent plague bacilli for a period of at least 
two weeks, because it can be stated with certainty that the rat 
must have been dead for at least that period of time and probably 
very much longer. The possibility, of course, remains that the 
fleas might have come from rats that visited the drawer after 
the rat which was found had died. If, however, the fleas were 
from the dead rat which was found, it shows that fleas may 
live at least two weeks and harbor plague bacilli during that 
period. It does not seem probable that the fleas fed on the 
dead rat and thus ingested plague organisms after the rat’s 
death. 

In view of the foregoing findings it would appear possible that 
plague might be introduced into a country by infected fleas. 


SANITARY MEASURES EMPLOYED 


On account of the district sanitary organization which exists 
in Manila, no particular additional organization was required to 
combat the plague, except to employ a force of laborers for the 
purpose of catching rats and carrying out general cleaning-up 


BAe Be _ Heiser: Plague in the Philippines 15 


measures. The medical officer in charge of a district in which 
plague occurred ordered the immediate transfer of the case to 
the San Lazaro plague hospital and called upon the disinfecting 
squad to spray or wet down with kerosene the premises in which 
the cases occurred. This was later followed by disinfection 
with larvicide, a preparation which is used in Panama as a disin- 
fectant and as a larvicide. It makes a milky solution upon being 
added to water. It is serviceable both as an insecticide and as a 
disinfectant, particularly where greasy surfaces have to be dealt 
with. 

When a case of human or rat plague was found in a house, 
the house was regarded as a plague center and the infected area 
was arbitrarily considered to be three blocks on each side of it. 
Rat-catching operations were begun on the periphery of this 
zone and gradually directed inward until the infected house was 
reached. It was thought by proceeding in this manner that 
there was less danger of driving infected rats to other portions 
of the city and also that it gave the best hope of eradicating 
the rat infection. If an active campaign against rats in an 
infected house is begun, there is great danger of driving the 
rats away from it, or, in other words, of driving rats before 
the sanitary squad and thus extending the area of the rat-plague 
infection. 

Rats were killed by means of traps and poisoned bait and by 
dogs and with clubs at the periphery of the zone. As the rat 
catchers gradually moved inward, they were immediately fol- 
lowed by from 50 to 100 laborers for general cleaning operations. 
Each house was entered. Barrels, boxes, furniture, piles of 
mattresses, bedding, straw, or any other things among which 
rats might hide were moved about and replaced in such a manner 
as to insure that such places were free of rats. The premises 
connected with the house were then treated in a similar manner. 
Rubbish, straw, old boxes, and other similar articles were sent 
away and burned at the central crematory. Wood-piles, boxes, 
and other articles which could not be treated in this way were 
taken down and neatly repiled, well above the ground and free 
from the wall, so that rats would be accessible. While these 
operations were going on, specially trained fox terriers were 
kept on guard, and as the rats attempted to escape from their 
hiding places they were caught by the dogs or clubbed to death 
by the laborers. Large numbers of rats, and particularly rat 
nests, were destroyed in this manner. As soon as these opera- 
tions were completed, rat-proofing measures, so far as practi- 


16 The Philippine Journal. of Science a 


cable, were carried out; rat runs were obliterated; where neces- 
sary, ground surfaces were cemented; and all harboring places 
- were destroyed wherever practicable. By the time that these 
operations had extended to the infected house, the sanitary 
engineer of the Bureau of Health made a careful inspection of 
the infected house and issued the necessary sanitary orders to 
make it rat proof. In accordance with the Manila ordinance, 
the Bureau of Health has authority to order even extensive 
structural changes in houses in which plague has occurred. 

In the meantime, the sanitary force connected with the station 
of the district was busily engaged in making house-to-house 
inspections in order to ascertain whether or not there were any 
additional human cases. Constant inquiry was made as to 
whether any unusual mortality was apparent among rats. The 
public was particularly requested to report all dead rats found, 
and these were promptly taken to the laboratory. 

Active steps were taken to deprive rats of their food supply. 
All garbage was put into metal containers covered with tight- 
fitting lids. Hay, oats, corn, fodder, and feed for animals gen- 
erally were ordered placed in rat-proof containers. 

More extensive rat catching was immediately begun in sus- 
pected areas in all other districts of Manila in order to ascertain 
whether there was any other rat-plague infection. 

It was hoped that by employing such means, accurate infor- 
mation would be available for detecting an outbreak in advance 
and taking sanitary measures before human cases could occur. 


PREVENTION OF SPREAD OF PLAGUE FROM MANILA TO THE 
PROVINCES 


At the railway warehouses, men were stationed to inspect all 
cargo that was shipped from the city, and in many instances 
in which it was suspected of containing rats it was repacked. 
To prevent the spread of the disease by sea, all vessels were 
required to use rat guards, and fumigations were made at in- 
tervals of a few months. All lighters, cascos, and other craft 
used in transporting cargo from ships to shore were also included 
in these fumigations. All ports in the Philippines to which these 
vessels proceeded imposed antirat regulations against such vessels. 
They were required to use rat guards wherever they went along- 
side of docks or piers and also to undergo medical examination. 
Fortunately, the water front of Manila and the warehouses from 
which ships load did not become rat-plague infected. 


CC Se 


ESB, 1 Heiser: Plague in the Philippines Ay 


FUMIGATION OF VESSELS 


In the appendix will be found complete tables, by years, show- 
ing the number of vessels fumigated with sulphur in Manila 
since 1903 and the number of rats found on them. All of the 
foregoing data have been made into a consolidated table, and 
it is of interest to note that there is little difference between 
the number of rats killed per vessel as shown by the consolidated 
table as compared with the table for any one year, which is 
strong evidence that vessels must be fumigated at frequent inter- 
vals, because vessels that have been regularly fumigated are just 
as liable to harbor rats as vessels that have not been fumigated. 


EFFECTIVENESS OF THE VARIOUS MEASURES EMPLOYED IN 
CATCHING RATS 

The ratio maintained in catching rats with two types of traps 
is well shown in the following table, a perusal of which will 
show that for the three months ending June 30, 1913, there 
were 120,565 spring or snap traps set and that for every 100 
of this type of trap set there were caught 6.9 rats. During the 
same period, there were 47,075 wire-cage traps set; the total * 
number of rats caught was 339, which gives 0.72 rat caught for 
each hundred traps set. For the quarter ending September 13, 
130,627 spring or snap traps were set and 9,753 rats caught, 
which gives 7.47 for each 100 traps set. During this period, 
40,621 wire-cage traps were set and 395 rats were caught, which 
gives 0.97 rat caught for each 100 wire-cage traps set. 


TABLE II.—Relative efficiency of poisons and different kinds of traps used. 


Quarter ending June 30. Quarter ending September 30. 
. . Number Number 
Kind of trap or poison. 
Number gaa Percent- | Number oe Percent- 
set. or poi- age. set. or poi- age. 
soned. soned, 
Spring or snap traps ________-______- 120, 565 8,377 6.900 | 180,627 7, 753 7.47 
Wire-care trapae----- 22-2 47, 075 339 0. 720 40, 621 395 0.97 
Poisoned bait: 
Bacon and coconuts_-____-_--___- 166, 237 1,216 CU GHN | tele See oles oe 2 ee nee 
Goconata, a lee ce eee ae oe aA ee obs Secale eke 177, 309 216 0.12 
Number of rats caught by dogs__-_.--_-----_------------ 160 [S| Sent See a ee 
Number of rats killed by clubs and other weapons-__--_---- 2, 889 SERt Bil 2 ee Oe Se 
Number of rats found dead from other causes ___-_--_--- 316 DONT ieee Oa 5 cata 


No accurate account was kept of the various forms of rat 
poison used. Bacon, coconut, and rice were used in different 
1242932 


18 The Philippine Journal of Science 1914 


formulas. For instance, for the quarter ending June 30, 1913, 
there were 166,237 poisoned baits set and the rats found poisoned 
averaged for each 100 baits 0.72, from which it appears that 
the rat poison ranks lowest in efficiency, but perhaps highest 
in economy. In view of the fact that the original cost of the 
cage trap is many times that of the spring trap and that 
the cost of maintenance is very high, it will be apparent that 
the spring trap is by far the more economical as well as more 
effective of the two. 


MANILA RAT-PROOFING REGULATIONS 


On account of the presence of plague in Manila, it was deemed 
most opportune to insist not only upon rat proofing in the 
buildings and areas which were infected, but to require rat- 
proof construction in all buildings which were to be erected 
in Manila in the future, and the following regulations have 
now been enforced during the past six months. 

All proposed new buildings of whatever nature, whether fac- 
tory, stable, garage, bodega, warehouse, private dwelling, or any 
other class of building, shall be designed and erected so as to 
have no hollow ceilings, walls, columns, stairs, floors, ete. This 
shall also apply to repairs or alterations to existing structures. 
Whenever a ceiling or hollow partition is removed, the same shall 
not be replaced. 

Architects, builders, and others concerned are requested to 
incorporate the following paragraphs in plans and specifications 
for future buildings of this nature. 

For the purpose of preventing the entrance and harboring of 
rats etc., this building will be constructed (or repaired) without 
hollow walls, hollow ceilings, hollow stairs, hollow floors, hollow 
columns, etc. 

All walls, with the exception of solid wood framing, within 
1 meter of the ground will be of concrete, brick, stone, mortar, 
or other material proof against the incursions of rats and will 
extend below the ground to a depth of at least twice the thick- 
ness of the wall. 


KINDS OF RATS CAUGHT IN MANILA 


From the beginning of the outbreak until September 30 there 
were 68,667 rats caught. At the outset, an effort was made to 
classify the different species of rats, but owing to the difference 
of opinion among those charged with the work as to the correct 
identification of many of the rats it is not deemed that the figures 


E,B,1 Heiser: Plague in the Philippines 19 


are sufficiently reliable to quote them. However, since August 
13 to September 27, 1913, rats have been classified as follows: 
Gray rats, 1,103; black rats, 220; shrews, 821; unclassified, 
981. 
PLAGUE IN ILOILO 


In Iloilo, a case suspicious of plague was reported on July 5, 
1912, and this diagnosis was subsequently confirmed by the 
laboratory. It occurred in the person of a Chinaman who was 
reported to have come from Bais, Oriental Negros, but later 
investigation showed that he had been a resident of Iloilo at 
least since February, 1912. The next case was reported Au- 
gust 18, and the last case, September 17, 1912. There was a 
total of 9 cases. All of the cases were confined to two houses. 
During July, August, September, and October, 1,146 rats were 
caught in the vicinity of the houses in which the human cases 
had occurred, along the water front, and in the places which were 
regarded as suspicious, but in not a single instance was an 
infected rat found. 

Doctor Fox, who was in charge of the antiplague measures, 
concluded that there was a possibility that the disease might have 
been imported into Iloilo, either from China or from Manila, by 
means of bedbugs. In view of the experience had in the death 
of Mr. Crozier, which occurred on September 22, 1913, it 
would also appear possible that plague-infected fleas might have 
been introduced into Iloilo and they might have been responsible 
for the outbreak. However, there is no scientific proof, and the 
actual facts as they occurred are only stated for what they are 
worth. 

PLAGUE NOT A FILTH DISEASE 


Our experience in the Philippines shows that plague is not a 
filth disease. A well-to-do citizen is as liable as the slum dweller 
to become infected if rats infect his house or other places that 
he frequents. In Manila, some of the worst slum sections of the 
city escaped; on the other hand, some of the better sections in 
which there were large stores of food which attract rats became 
infected. 


PROBABLE FACTORS CONCERNED IN THE INTRODUCTION OF PLAGUE 
INTO THE PHILIPPINES 


Much time was spent in collecting data which, it was hoped, 
would show in a scientific manner how plague was introduced in 
the Philippine Islands. In addition to the routine rat catching 
which has been practiced for more than ten years, immediately 


20 The Philippine Journal of Science 1914 


after the disease made its appearance in Manila, large numbers 
of rats were caught along the water front and near the wharves, 
but none of them were found to be plague infected. The rat- 
proof piers at which foreign vessels lie have been all that their 
description implies; they have remained rat free. On account 
of the fact that the first cases occurred among permanent resi- 
dents and among persons who had not been out of Manila in many 
months and who did not associate with people who worked along 
the water front or with persons who had recently been in a 
plague-infected country, it seems reasonable to infer that the 
disease was not introduced by human beings. As no infection 
could be found among the rats of the water front and especially 
since the wharves remained free from rats, it does not seem 
probable that infected rats could have come from a ship by means 
of gangways, cargo chutes, lines, or by other direct means. All 
vessels that ply in Philippine waters are fumigated at least 
twice annually; vessels from ports that are suspected of being 
infected with plague are fumigated every other trip; and vessels 
that carry rice or other food supplies which are especially liable 
to carry rats are fumigated every trip; so that the liability of 
plague rats coming ashore directly from ships is extremely im- 
probable, and moreover none of the rats found in such ships in 
the course of the fumigation work showed any evidence of plague. 

There does remain, however, the very strong probability that 
plague rats may have been introduced into Manila in cargo and 
may have made their escape therefrom after it was delivered in 
the city. This is possible in view of the enormous quantities of 
food supplies and other cargo that come directly from plague- 
infected centers in China and Japan. For instance, every week 
there are literally thousands of baskets or crates of eggs, garlic, 
onions, and similar foodstuffs, among which rats can easily 
take refuge, that come from places like Canton and Amoy within 
a period of five days. It is well known that plague has existed 
in Canton, for instance, almost continuously during the past ten 
years, and it is not improbable that plague rats might have been 
introduced into the Philippines from that port. Large quantities 
of cargo arrive from Japan as, for instance, glass- and china- 
ware, bottles, and other things which are packed in hay and 
straw. Among these a rat might easily have been brought into 
the Islands. 

In a case of human plague which occurred at 508 Calle Mag- 
dalena, bedbugs were found on the petate upon which the man 


1X, B, 1 Heiser: Plague in the Philippines VA 


died, and smears from the intestinal contents showed plague-like 
bacilli. From the foregoing, it would appear that perhaps in- 
fected bedbugs might be concerned in spreading the disease. 

The recent experience already described in this paper, in which 
live fleas containing plague bacilli were found in the desk of Mr. 
Crozier after the rat must have been dead for at least two weeks, 
would also seem to make it possible that plague might have been 
introduced by infected fleas, although the liability of such in- 
troduction is greatly reduced because the clothing and other 
effects of all second- and third-class passengers are disinfected 
with steam. 


CONSIDERATIONS WITH REGARD TO A PRACTICAL METHOD TO 
PREVENT THE INTRODUCTION OF RATS OR INSECTS IN 
CARGO OR PASSENGERS’ EFFECTS 


Since attention has been directed to the possibility of rats 
being present in cargo, it has been a frequent experience for 
customs employees and warehousemen to report rats in cargo 
during unpacking operations. For instance, rats are found more 
frequently among onions than among potatoes. Certain varieties 
of potatoes are much more popular with rats than others. Rats 
or insects that are free in the holds or other places on a ship 
can be eradicated fairly successfully by fumigation while the 
cargo is in course of being discharged or even afterward. 

Experience has shown that rats that are actually concealed 
in cargo cannot be reached with sulphur gas with any degree of 
certainty while such cargo is in the hold. There is also the 
important consideration that when a gas is used which is suf- 
ficiently strong to penetrate the containers, there is great danger 
of injuring cargo like cloth, camphor, tea, etc. An effort was 
made to unpack all cargo liable to harbor plague rats in a rat- 
proof room or inclosure, but the cost was found to be prohibitive. 
A conference was held with the importers for the purpose of 
having an expression of opinion as to whether cargo liable to 
harbor rats could not be shipped in metal containers, but these 
last two procedures were declared by them to be too expensive 
from a commercial standpoint. 

A review of the literature of plague fails to reveal any plan 
effectually to deal with the prevention of the introduction of 
rats in cargo. It is obvious that this is a most important con- 
sideration, especially when cargo can be delivered within ten days 
from the time that it was shipped from an infected port. 


2 The Philippine Journal of Science 1914 


SUMMARY 


After an absence of six years in human beings and five years 
in rats, a case of human plague was found in the Philippines 
on June 17, 1912. There have been a total of 68 cases and 58 
deaths up to October 1, 1918, in Manila, and 9 cases and 9 deaths 
in Iloilo. During April, 1918, several cases of pneumonic plague 
were detected on vessels that came from Hongkong and Amoy. 
A careful investigation of these cases and of all subsequent 
arrivals failed to show any connection between them and the 
first cases of plague on June 17 in Manila. The disease was 
probably introduced by plague rats or insects present in cargo 
from infected ports which was not unpacked until it was distrib- 
uted in the city. Rat catching was done in Manila during the 
entire time that plague was absent, but no case of rat plague 
was found until August 31, 1912, this in spite of the fact that 
over 14,000 rats had been caught in districts in which human 


cases had occurred since June 17. Plague was found in rats. 


and cats and in bedbugs and fleas. An explosive human outbreak 
occurred in October in which 21 cases were traced to the goods 
warehouse at the Azcarraga railway station. The gray rats 
were found to be the commonest. The percentage of plague 


among rats has been very small, less than 0.002, whereas it is © 


the common experience in cities in which plague occurs that at 
least 2 per cent of the rats are plague infected. Another most 
striking incident was the fact that of the total 48 plague rats 
which were encountered, a rat sick of plague was only found once 
and a rat that died of plague was only found once. The remain- 
ing rats in which plague was detected were caught in spring 
traps, died as a result of poison, or were clubbed to death. The 
transmission of plague by fleas was definitely shown by guinea 
pigs contracting plague from the bed of a human victim and by 
finding infected fleas in the desk of a human victim. Multiple 
house infection occurred only three times, and all of the cases 
were within the incubation period of the disease. Rat nests 
were frequently found in hollow bamboo, and the experience 
had in Java was fully confirmed. Seasons apparently had no 
influence upon the number of cases, whereas in the near-by ports 
of Hongkong and Amoy seasonal prevalence is most marked. 
The only place in the Philippines in which plague occurred outside 
of Manila was Iloilo. The sanitary measures employed consisted 
in the isolation of the plague victim in a plague hospital. The 
rat-catching and rat-proofing measures were begun at the pe- 
riphery of a zone which extended three blocks on each side of 


ee 


1B; Heiser: Plague in the Philippines 93 


the house in which the plague infection had occurred, and this 
was apparently successful in preventing extensive spread of 
plague among rats. All new buildings now erected in Manila 
or extensive repairs in old ones must be made rat proof. A 
special set of building regulations for this purpose has been 
promulgated. Not much stress was laid upon disinfection. The 
principal reliance was had upon using petroleum as an insecticide. 
The rat-catching measures were supplemented by a general clean- 
ing up in a suspected zone of all premises by a force of several 
hundred laborers, which resulted in discovering the hiding places 
of many rats and the destruction of rat nests. Particular atten- 
tion was given to finding rat nests and destroying them, with 
the hope that in this way the transfer of plague-infected fleas 
to healthy rats would be best obviated. Dogs were extensively 
employed in these cleaning-up measures, and they caught many 
rats during the process. Manila’s experience indicates that 
plague is not a filth disease. The spread of plague to the prov- 
inces was prevented by the inspection of all cargo that was sent 
out in order to be certain that it harbored no rats. From careful 
statistics kept with regard to the effectiveness of various forms 
of rat traps it was ascertained that the spring or snap is ten 
times as effective as the wire-cage trap and that the wire-cage 
trap is more effective than one portion of poison. 


1914 


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‘anbojd fo sasp)— JI] AIAVL 


25 


Heiser: Plague in the Philippines 


IX, B,1 


26 “TBI 
TZ “1B 
9 “IB 
8ST “Ww 
8ST “Fn 
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aos aaa O1dIEUIOD EZ0T |-------~--- eUON |--- afewa,, |-----eseuryD 
Snir ae saa pecs B[INBY TZT |-------~ guepnyg |----- op=---|---- ourdy ny 
ae ee ae OLOWIOY Op B[S] 96 |----~--- Jo1oqe'yT |------a]eyy |-~---eseuryoD 
Ser eee Ree oOpuoulg op o1e4sqy 78Z | ---~- ~~" @UONY |--- ~~ Opa ---|--- ~~~ Op-=--- 
eae eae ae OROLTS 1 2 FRESE 0) gos EOS 0) oa SET (0) oe 
Seer OPI!d8D 782 |---~~ eFtWesnoy |--- opeureg |---- eurdy nT 
i... a PIIPB 22g |------~- FUBAIag [~~~ op-——-|---- oud 
fone ens OzuO,Y BIOpoay, 8Tg |--~------~ op-----|----- op----|-----eseuryo 
poms eee ee ee seplavueg Z16 |-------- toaxoqey |------ereyy |---~ ourdyrg 
Sees ee sorou0qer 90g |---~ aoyevuIsserg |--- efeute,g |---- eurditg 
SSSSSce=S enseo[Uy ‘1o1te,Ul eggy |-------- aeatoqery |----- op----|------ op----- 
SSS ea See Opiiqeg 062 |------_____ eu0N |----- op-—--|-=--—- op---—= 
poses opueuseag weg 7g |---------- aopey, |----- op----|------ op----- 
SSR acl opueuse,g weg gog |----------- euoyy |----- op---~|------ op----- 
poe ae sea Seens uedn3eq gz[t |-------- aeaoqery |----- op----|------ op----- 
Rego sages caret a elulouoom 90g |-------- zeyedeo |------oyeq |---- ourdiy 1g 
Seog eae ay opususe,, weg 90g |-----sseaqsuveg |---eyeuro,g |---~ eurdiig 
PEar ey ee OuB[Og [e10Ua5 gTy, |------a0Suesseyy |----- op----|------ op----- 
: -a0yueidde 

peas iy aa opueuieg UES 70g | YFLUISTOA[IGg |----- op----|------ op----- 
a oe mae Eee InSejLW ESZl |--~~--- 1090a][09 |----- op----|------ op----- 
an ig a BIOAIY OLU0JUY [16 |------~ JopeBreg |----- op----|------ op----- 
Rapes Seer ae (eWOU0o) OP gil saree =a OPaa sea leooo > (OP aco 5 | cooaearOpaaces 
BERG Seer exeleozy 97g |--~---~~ Jeaoqe’] |----- op----|---- ourdyng 
Seer ee era oe 891BZeAe'T gzg |------£oq esnoy |----- op----|-----aseuryo 
peerage Ta OWED ZOR lana cs cae ODia casas e 0 Prare| soca Opa ooo 

powers OPSsss2|s- 2 = Ore calssa a ORs ass 


aera enseo[Uy “101187! T18T 


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wenn nn nnn en nnn nn nnn enna nn nnn nnn 80}UBS X97 | 99 
wan 2-2 = +22 = 5-2 == + =~ ----- Beqiseyl ueqojsy |; Gg 
+ aee-------------------------- BIOWIBZ OPIpPUD | FG 
aan n ana n ne tenn nasa nn nn anne nnn enna ey BnyO | es 
a----------------------------------- uBy snyyO | 2g 
wean nn nnn nnn naan ooreyl O1peg | TS 
------------~-----~------------ Sueg suop og | 09 
i Wry Bivqieg | 67 
a2 2---------------------- ejueiney Bipuelely | gp 
an 2----------------------- IZA [Op UoLoeoyiaing | bP 
------------------------ eJUOUIOIGOS O1sorquIy | 9p 
-=--------- -----~-~-~-~-~---------------- Ng UIs | oF 
---------------------------------- seluoD uene | PP 
w----------------------------- e}JUSUIS[D BuUBNL | Sp 
wr nnn nn ern nnn ene Boy Uegqe sy | 
somata VrSSa wn SE EOE ¥UI) Orpuelely | Ip 
---------~---~---~----~-------- MeBIqeg o1ljoweq | OF 
------------------- BYsl[osuBAG CAnjusAeUENg | 6E 
Sichatatatetedatatatateietaiataiieieiateiatat SJUOIET [LA OUSTepse | BE 
-2---- ------------------------- Z141I9 OJA8qION | LE 
Su sa st SE SSS SSS SSS Sat opleqy Blew | 98 
ao aa naan on === == ---------- Tetaer [epi | Ss 
ponere eeccecerma-ceenceeer BUIqn}A OUIIET[IND | HE 
Renan en ene mn nnn anna n naan n= Blorey OURIDIIOY | $F 
on 2-2-2 --------------------- AorojuOpY unSsNsaY | 7g 
wane anne ------------------ ZNdIO Bl] ep 10o4seg | IE 
Seen towns cananm naan sana eo ae oxselreD osurai0g | 0 
warn enna enn == 2 === -- === ------------ uny 1% | 62 
aa - = 222-2 === -------- 4-5 ------ Byoo1eg uenr | 8Z 
wan rn nn nnn nn nnn nnn naan nanan O8IqO OTEqoy | LZ 


ee eee ee ee ee ee ee 


— aieeomeal 


1914 


rence 


Journal of Sci 


ippine 


a 


The Ph 


26 


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9% “Ady | 9¢ “Ady 
93 “Ady | 9% “Ady 
66 “IVI | 86 “18 
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BBsUBY gpg |---------- 10}1Pq . poses Opssea| aan EY ACTON Alt) Aen {lem c hae go ote ee ear TOIZOID WEIL | 89 
ZaNDSBIA A TZ) |--~--~ uguiaarjog |----- ODieoa saeaee 2) OP dPeattee sass sere c enrages usMedey, using | 19 
Bley BURBS gay |-------~ aer0qeyq |------ LSI) || PS OUI Cal Ly a poen enon onaeuaes “Geman nears uosiiep uenp | 99 
Bun wen 92zp |--------2== OUON FS TSB as maT eT | (GT e i |iemnncin saa an Sea re eZung Buewioig | 99 
s019UOgeL GI6-LI6 |~~~-~~-~- >> auon |----- Opa les Opra-F" PLY >| tna nae ee es ulsne'y ouswele, | 79 
Bllag “loltoyul gpg |~--7 >>> ~~ JeulIe yg |----~ Op -e Opies B2, Wiemiae Slane ae ee uoze’y o1turg | g9 
usdnaeq ‘101193U! Zept |--~--~---- OPteesalFsse= Op apes ss ODESse> Fifa haa cn ee ila ee zenbilug o1oljdurg | 29 
BABII 9F9 | --~ yqusaeatig |----~- OLBWL: |: <S5 LONER [02 | eae acearcnsek oe rate aia uozueg ojqeg | 19 
@pUuES POST |---------—- euoNn |----- Opi asease= i) es OT «| ese aaa yoga oo Saas saa Z@A|B8D PBPIULTT, | 09 
usdnseq “10lt9jUl BIpf |--~---~--~- auoN |---~> opr =aFssae5 ODiadan Co A [ich in aed ia a BZOpUuEeW BIAEqION | 69 
BlINsy Zr |---~- OTIMOSO LT sos oO] SUL | ame ILECT Here |e) fia | tenant ere ouRliBy Buljneg | gg 
Bpey “d pee | ypuLstaatis |------ LB lal [ear OULU Tae | te aa eee eo ee opunurAey asor | 1g 
"BLK 
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‘ponutju0g—anbyjd fo saspo— JI] AIaV 


IX, B, 1 Heiser: Plague in the Philippines oT 


TABLE I1V.—Plague-infected rats. Reported by the Bureau " Science. 
Bocterolpgical examination, positive. 


| No. Where found. Date. How caught. 

1 | 2b1 San Sebastian —__--.-------- Aug. 30,1912 | Poison. 
2 102 Santa Rosas -2 2222. Sept. 6,1912 | Spring trap. 
Sr paloshichapies sete) oe 8 Sept. 6, 1912 Do. 
AA SOO Ma DOnNerOS ==... 8 e Oct. 16,1912 | Poison. 
5 | 157 Estero de Binondo_-__--_--_- Dec. 19,1912 | Spring trap. 
6 | 657 Cabildo (Intramuros) -__-___- Dee. 23,1912 Do. 
7 | 157 Estero de Binondo-_--_---_--_- Dec. 26, 1912 Do. 
8 | 819 Estero Cegado ______-------- Feb. 20,1913 Do. 
On 208 oardinag: 226 oe 2. 22 Bo Mar. 7,1913 Do. 
10 | 857 Eleano_-__-_-_- Mar. 7,1913 Do. 
1) 204: Concha = =-s20-=-...-5 Mar. 9,1913 Do. 
12 | 200 Padre Rada-__-__-_.-_-- Mar. 9,1913 
a3) | 138 Sande- 28 5-5-3 -| Mar. 12,1913 | Club. 
14 | 245 Perla ___ .| Mar. 13,1913 | Poison. 
ApyStOli Corals of. 2.2 oes ae 13, 1913 Do. 
16 | 1316 Sande 16,1913 | Spring trap. 
Ti} 1001 Pesquerra.2- =.= 2 Mar. 19,1913 | Other causes. 
EA 5 2a 0) 6 Col cM aree Bt  e Mar. 26,1913 | Other weapon. 
NOW 420 Sanders a ees = Mar. 29, 1913 | Poison. 
PADS PA Sa) Seb ei aa a eT Mar. 29, 1913 Do. 
PCS Bo UAMIOSP NE: - os — ns Mar. 31,1913 Do. 
22 | 688'T: Alonzo (Int. 1))_....=.--.. Apr. 3,1913 | Spring trap. 
Zeiole Pesqueria- 9-3 0.2825" Apr. 65,1913 | Poison. 
ZA | 1617'Sande (Int. 14) __--_ May 1,1913 | Spring trap. 
PbuiWG49 Sandelnc 2. = 2 Sse a May 6,1913 Do. 
PAT ietetg Wl Ua Cee ae ee ae eed May 8, 1913 Do. 
eUalpOnt bangwaye. po. oe eS May 9,1913 Do. 
28) 207 Velasquez 2222-2222 = 252 Apr. 18,1913 | Club. 
29 | Palumpong__........---..------ May 14,1913 | Unknown. i 
30 | 1811 Juan Luna (Int. 2) ________ May 20,1913 
Si 220; Leben sees 25 a ee May 20,1913 | Spring trap. 
82 Gon Velasquez 2 =) =--<-o--.--_ June 9,1913 Do. 
33 | 447 Conservador (Int.) __-_- ..-- June 15, 1913 | Found dead (with plague). 
S34c, 1908) turan lLunass-=3-2 =. 2<24.2 June 25, 1913 | Spring trap. 
So lea L ONG i Cae eee Se as one | Sept. 6,1913 | Found dead. 
So0roet: Penartibiai 22-0 -52--s22_<-5 Sept. 11,1913 | Spring trap. 
Wis MbSU MVED Ace e eer a ee Sept. 11, 1913 Do. 
88 | Stewart Building_-_..__________ Sept. 19,1913 | Found dead. 

Cat | United States warehouse, Mal- | Nov. 30,1912 | Found sick. 

econ and Pasig River. | 


28 The Philippine Journal of Science 1914 


TABLE V.—Rats destroyed on vessels by sulphur fumigation. 
(Calendar year 1904.) 


Number of rats on— Number of vessels— 


Month. Steam- | Schoon-| River All Having | Having | Fumi- 

ers. ers. craft. | vessels. rats. no rats. | gated. 

January, too a ee 0 0 0 0 0 0 0 
Mebruary 2.2 s22co-eseeeeee 0 0 0 0 0 0 0 
March) 3--2-ccere eee 0 0 0 0 0 0 0 
tAtpril_= oe i ae So 461 25 4 490 38 6 44 
(Mayen. esi. he as 155 106 0 261 51 45 96 
Junes=22 28 ee 153 19 0 172 20 12 32 
duly sao ee oS ee 6 25 0 31 8 20 28 
Aurost - 222-5. = eee 164 9 0 173 7 13 20 
Sentember 5-5. ee 83 0 0 83 1 18 19 
October <-2425:-4--5.4- 255 146 0 0 146 ll 10 21 
INovember a5: 22-2505 5-c5 260 10 0 270 20 15 35 
December: 2... 5-. 225-555 85 9 0 94 8 4 12 
Wot oe eee 1,518 208 | 4| 1,720 164 143 | 307 


TABLE VI.—Kinds of vessels on which rats were destroyed. 
(Calendar year 1903.) 


Vessels. Rats. 
Feats 

Average| 38© ° 

s . Average |. vessels 
A Having | Having i in vessels : 

Kind. Total. Total. in all 6 having 

rats. no rats. wenselal having rath 

rats. 

Steamers 2-525. os encee 168 99 69 1,518 9.00 15. 28 58. 93 
Schoonersecs. scns2 ce eee 134 63 71 203 1.51 3. 22 47.01 
River ‘eraft.23-- 28 6 2 3 4 0.80 2.00 40.00 
All vessels --__-_---- | 307 164 143 1, 720 5. 60 10. 49 53. 42 

| 


TABLE VII.—Rats destroyed on vessels by sulphur fumigation. 
(Calendar year 1903.) 


Number of rats on— Number of vessels— 
Month. Steam- | Schoon-| River All Having | Having | Fumiga- 
ers. ers. craft. vessels. rats. no rats. ted. 
JANUALY: 2. <2. nce soeeees 112 48 0 160 29 23 52 
Bebruary 22. -2..5-- 26 19 0 45 9 26 35 
Mariel 2s .<c4-5eaeceapas 38 16 0 54 9 17 26 
10 14 2 26 7 17 24 
246 10 0 256 19 8 27 
286 5 0 241 15 5 20 
46 85 3 84 12 4 16 
274 18 0 292 13 8 21 
65 24 0 89 14 5 19 
827 9 4 340 9 10 19 
12 al 0 33 13 5 18 
81 6 0 87 14 3 17 
1,473 225 9 1, 707 163 131 294 


IX, B,1 Heiser: Plague in the Philippines 29 


TABLE VIII.—Kinds of vessels on which rats were destroyed. 


(Calendar year 1904.) 
Vessels. Rats. 
Percent- 
A age 
; , Average |.AVerage| vessels 
4 Having | Having * ‘in vessels} having 
Kind. Total. SHE, aheata Total. jin all | Gest Sate 
sels. 
rats. 
———| 
Steamersis = 2225-25-25 -5 154 105 49 1, 473 9.56 13.03 68. 18 
Schooners= 2-62) == -=- 134 54 80 225 1.68 4.17 40.30 
iver craft-2> 2225 -.6--5.2: 6 4 2 9 1.50 2.25 603 

All vessels ___-_----- 294 163 131 1, 707 5.80 10. 47 55. 40 


TABLE IX.—Rats destroyed on vessels by sulphur fumigation. 
’ (Calendar year 1905.) 


- Number of rats on— Number of vessels— 
fy Month. 


Steam- | Schoon-| River All Having | Having | Fumi- 
ers. ers. craft. vessels. rats. no rats.| gated. 
Reva yee oes ke eed 24 15 | 1 40 12 | 6 18 
eebrunry 2-8 4 5. See 53 re gh; ) 56 8 / : 17 
UT SiR ee 141 23 0 164 19 13 32 
V0 nl eae Se ee 125 18 0 143 17 4 21 
Migs 8 oe cee ee 22 3 0 25 5 8 ie 
UGG = Se eee eee 96 26 0 122 | 12 | 3 15 
| ul 0 3/ un 3| 19 
oe | 23 0 0 23 | 4 | 4 8 
September. ==... -2<-2:.-' | 69 0 0 69 5 | 2 ui 
Ginter ersten ons 5 ee | 35 | 9 0 44 13 2 15 
oven er eee ness aaa 18 3 | 0 21 | 6 1 7 
ecember_ a... a--5----2--- i) 0| 0 0 | i) 0 | ) 
60 172 


TABLE X.—Kinds of vessels on which rats were destroyed. 
(Calendar year 1905.) 


Vessels. Rats. | 


Per cent- 
| = age ae 
| x : Average |. verage| vessels 

. | Having | Having es lin vessels) having 
Kind. Total. rats. | no rats. | Total. naan having rats. 
/ | ¢ rats. 
| | | 

teaImerdiosn a9 otc ee ake, 109 72 37 | 678 6.22 9. 42 66.05 
Schoonerq==- 255.55.) 62 39 23 111 1.80 2.87 62. 90 
River.crafp <-05 23.2282 1 1 0 | 1| 1.00 1.00 | 100.00 
All vessels ___....--- | 172 112 | 60 | 790 | 4.60 7.05 65.11 


30 


The Philippine Journal of Science 1914 
TABLE XI.—Rats destroyed on vessels by sulphur fumigation. 
(Calendar year 1906.) 
Number of rats on— Number of vessels— 
Month. Steam- | Schoon- | River All Having | Having | Fumi- 
ers. ers. craft. | vessels. rats. no rats. | gated. 
January 22 ee 10 0 0 10 1 0 af 
Mebruanyae- eee 0 0 0 0 0 1 1 
10 5 0 15 3 0 3 
0 0 15 15 1 2 3 
2 0 0 2 1 a! 2 
169 2 0 171 19 0 19 
205 0 0 205 14 0 14 
56 9 0 65 12 1 13 
7 23 0 30 3 0 3 
October see ee 58 12 0 70 8 1 9 
November! s2ccs<-2 25 23 0 0 23 2 0 2 | 
December. 25S aes 8 | 1 0 9 3 0 3 
Mota) sues sees 2 ves 548 52 15 615 67 | 6 73 
i 
TABLE XII.—Kinds of vessels on which rats were destroyed. 
(Calendar year 1906.) 
| Vessels. 4 Rats. Bartlty - 
| Average| 2®&¢ 4 
F | Having | Having Average |i, vessels| Vessels 
Kind. | Total. Sip rigwatn Total. Sa a having having, 
Tee | 
}) Steamers 2 -- 2s an 59 55 4 548 9.30 | 9. 98 93. 22 
|| Schoonerst:---. tc 13 ll 2 52 4.00 4.73 | 84.61 
Rivercraft 25.2 eee | 1 1 | 0 15 15. 00 15.00 | 100.00 
All vessels. --_-____- | 73 | 67 | 6 615 8. 42 | 9.18 91.78 
i 


TABLE XIII.—Rats destroyed on vessels by sulphur fumigation. 
(Calendar year 1907.) 


Number of rats on— 


Month. Steam- | Schoon- 
ers. ers. 

SVAMUAY Y= - He san or 20 1 
Mebriary — = 2-2 ee ae 67 12 
Marchi ost ae. p eee 110 11 
(AS YIE - 22 Res ees 262 14 
May so sacar 66 6 
WINES = 25 ee 28 0 
DW asi Sn eee 38 0 
Aneuat| A. o207 non eos 24 | 0 
September ......=-+-=---s2 0 13 
October. 252-2... es 37 2 
November 22o-2222. 22-2 51 10 
December: ==... 25.5225.—4 14 4 
| Totals 3". soe 22) 717 73 


River 
craft. 


All 
vessels. 


Number of vessels— 


Having | Having | Fumi- 
rats. no rats. | gated. | 

2 0 2 

8 1 9 

26 al 27 

18 0 18 

10 1 ll 

2 1 3 

5 8 13 

4 2 6 

1 3 4 

7 5 12 

12 2 14 

4 0 4 

99 24 123 


Heiser: Plague in the Philippines 81 


TaBLe XIV.—Kinds of vessels on which rats were destroyed. 
(Calendar year 1907.) 


Vessels. Rats. 
Percent- 
| age of | 
Kind. Total. Having | Having Total. seuenane in vessels} having 


rats. no rats. having rats. 


Average| vessels | 

vessels. Patel | 

SLCHINCYS Sasa essen ae 91 77 14 717 7.88 9.31 84.61 | 
Schoonerss— <-> 05 --ncn5 25 20 5 13 2.92 3.65 80. 00 
River craft.--:--=-.-.----- T 2 5 41 5.85 20.50 28.57 


TABLE XV.—Rats destroyed on vessels by sulphur fumigation. 
(First half of calendar year 1908.) 


Number of rats on— Number of vessels— 

Month. | 
Steam- | Schoon-| River All Having | Having | Fumi- 
ers. ers. craft. | vessels. rats. no rats. | gated. 
re ee 188 3 0 191 9 0 9 
INE CC Se ey | 118 7 0 | 125 10 1 ll 
2) Sei teen ere 59 2 0 | 61 5 0 5 
ee ERI IOA, Fe 5 oa, 16 0 2 18 2 2 4 
‘eee eee 17 2 0 19 4 1 5 
Pe A te a! 91 6 0 97 8 1 9 
| 20 2 | 511 | 38 5 43 


TABLE XVI.—Kinds of vessels on which rats were destroyed. 
(Calendar year 1908.) 


Vessels. Rats. 
Percent- 
mn age of 
verage| vessels 
Kind Total Having | Having Total eTEES in vessels} having 


having rats. 


rats. no rats. 
vessels. mre 


MteAINenAssce ses pees 31 27 4 489 15.78 18.01 87.10 
Nehooners= = --22— 5-552. -- ll 10} « a 20 1.82 2.00 90. 90 
iver crafts 20 6 8, 1 1 0 2 2.00 2.00 | 100.00 

All vessels _________- 43 38 5 511 | 11.88 | "48,45 | 88.37 


32 


The Philippine Journal of Science 


1914 


TABLE XVII.—Rats destroyed on vessels by sulphur fumigation. 


Month. 


(Fiscal year 1909.) 


Number of rats on— 


Number of vessels— 


Steam- | Schoon-| River All. Having | Having | Fumi- 
ers. ers. craft. | vessels. rats. no rats. | gated. 
152 74 0 DOG NE Bus she | Vee ne 38 

73 0 7 BO) Re ore. 2 ee ae 14 

24 0 0 De ee aN hi ek eS 9 

29 4 0 SB eS eee [ead rece 8 

30 0 0 80); soeee eee 5 

42 0 0 Ou Smee ere 3/2 eee eee 8 

7 4 0 P| he ee well 5 

22 0 0 POM ag i Ry Pas ES 7 

51 0 0 153 Ul (ea ee eee ee | 8 

38 2 0 2 AUN Rae aS Se Hse es oy 9 

45 6 0 isi [ER aes See ee ee FO ERS 9 

116 0 0 VIG ecesne ashe d 6 
629 90 | 7 (coe eee 126 


TABLE XVIII.—Kinds of vessels on which rats were destroyed. 


Kind. 


(Fiscal year 1909.) 


Vessels. 


Total. 


Having | Having 
rats. no rats. 
76 32 

12 5 

1 0 

89 37 


Rats. 
Percent 
Average | 28¢ © 
rota, | ANeTRE*|in'vessete) yoesels 
"| vessels having ro 
fs rats. rats. 
629 5. 82 8.30 70. 53 
90 5.29 7.50 70. 60 
if 7.00 7.00 | 100.00 
726 5.76 8.15 70.63 


TABLE XIX.—Rats destroyed on vessels by sulphur fumigation. 


(Fiscal year 1910.) 


Month. 


Number of rats on— 


Steam- | Schoon-| River 
ers. ers. craft. 

0 0 

0 0 

7 0 

0 0 

0 0 

0 0 

16 0 

0 9 

9 31 

67 5 

0 0 

0 5 

99 50 


Number of vessels— 


All ves- | Having | Having | Fumi- 
sels. rats. no rats. | gated. 


Heiser: Plague in the Philippines 33 


TABLE XX.—Kinds of vessels on which rats were destroyed. 
(Fiseal year 1910.) 


Vessels. Rats. 


Percent- 

| mn age ee 

; verage vessels 

3 Having | Having Average in vessels having 
Kind. Total. rats. no rats. Total. pola | having rats. 

rats 

pee eae ee oe | 109 88 21 1, 634 15.00 18.57 80.7 
Voialenlee See he ere 32 16 16 | 99 3.10 6.20 50.0 
18 BO «| ee @i| 18 24 50; 1.20; 2.977|  43.0 


JY ae eee | 183 | 122 61 1,783 | 9.74 | 14.62} 66 


TABLE XXI.—Rats and mice destroyed on vessels by sulphur fumigation. 
F (Fiscal year 1911.) 


| Found in— 
Steamers. ' Schooners. Launches. | All vessels. | 
Month. | 
Rats Rats Rats | | Rats 
Mice. Rats.| and Mice.| Rats.| and | Mice.| Rats.) and | Mice. Rats. | and 
mice. mice. | mice. mice. 
109| 158 
202 274 
230 | 282 
134 147 
45 | 68 
85 | 50 
647 | 724 
136, 204 
78 | 161 
277 j 303 
115 237 
172 202 
2,080 | 2,795 


TABLE XXII.—Kinds of vessels on which rats and mice were destroyed. 


(Fiseal year 1911.) 
| Average number i | 


Number of Average number 


vessels. Number of— in all vessels. ip. es Percent- 
2 ge 
ene (Grn) ea —— = ; vessels 
Hay-| Hav- Rats | Rats | Rats hayane 
Total.| ing |ingno Mice.| Rats.| and | Mice.| Rats.| and | Mice.| Rats.) and " 
rats. | rats. mice. mice. mice. 
Steamers a 170 | 145 25 | 526 |1,802 |2,328 | 3.09 |10.60 13.69 | 3.63 |12.43 |16.06 | 85.300 
Schooners---_ 641 62 ZY 165 | 181 | 336 | 2.42 | 2.83 | 5.25 | 2.50 | 2.92 | 5.42 | 96.875 
Launches -_--| 33 32 | I 34 | 97 | 181] 1.03 | 2.94 | 3.97 | 1.06 3.03 | 4.09 | 97.000 
SS SS 
All ves- | 
sels ___| 267 | 239 28) 715 fa 080 |2, 795 | 2.67 | 7.79 10.46 | 3.00 | 8.70 |11.70 | 89.500 


« 124293——3 


34 


The Philippine Journal of Science 


1914 


TABLE XXIII.—Rats destroyed on vessels by sulphur fumigation. 


(Fiscal year 1912.) 


Vessels fumi- 


Found in— pated 
Steamers. Schooners. River craft. All vessels. ‘ 
Month. ou : 2 | 
cs) | lg cs] oe) H : 

i) ae Cae | a2 3 | si eg 3 a a § 2 E 3 

S| Bese es Peele eee) 2 1 8 1 S8| e | ome 

=f aU ==2 -=  =g=  V 

uly: = 22 51] 165 | 216 6| 49) 55 4] 16} 20 61 |) 230; 291} 24 5 29 
August ___| 125 211 | 336 5 9} 14] 14] 11] 25) 144] 231] 3751] 40 6 46 
September} 51 98; 149) 37) 46] 83 4 fhyee ak 92)| 15 | 2431) 25 9 34 
October ___| 66 7 | 141] 38 0 3 0 0 0 69 75 | 144] 11 2 13 
November | 46 87 133 | 174 | 32 | 206 0 1 1} 220; 120} 340; 16 ik ily/ 
December_| 45 174 | 219 1 7 8 0 0 1 46 | 181 | 227 13 qT 20 
January ==} 22 108 130 | 17 16} 53| 6] 14} 20 45 | 158! 203] 36] 11 47 
February _| 100 193 293 9 10; 19 | 0 0 0 109 | 203] 312] 22 5 27 
March __-__| ° 84 | 459) 543] 3 } 5] 8 | 4 5 9 91} 469} 560] 29) 10 39 
Aprilus-3. 70 } 256 | 326 10; 16 | 26 3 0 3 83 | 272} 355] 18 6 24 
Mays ss = 3 27 | 170 197 | 40} 38 | Titans al 4 5 68} 212] 280! 25 4 29 
June .....- | 25] 87) 62] 0] 8] 8] 4/7] m| 29! 52] s1| a3| 7| 20 
Total__| 712 2, 033 2» 745 | 305 | 256 | 561 | 40} 65 | 105 |1,057 2,354 poems 272) %8 | 345 

L a ~ 3 1 


TABLE XXIV.—Kinds of vessels on which rats and mice were destroyed. 


(Fiscal year 1912.) 


Kind of vessel. 


Steamers___-_ 
Schooners___ 


River craft _ 
a All vessels_ 


Average number 
in vessels having 


rats. Percent- 


Number of te Average number 
vessels. Number of in all vessels. 

Hay- Hay: Rats Rats 
Total.| ing et Mice.| Rats.| and | Mice.) Rats.| and | Mice. 
rats. 3 mice. mice. 

rats. 
223 | 175 48 | 712 |2,033 |2,745 | 3.19 | 9.12 |12.31 | 4.07 
72 57 15 | 305} 256} 561 | 4.24 | 3.55 | 7.79 | 5.35 
50 40 10 40 65 | 105 | 0.80 | 1.30 | 2.10 | 1.00 
345 | 272 73 |1, 057 rice (3, 411 3.06 | 6.82 | 9.88 | 3.89 


age of 
vessels 
having 


Rats.) and | *2ts- 


mice. 


11. 62 |15. 69 
4.49 | 9.84 
1.62 | 2.62 


78. 48 
79.17 
80. 00 


8.65 |12.54 | 78.84 


_ uM ee ee ee ee ee ee 
ee 


-— 
~ 


x Bt: Heiser: Plague in the Philippines 385 


TABLE XXV.—Rats destroyed on vessels by sulphur fumigation. 
(Fiscal year 1913.) 


Found in— 
: Steamers. Schooners. . River craft. All vessels. 
Month. ee EST SE 
Mice.) Rats. ae Mice.| Rats. ad Mice.| Rats. aa Mice.| Rats. Hetsand 

mice. mice. mice. ‘ 
Bhuyee se seen 71 114 185 11 23 34 0 6 6 82 143 225 
August_-_.------ 85 84 119 0 0 0 0 2 2 35 87 2121 
September ----- 39 154 193 16 21 87 0 12 12 55 189 b242 
October -------- 18 145 223 4 16 20 37 89] 126] 119 250 369 
November-.--_--- 149 82 231 12 12 24 30 | 103; 1838] 191 197 388 
December ------ 21 270 291 0 il 1 12 67 79 33 338 371 
January -----.-- 1 12 13 A 0 3 1 6 7 5 18 23 
February ------- 6 44 50 0 1 1 1 0 1 7 45 52 
‘onsen 19 25 44 3 1 4 0 0 0| 22| 26 48 
Se eee 14 150 164 0 5 5 0 20 20 14 175 189 
oe eee 26 178 204 0 12 12 0 3 3 26 198 219 
Bee wectiea 3 21 140 161 2 3 5 Hf 7 30 143 178 
eee 480 | 1,398 | 1,878 51 95 | 146 88 | 308] 396 | 619 | 1,804 | 2, 428 


® Plus 1 found on wharf. > Plus 2 found on wharf. 


TABLE XXVI.—Kinds of vessels on which rats and mice were destroyed. 
(Fiscal year 1918.) 


Number of a | Average number | AV¢r#ge number 
vessels. Number of in all vessels. | ™ vaeHels Baving Percent- 
E age of 
Kind of vessel. vessels 
Hav-| Hav- Rats Rats | Rats | having 
'Total.| ing |ingno| Mice.| Rats.) and | Mice.| Rats.) and |,Mice.| Rats.! and | rats 
rats. | rats. mice. mice. mice. 
Steamers_...| 225} 178 62 | 480 |1,398 |1,878 | 2.13 | 6.21 | 8.34 | 2.75 | 8.08 |10.83 | 76.89 
Schooners..-| 53 29 24 51 | 95] 146 | 0.96} 1.79 | 2:75 | 1.76 | 3.27 | 5.038 | 54.72 
River craft -| 320} 129] 191 88 | 308 | 396 | 0.27 | 0.96 | 1.23 | 0.68 | 2.39 | 3.07°| 40.30 
_ Allvessels-| 598 | 3831] 267] 619 |1,804 |2,423 | 1.08 | 3.02 | 4.05 | 1.93 | 5. 62 7.55 | 53. 68 
i 


TABLE XXVII.—Rats destroyed on vessels by sulphur fumigation. 
(January 1, 1903, to June 30, 1913.) 


Number of vessels. Rate. 
Hereent: 
“| age o: 
BenAIOF veseel: | Having | Having pes ‘number reseks 
V1 = i 
Total. | “rats. | norats. | Total- |in all ves- pee eee eens 
sels. mare 

Steamersiac=. so sae. ee 1, 447 1, 092 355 14, 632 10. 11 13. 40 15.47 
DSehooners=2* 2-2-2 so-nce 617 373 244 1,916 3.10 5.14 60. 45 
River craft: 2.2: 2-2-5. 467 231 236 761 1.63 3.30 49.46 


| All vessels _..-..-.-- 2,531 1, 696 835 | 217,312 6. 84 10. 80 67. 00 


* Three found on wharf. 


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fiom fh ¥ Fae 
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7a. 


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te aye veskcnel 


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ILLUSTRATION 


Lap of Manila, showing the location of plague cases in 1912-1913. 


7 


HEISER: REAPPEARANCE OF PLAGUE.] 


ting 


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BA 


7 yy { 
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MANILA, SHOWING 


MAP lI. 


pete. Pui. Journ. Scr., IX, B, No. 1. 


Sampatoc 
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a. TA — SHOWING PLAGUE 
: 1912~1913 

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— Sanitary Map 
_ Bureau of HEALTH 


7? ae < 1% i. iy 


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A PRELIMINARY REPORT OF EXPERIMENTS ON THE CULTIVA- 
TION OF THE VIRUS OF RINDERPEST IN VITRO 


By WILLIAM HUTCHINS BOYNTON 


(From the Veterinary Division, Bureau of Agriculture, Manila, P. I.) 


The literature on rinderpest contains numerous reports of 
investigations carried on with the object of isolating the caus- 
ative agent of that disease. Tartacovsky(1) gives a thorough 
review of the results obtained by numerous workers up to 1896. 
Nencki, Sieber, and Wijnikewitch(2) in 1898 published a paper 
on the etiological agent of rinderpest, but their conclusions have 
not been verified. They did, however, report success in main- 
taining the virus in vitro to the fourth generation. No explicit 
statement was made regarding the time covered by these four 
generations, but the writers observed that it is best to transfer 
cultures every three days. Koch(3) in 1897 in the second report 
of his investigations in South Africa on the etiology of rinderpest 
states that all his efforts to isolate and cultivate the virus of 
rinderpest were fruitless. 

In the present report there will be given merely a summary 
of the preliminary experiments which have been carried on at 
the veterinary research laboratory at Alabang and which have 
led up to the present work. 

It was observed from various experiments that the causative 
agent of rinderpest remained alive and maintained its virulence 
much longer under anaérobic than when under aérobic con- 
ditions. The medium used in these earlier experiments was 
principally virulent blood drawn under aseptic conditions from 
animals suffering with the disease. It was noted that blood 
drawn from an animal in the early stages of the disease—that 
is, from one to three days after the initial rise of temperature— 
maintained its virulence much longer than blood drawn from 
an animal in the later stages of the disease, near the point of 
recovery or of death. Undoubtedly the antibodies present in 
the samples of blood play an important réle in the last-mentioned 
condition. 


* Archibald R. Ward, chief. 
39 


40 The Philippine Journal of Science 1914 


The next step was to locate, if possible, the blood element 
which harbored the virus. For assistance in this work, I am 
greatly indebted to Dr. M. A. Barber of the Bureau of Science. 
Doctor Barber used his pipette method for the isolation of single 
microérganisms to pick out and separate the different blood 
elements. 

Two susceptible animals received, respectively, 200 and 255 
red blood cells from an animal sick with rinderpest. The 
animals were unaffected by the injections, and at a later date 
they were proved to be susceptible. Two animals were injected 
with 15 and 40 leucocytes, respectively, obtained from the blood 
of infected animals. They were unaffected by the injections, 
but later were proved to be susceptible. Three animals were 
injected with blood platelets. They received, respectively, 
6,000, 770, and a large number, the exact count not determined. 
These animals remained well, but later were proved to be sus- 
ceptible. The blood from which these elements were taken was 
checked in each case and found to be virulent. 

The question then arose, does the virus occur as thickly dis- 
tributed in the blood of an infected animal as was previously 
supposed. Therefore, it became necessary to determine the 
smallest dose of whole blood that would cause the disease. Fine 
capillary pipettes were made, and a certain length was marked 
on the glass. The weight of mercury filling this designated 
portion was compared with the weight of 1 cubic centimeter 
of mercury. In this way the capacity of the minute quantity 
defined by the mark on the capillary tube was approximately 
obtained. It was found that %o7) cubic centimeter of virulent 
blood transmitted the disease to a susceptible animal, but %ogo 
cubic centimeter and 0.0001 cubic centimeter of virulent blood 
failed to transmit the disease. The susceptibility of these ani- 
mals was proved later. These results gave an approximate 
indication of the minimum amount of virulent blood which would 
transmit the disease and were an aid in the culture work for 
comparison with the dilution of the original blood brought about 
by successive transfers in culture media. 

The final step was to find a medium in which the virus would 
remain alive and multiply. Many unsuccessful attempts were 
made using various kinds of media. The virus would either die 
or at least lose its virulence in from thirty-six to sixty-eight 
hours. ~In no case was the second tube of medium shown to 
contain virus. Finally, apparently positive results were obtained 
by modifying slightly the medium described by Nencki, Sieber, 


= a” 
e 


IX, B, 1 Boynton: Cultivation of Virus of Rinderpest Al 


and Wijnikewitch(2) and with the medium used by Bass and 
Johns.(4) This report is based upon work with these modified 
media. 

TECHNIQUE 


The first medium tested was a salt-peptone preparation advo- 
cated by Nencki, Sieber, and Wijnikewitch. It is composed of 
900 cubic centimeters of water to which are added 100 grams of 
peptone Witte and 20 grams of sodium chloride. The mixture 
is filtered, placed in test tubes, and sterilized. I have been 
unable to obtain successful results in the cultivation of rinderpest 
virus in the medium described. This medium was modified by 
adding 0.1 cubic centimeter of a 334 per cent solution of glucose 
to each 10 cubic centimeters of the salt-peptone mixture de- 
scribed by Nencki, Sieber, and Wijnikewitch. Test tubes 1.5 
centimeters in diameter and 15 centimeters long were employed. 
The glucose solution to the amount of 0.1 centimeter was added 
to each tube, and to this were added 10 cubic centimeters of 
the salt-peptone solution. It was sterilized for one hour in an 
autoclave at 185° C. on the day previous and one-half hour just 
prior to inoculation. Normal blood was drawn under aseptic 
conditions from a nonimmune animal and defibrinated by shaking 
with glass beads. One cubic centimeter of this blood was added 
to each tube immediately before it was inoculated. Blood from 
an animal suffering with rinderpest was then drawn under 
aseptic conditions and defibrinated. Each tube was inoculated 
with either 0.5 or 1 cubic centimeter of the infective blood. 
Anaérobic conditions were produced by covering the surface of 
the inoculated media with from 1.5 to 2 cubic centimeters of 
sterile paraffin oil. The tubes were then placed in the incubator 
at 40° C. 

In making transfers, a sterile pipette of 1 cubic centimeter 
capacity was inserted into the culture tube from which the 
transfer was to be made and the medium was thoroughly agitated 
by filling the pipette with the medium and emptying. This was 
repeated four or five times. Then either 0.3 or 0.5 cubic centi- 
meter of the culture was transferred to the prepared culture 
medium, after which the tubes were sealed with paraffin oil 
and placed in the incubator. It was found best to make these 
transfers every three or four days. 

The medium which has given the best results is a modifica- 
tion of the one used by Bass and Johns for cultivating the 
plasmodium of malaria. To each of a series of test tubes was 


42 The Philippine Journal of Science 1914 


added 0.1 cubic centimeter of a 334 per cent glucose solution. 
These tubes were then placed in the autoclave and sterilized. 
Ten cubic centimeters of normal defibrinated blood from a 
susceptible animal were then added to each tube, which was 
immediately inoculated with 0.5 cubic centimeter of defibrinated 
blood from an animal suffering with rinderpest. Each culture 
was then covered with from 1.5 to 2 cubic centimeters of sterile 
paraffin oil, and was placed in the incubator at 40° C. 

In making transfers, a sterile pipette of 1 cubic centimeter 
capacity was inserted into the medium from which the transfers 
were to be made and the culture was agitated by filling the 
pipette with medium and emptying several times. Either 0.3 
or 0.5 cubic centimeter of the culture was transferred into the 
blood-glucose tubes, which were then sealed with sterile paraffin 
oil and placed in the incubator. 

I have found it best to make transfers every three or four 
days as the virus in culture media has a tendency to lose its 
virulence or die after remaining several days in one tube. 

In one series in salt-peptone mixture the virus in the primary 
tube of culture medium lost its virulence on the sixth day, while 
I was able to carry the same strain alive through four transfers, 
covering a period of thirteen days. 

During the months of July, August, and September, 1913, 
I have twice succeeded in carrying the virus in virulent form 
in the glucose-blood medium to the sixth transfer, covering 
periods of nineteen and twenty-one days, respectively. I have 
twice succeeded in carrying the virus in virulent form in the 
salt-peptone mixture to the fourth transfer covering periods of 
twelve and thirteen days, respectively. 

In one series the fifth transfer from glucose blood medium 
was carried into the salt-peptone mixture which was allowed 
to incubate three days and proved virulent when injected into 
a susceptible animal. These six transfers in culture media 
cover a period of eighteen days after the virus was taken from 
the infected animal. 

The question then arose whether there was really a multiplica- 
tion of the virus in these media or whether the original blood 
was transferred from one tube to another up to the sixth transfer 
in sufficient quantity to cause the disease when injected into a 
susceptible animal. In one of the series in which the sixth 
transfer proved virulent, 0.3 cubic centimeter of the culture was 
used in making the transfer each time. In computing the dilu- 
tion of the original virulent blood in the sixth transfer, it was 


x%,B,1 Boynton: Cultivation of Virus of Rinderpest 43 


found to be approximately %4.3090, cubic centimeter. From the 
experiments quoted above it was found that %o.) and 0.0001 
cubic centimeter of fresh virulent blood failed to transmit the 
disease. Therefore, it appears that there was a multiplication 
of the virus in the culture tubes, since the dilution of the original 
virulent blood that resulted from the successive transfers was 
so much greater than the quantity which was found necessary 
to produce the disease. In one series I was not able to reproduce 
‘the disease from the medium first inoculated after twelve days, 
but the fourth transfer corresponding to a period of twelve days 
after removal from the sick animal proved virulent. This would 
suggest that the virus continued to live in the transfers, but 
died out, or lost its virulence, in the original tube of culture 
medium. 

In the salt-peptone medium myriads of minute bodies could 
be seen under dark field illumination, especially in transfers of 
the first and second generations. A large number of check 
examinations have been made with normal blood. In some 
instances bodies practically identical with those found in cultures 
made from virulent blood are found. Therefore, at present, 
nothing can be stated concerning the morphology of the etio- 
logical factor. In the salt-peptone medium there appears a per- 
ceptible cloudiness above the blood on the bottom of the tube. 

Up to the present time the best results have been obtained by 
taking blood from an animal in the early stages of the disease; 
that is, about the second day of temperature. All animals em- 

’ ployed to test the existence of rinderpest virus in culture media 
were kept under such conditions as to warrant the belief that 
no accidental infection occurred to cause fallacious conclusions. 

The greatest precaution must be taken against contamination, 
since the media are composed chiefly of raw blood which cannot 
be sterilized. I have twice lost the seventh transfer on account 
of bacterial contamination, which appears to kill the virus in 
a very short time. The presence of contaminative bacteria is 
usually revealed by a pellicile immediately beneath the paraffin 
oil. 

One animal which was inoculated with the fourth transfer 
recovered from the disease. This animal is at present being 
hyperimmunized with cultures, with the intention, in the near 
future, of testing the potency of its serum as compared with 
serum from animals immunized with virulent blood. Experi- 
ments are being carried on to determine if it is possible to 
produce a toxin in culture and to prepare an antitoxin. Work 


44 The Philippine Journal of Science ar 7 


is also under way to determine the practicability of substituting 
cultures of rinderpest virus for virulent blood in simultaneous 
inoculations. 

The results obtained present a wide range of possibilities for 
the improvement of the present technique of immunization and 
for the determination of the etiological factor, which may revise 
the present methods of combating the disease. 


SUMMARY 


1. From the results obtained from various experiments it is 
evident that the virus of rinderpest requires either partial or 
complete anaérobic conditions for its existence. 

2. The virus of rinderpest has been carried in virulent form 
in two separate series up to the sixth transfer in glucose-blood 
culture media, covering periods of nineteen and twenty-one days, 
respectively. 

3. In one series the medium first inoculated was nonvirulent 
at the end of twelve days, while the fourth transfer from this 
tube of culture medium after the same period of time was 
virulent. 

4. Results obtained from numerous experiments indicate that 
fresh blood from nonimmune cattle as a main constituent and 
glucose as an addition are essential components of the culture 
media. 

REFERENCES 


(1) Tartacovsky, M. M. Contribution 4 l’étiologie de la peste bovine. 
_ Arch. sci. biol., St. Petersbourg (1896), 4, 295-327. 

(2) NeNckKI, M., SreBer, N., and WIJNIKEWITCH, W. Recherches sur la 
peste bovine. Jbid. (1896), 4, 374-896. 

(3) Kocu, R. Reise-Berichte tiber Rinderpest, etc. Julius Springer, Berlin 
(1898), 5-33, 56, 65; Report. Veterinary Journ. (1897), 45, 204; 
Berichte des Herrn Prof. Dr. Koch tiber seine in Kimberley gemachten 
Versuche bezitiglich Bekamfung der Rinderpest. Centralbl. f. Bakt. 
etc., 1 Abt. (1897), 21, 526. 

(4) Bass, C. C., and JoHNS, F. M. The cultivation of (Plasmodium vivax 
and Plasmodium falciparum) in vitro. Journ. Exp. Med. (1912), 
16, 567. 


AN ATYPICAL CASE OF RINDERPEST ‘IN A CARABAO? 


By WILLIAM HUTCHINS BoyNTON 
(From the Veterinary Division,’ Bureau of Agriculture, Manila, P. I.) 


One chart 


In studying the subject of rinderpest in the textbooks one is 
apt to gain the idea that this affection is infallibly an acute, 
febrile, infective disease. Descriptions usually state that the 
first symptom noticeable is a rise of temperature, which appears 
in from three to five days after infection. Some observers have 
noticed a rise in temperature as early as from thirty-six to 
forty-eight hours after infection. The symptoms which follow; 
such as, a seropurulent discharge from the nostrils and eyes, 
diarrhea, loss of appetite, emaciation, and general debility, 
usually appear in from one to four days after the rise of 
temperature. 

The more recent periodical literature contains references to 
the fact that rinderpest occasionally assumes a mild type, be- 
coming very difficult to recognize. 

Littlewood(1) in Egypt has observed that cattle imported from 
Asia Minor may not show clinical symptoms and yet at autopsy 
reveal lesions of rinderpest. 

Rickmann(2), writing of rinderpest in German Southwest 
Africa, refers to the fact that cattle and other animals may be 
infected to an imperceptible degree. 

Eggebrecht(3) observed in China, that some animals infected 
with rinderpest show*no visible signs of the disease beyond a 
rise in temperature to 40° C. or higher for two days. 

Baldrey (4), describing conditions in India, states that by long 
residence of any organism of contagious animal disease in one 
place the disease becomes weakened in virulence to the animals 
of that place. Thus, animals infected with rinderpest may act 
as carriers without showing symptoms. 

In the course of work on rinderpest, I encountered a case which 
yielded some interesting facts regarding mild symptoms and 
transmissions by both blood injection and by close association. 

Carabao 3235, upon which the observations made in this paper 
are largely based, was received at the veterinary research labo- 
ratory at Alabang, Laguna Province, Luzon, for experimental 


*To be published as Bulletin No. 31 of the Bureau of Agriculture of the 
Government of the Philippine Islands. 
? Archibald R. Ward, chief. 
45 


A6 The Philippine Journal of Science 1914 


purposes on August 35, 1911, having come from Davao District, 
Mindanao. It was kept in quarantine with the other experi- 
mental animals until inoculated on September 23, 1911. At that 
time this animal and bull 3205 were each given 10 cubic centi- 
meters of virulent blood from an animal sick with rinderpest. 

Nine days after inoculation, 3235 was noticed to have a diar- 
rhoea and the eyes were slightly congested; at ten days the 
diarrhoea became profuse, seropurulent exudate was discharged 
from the eyes and nostrils, and the animal ate but little. After 
eleven days the animal stopped eating and displayed all the 
external appearances of a severe case of rinderpest, except that 
the temperature had not at any time gone higher than 38°.9 C. 
On the twelfth day the temperature dropped to 36°.4 C., which 
is subnormal, and the animal died during the night of the twelfth 
day after inoculation. Post-mortem examination revealed large 
ulcers in the mouth and marked congestion of the peritoneal 
cavity, fourth stomach, and rectum. The duodenum was also 
markedly congested, and showed many small ulcers. The cecum 
was but slightly congested excepting around the ileocxecal valve. 
Thus the lesions were regarded as typical of those of rinderpest. 

From the temperature chart of 3235 it can be noted that the 
temperature never rose above normal and was a little lower than 
that of the average healthy animal. On the fifth and sixth day 
there was a variation in the temperature which is often seen 
in rinderpest just before the initial rise. However, the only 
typical evidence of rinderpest so far as temperature is concerned 
was the drop to subnormal on the eleventh day, at the time of 
collapse. , 

There is no doubt that animal 3235 was suffering from rin- 
derpest, for 3205, which was inoculated with the same blood, 
experienced a typical attack. The identity of the disease in 
3235 is further shown by a series of inoculations of its blood, 
at various stages of the attack, into other animals. Susceptible 
animals were inoculated with blood at three, four, five, six, and 
eleven days after the original inoculation of 3235. Of these, 
5 animals contracted typical attacks of rinderpest and 3 died, 
and post-mortem examination showed typical lesions of rinder- 
pest. No injections were made from the seventh to tenth days 
after injection because at that time the absence of the febrile 
temperature in 3235 led to the conclusion that the animal did 
not have rinderpest. Susceptible animals were exposed in the 
same pen with 3235 on the eleventh and twelfth days without 
contracting rinderpest. Another susceptible animal was exposed 
for twenty-four hours in the same pen in which 3235 had died 


——— eS 


“1x,B,1 Boynton: Rinderpest in a Carabao AT 


the day before, but did not contract rinderpest. The suscepti- 
bility of these three animals to rinderpest was proved later by 
their becoming infected with the disease on exposure in other 
experiments. 

CONCLUSIONS 


1. From the facts of this case, the evidence is conclusive that 
an animal may experience a fatal attack of rinderpest without 
the occurrence of a rise in temperature. 

2. The blood of carabao 3235 was shown to be infected within 
forty-eight hours after it was originally injected with virulent 
blood. 

3. It was shown that the blood was virulent on the eleventh 
day when injected into a susceptible animal, yet exposure to the 


No. 3235. 


Eg 


let req | | [| | 
lel | 


[>I 
SI 
|_| 
ol 
| 
ic] 
fa 


same animal from which blood was drawn did not cause rinder- 
pest in the exposed susceptible animal. 

4. With regard to the three animals which failed to contract 
rinderpest by exposure, the question is raised as to whether 
rinderpest spreads by contact readily in the later stages of the 
disease or whether the disease must necessarily be accompanied 
by a rise of temperature before it can be spread by contact. 


REFERENCES 


(1) LiTTLEWoop, W. Cattle plague in Egypt in 1903-04-05. Journ. Comp. 
Path. & Therap. (1905), 18, 312. 

(2) RICKMANN, W. Tierzucht und Tierkrankheiten in Deutsch Suedwest 
Africa (1908), 156. 

(3) EGcrBRECHT, M. Untersuchung iiber die Rinderpest in Ostasien. 
Zeitschr. f. Infektionskrankh. etc. (1910), 7, 54. r 

(4) Bauprey. Climatic influence upon the incidence of disease. Agr. 
Journ. India (1912), 7, pt. III, 294. 


EXPERIMENTS UPON THE TRANSMISSION OF RINDERPEST ! 


By ARCHIBALD R. WARD, FREDERICK WILLAN Woop, and WILLIAM 
HUTCHINS BoyNTON 


(From the Veterinary Division, Bureau of Agriculture, Manila, P. I.) 


Two plates and six charts 


In combating rinderpest, information concerning the length 
of time the virus remains active outside of the body under various 
natural conditions is of great importance in suggesting the 
measures to be employed in the field. Information concerning 
the period during the course of the disease when the virus is 
disseminated by sick animals is of equal usefulness. 

The literature on the disease consulted by us contains scanty 
and contradictory reference to these significant topics. 

Réfik-Bey and Réfik-Bey (1) state the following: 


Infected areas do not remain dangerous for long if we may believe 
our own observations. We regard rinderpest virus as essentially fragile 
and incapable of development in external media. 


Edington(2) states: 


Similarly the nasal mucus from a spontaneous case of rinderpest was 
found to lose its virulence very quickly if exposed to the air and kept for 
any period beyond 24 hours. 


Stockman, (3) writing about the serum-alone method, observes: 


The virulent material does not remain active for more than a day or two 
outside the animal body. 

Yersin(4) states that two days of desiccation are sufficient to 
destroy the virulence of the blood. 

Ruediger(5) states that pastures which have been infected by 
sick animals may remain infected for months or even for years. 

Hutyra and Marek(6) give an extensive symposium of views 
of various writers. 

We have carried out a series of experiments bearing on the 
subject of the transmission of rinderpest designed to simulate 
natural conditions as nearly as possible. 


*To be published as Bulletin No. 30 of the Bureau of Agriculture of the 
Government of the Philippine Islands. 
* Archibald R. Ward, chief. 
124293——4 49 


50 The Philippine Journal of Science 1914 


DURATION OF RINDERPEST INFECTION IN CORRALS AFTER REMOVAL 
OF INFECTED ANIMALS 


Experiment 1.—This experiment was designed to furnish 
information regarding the length of time that rinderpest infec- 
tion would remain in a corral subsequent to the removal of 
infected animals. 

The corral had been subjected to infection by the presence of 
bull 2985, carabao 3008, and carabao 3005, each of which had 
undergone an attack of rinderpest while kept there. The period 
during which these animals collectively had displayed febrile 
temperature, diarrhea, and other symptoms of rinderpest covered 
fifteen days previous to their removal. At that time 2985 was 
in the middle stage of the attack, and had been showing a febrile 
temperature for five days. Furthermore, the corral had con- 
tained other infected cattle and carabaos during the previous 
three months. 

The corral contained a pool directly exposed to the sunlight, 
which had been used as a wallow by the carabaos mentioned 
above. There was very little shade, barely enough for the com- 
fort of the animals, and no grass. The infectiveness of the 
corral immediately after removing all animals was not checked. 
A photograph of the corral is shown in Plate I, fig. 1. For 
this experiment, the only water supply available was the above- 
mentioned pool. No one entered the corral during the experi- 
ment excepting after disinfection of the feet. The conditions 
of the experiment follow: 


February 15, 1911. All animals were removed from the corral at 8 
a.m. Sky clear and bright. 

February 16. At 8 a. m. bull 3052 was turned loose in the corral and 
left for a period of twenty-four hours. It was seen to drink from the 
carabao wallow. Sky clear and bright. 

February 17. At 8 a. m. bull 3052 was removed from the corral, dis- 
infected, and segregated along with a susceptible control animal to check 
against accidental infection subsequent to that in the corral. Sky clear and 
bright. 

February 18. Corral empty. Sky partly cloudy. 

February 19. Bull 3055 was turned into: the corral, and was seen to 
drink from the wallow. Sky partly cloudy, high wind. 

February 20. Bull 3055 was removed from corral, disinfected, and 
segregated with one control. Sky clear and bright. 

February 21. Corral empty. Sky partly cloudy. 

February 22. Bulls 3058 and 3059 were put in the corral, and drank 
from the wallow. Sky clear and bright. 


IX, B, 1 Ward et al.: Transmission of Rinderpest 51 


February 238. Bull 3058 was removed from corral, disinfected, and 
segregated with a control. Sky clear and bright. 

February 24. Bull 3059 still in corral. Sky clear and bright. 

February 25. Bull 3059 was removed from corral, disinfected, and 
segregated with a control. Sky partly cloudy. 

February 26. Corral empty. Sky cloudy and weather cool. 

February 27. Corral empty. Sky partly cloudy. Slight showers dur- 
ing night and early morning. 

February 28. Corral empty. Sky clear and very hot weather. 

March 1. Wallow was reflooded with water to replace loss from leakage 
and evaporation. Two native carabaos, 3078 and 3079, were placed in 
the corral; they were seen to drink from the wallow. Sky clear and 
bright. 

March 2. Carabaos still in the corral. Sky partly cloudy. 

March 8. Carabaos still in corral. Sky clear and bright. 

March 4. Carabaos still in corral. Rain during forenoon, clear in 
afternoon. 

March 5. Carabaos 3078 and 8079 were removed from corral, disin- 
fected, and segregated with a control. Sky clear and bright. 


None of the animals exposed in the corral in this experiment 
contracted rinderpest. The susceptibility of 3052, 3078, and 
3079 was proved subsequently by their contracting rinderpest 
after suitable exposure. Bull 3059 did not react. Record of 


3055 is not complete in this respect. 
The main details of the experiment are summarized in Table I. 


TABLE I.—Haposure of susceptible animals to supposedly infected corral. 


| Interval 
| between 


removal Ti f | 
= of sick and| ~!™¢° Susceptibility 
Animal No. | exposure pee Result. A EG. 
of sus- | 1 corral. 
ceptible 
animal. 


SOD ee ee 1 1 | Negative ___| Susceptible. 
BOGS? oh eetee Ss 4 1 eee does << Not tested. 
SODB2 sae ees 7 | iy eee dos. Susceptible. 
BOBO) aes ae 7 bo eee (eee | Immune. 
STS ae koe 14 EN ce ae WO\enc24 Susceptible. 
pal se a al 14 cl ae Batt do)... Do. 

ee 


Subsequent experiments have shown the peculiarities of the 
disease to be such that of the three animals last in the corral, 
2985 only was probably disseminating infection on February 15. 
‘This fact does not invalidate the experiment in demonstrating 
the noninfectiveness of the corral after it had been occupied for 
several previous months by infected animals. 


52 The Philippine Journal of Science 1914 


Experiment 2.—This experiment was designed to check the 
results of the previous experiment under slightly different con- 
ditions. The corral employed, designated No. 2, was well shaded 
by a tree, and contained a watering trough, but no pool. 
Infected animals had occupied this corral for nineteen days 
before the beginning of the experiment. A photograph of this 
corral is shown in Plate I, fig. 2. 

The cattle in the corral on March 7, 1911, were 3062, seventh 
day after inoculation and showing fever and inappetence; 3057, 
fourteenth day after inoculation, died same day; 3042, twenty- 
seventh day after exposure and well; 3043, twenty-eighth day 
after exposure; 3045, forty-first day after exposure and well; 
and 3048, thirtieth day after exposure, but still sick. 

Of these six, it is most likely that animals 3048, at the fifth 
day of febrile temperature; 3057, at the sixth day; and 3062, at 
the fifth day only were disseminating infection at the date they 
were removed from the corral, a fact unsuspected at the time the 
experiment was performed. All had occupied the corral for 
periods ranging from two to six days. The details concerning 
the exposure of the animals in the corral appear below. 


March 6, 1911. The following animals were removed on this day: 3062, 
3057, 3042, 3043, 3045, and 3048. Sky clear and bright. 

March 7. Bull 2999 was placed in the corral. Sky clear and bright. 

March 8. Bull 2999 was removed from the corral, disinfected, and 
segregated with a control. Sky clear and bright. 

March 9. Corral empty. Sky clear and bright. 

March 10. Bull 3000 was placed in the corral. Sky partly cloudy. 

March 11. Bull 3000 was removed, disinfected, and segregated with a 
control. Sky cloudy with slight shower. 

March 12. Corral empty. Sky clear and bright. 

March 13. Bull 3001 was placed in corral. 

March 14. Bull 3001 was removed from corral, disinfected, and segre- 
gated with a control. Sky clear and bright. 

March 15. Corral was empty. Sky clear and bright. 

March 16. Corral was empty. Sky clear and bright. 

March 17. Bull 3063 was placed in the corral. Sky clear and bright. 

March 18. Bull 3068 is still in corral. Sky partly cloudy. 

March 19. Bull 3063 was removed from the corral, disinfected, and 
segregated with a control. Sky cloudy with some rain. 


None of the cattle exposed in the corral in this experiment 
contracted rinderpest. All were subsequently shown to be sus- 
ceptible to rinderpest by contracting the disease after suitable 
exposure. 

The main details in the foregoing are summarized in Table II. 


eee eee 


IX, B, 1 Ward et al.: Transmission of Rinderpest 53 


TABLE II.— Exposure of susceptible cattle to supposedly infected corral. 


Interval 
Decgern. 
removal o: a 
* Time of Pie 
. sick and Susceptibility 
Animal No. ecposire eaveuure Result. pastiator 
of sus- 2 
ceptible | 
anim: 
Days. Days. } 
VAST tg ea 1 1 | Negative ___| Susceptible. 
S000Ss222 25-5. 4 1 eee does Do. 
SOQIN 322 224282 7 0g eee do!2= = Do. 
S068 o 2 seee === 11 Z| cee Mole Do. 


Like the preceding one, this experiment did not demonstrate 
that infection of rinderpest could persist in the corral for even 
one day after the removal of the sick. 

Experiment 3.—This experiment was designed to duplicate 
the conditions of the preceding experiments with presumably 
infected corrals, but under different weather conditions, as 
cloudy rainy weather existed during the progress of this experi- 
ment. The same corral was used as in the preceding one. 

The corral had been subjected to infection by bulls 3064, 3066, 
and 2998. On the day that they were removed the disease had 
progressed among them as follows: 3064, third day of febrile 
temperature; 3066, third day; and 2998, sixth day. The expo- 
sure of the various animals is described in the following notes: 

July 22, 1911. Bulls 2998, 3064, and 3066, all infected with rinderpest, 
were removed from the corral at 4 p. m. Weather rainy and corral very 
muddy. 

July 23. Two susceptible cattle, 3147 and 3151, were turned loose in the 
corral at 4 p.m. Weather rainy. 

July 24. Two susceptible carabaos, 3164 and 3178, were turned loose 
in the corral at 4 p.m. The cattle mentioned in the preceding paragraph 
remained in the corral. Weather rainy. 

July 25. The 2 cattle and 2 carabaos still in the corral. Sky cloudy and 
showers. 

July 26. Animals still in the corral. Sky cloudy and showers. 

July 27. All of the susceptible animals were removed from the corral 
on this date and segregated. Sky cloudy and showers. 


None of the animals used in this experiment contracted rinder- 
pest. All of them were later shown to be susceptible with the 
exception of 3151, which died as a result of exposure to bad 
weather. 

The main details of the experiment are summarized in Table 
TE: 


5A The Philippine Journal of Science 1914 


TABLE III—Exposure of susceptible animals to supposedly infected corral. 


Interval 
ela 
removal o: A 
rs Time of } +}: 
| c sick and Susceptibility 
Animal No. exposure Seppe Result. test later. 
of sus- _ 
ceptible 
animal. 
Days. Days. 
SIMI AE 1 4 | Negative ___| Susceptible. 
sible ee 1 Ae Woe. 2 Not tested. 
S168: seats: a3 2 Sizes does) Susceptible. 
S178 see ~ Ce teem does" Do. 


The chief distinction between this experiment and those pre- 
ceding lies in the rainy weather prevailing and the resulting 
muddy condition of the corral. The results coincide with those 
of the former experiments in that infection was not demonstrated 
to persist in the corral for one day. 

Experiment 4.—This experiment was essentially a duplication 
of experiment 1, except that the carabao wallow was covered on 
top and on three sides with a bamboo and grass roof. 

The corral had been infected by the presence of carabaos 3085 
and 3086, both in a stage of the disease corresponding to the 
sixth day following the initial rise of temperature. During the 
last twenty-four hours that these animals were in the corral, 
bull 2999 was kept with them as a control to show the existence 
of infection. The animal was removed, isolated, and developed 
rinderpest. During the day previous to removal, the sick cara- 
baos occupied the wallow and defecated therein. The details 
regarding the exposure of the animals in the corral appear in 
the notes below. 


April 2, 1911. Bulls 3085, 3086, and 2999 were removed from the corral. 

April 3. Bull 3104 was turned loose in the corral; it was seen to drink 
from the wallow. Sky clear and air dry. 

April 4. Bull 3104 was still in corral. Sky clear and air dry. 

April 5. Bull 3104 was removed from corral, disinfected, and segregated 
with controls against accidental infection. Carabao 3079 was put in the 
wallow on this date, being led through the corral with pads on his feet 
soaked in disinfectant. Rain occurred during the night. 

April 6. Carabao 3079 was removed from corral, and immediately dis- 
infected and segregated with control. Rain occurred during the morning. 

April 7. Carabao 3090 was put in wallow in the corral with the same 
precautions as taken with carabao 3079. Sky clear and bright. 

April 8. Carabao 3090 was removed from corral, disinfected, and iso- 
lated with control. Sky clear and air dry. 

April 9. Carabao 3087 was turned loose in the corral, and was left 
in corral one week. Sky clear and air dry. 


CO ES ee 


IX, B, 1 Ward et al.: Transmission of Rinderpest 55 


None of the animals contracted rinderpest; the susceptibility 
of all was proved subsequently. 

The chief features of the foregoing experiment are summarized 
in Table IV. 


TABLE 1V.—Eaposure of susceptible carabaos in supposedly infected corral. 


[ Interval 
Bee 
removal o: a 
a Time of naeeer 
5 sick an Susceptibility 
Animal No. exposure eer Result. Pentlater! 
of sus- , 3 
ceptible 
animal. , 
Days. Days. 
SI04=s See ss 1 2 | Negative ___| Susceptible. 
SOTO ee ea 3 il eee doj= 3.4 Do. 
BOS0s See 5 as aes doe = Do. 
SOS Te ae= oe = Tf Gleeoe domts=>. Do. 


The infectivity of the corral was tested, after intervals of from 
one to seven days after the removal of the sick, by animals kept 
therein for intervals of from one to seven days, with negative 
results. 

Experiment 5.—This experiment was planned to determine 
the duration of infectivity of virus in a corral during hot dry 
weather. Corral 3 was used for the purpose, and had been 
supposedly infected by the presence of infected bulls 3330 and 
38329. On the day that they were removed, the disease had 
progressed in 3330 to a state corresponding to the twelfth day 
after inoculation and eighth day after initial rise of temperature. 
The animal at that time showed a temperature of 40°.4 C. with 
diarrhea and inappetence. With animal 3329, the day corre- 
sponded to the twentieth after inoculation and seventeenth suc- 
ceeding the initial rise of temperature. At the time, the most 
severe stage of the disease had been passed ten days before. 
The period of exposure of the various animals in the corral is 
shown in the following notes. 


March 12, 1912. Bulls 3329 and 33380 were removed from the corral at 
1p.m. The drinking water to which they had had access was not changed. 
Further, they had intentionally been provided with an excess of Guinea 
grass, thrown on the ground in the corral so that it might become infected. 
This fodder was left undisturbed for the successors of the sick animals. 
The sky on this day was clear, and the weather hot and windy. 

March 18. At 1 p. m. bull 3341 was placed in the corral after having 
been deprived of food and water during the forenoon. The animal im- 
mediately began to eat the soiled fodder, and drank from the pail of water 
to which the sick animals had had access. The weather continued dry, 
clear, hot, and windy. 


56 The Philippine Journal of Science 1914 


March 14. At 1p. m. bull 3339 which also had been deprived of food 
and water was placed in the corral. It immediately began to drink from 
the fresh supply of water provided in the same pail as used by the sick, 
and ate fresh fodder that had been scattered over the ground. The weather 
continued unchanged. 

March 15. At 1p. m. bull 3348 was placed in the corral after having 
been deprived of food and water during the forenoon. The animal im- 
mediately began to eat fodder from the ground. Weather unchanged. 

March 16. Weather unchanged. 

March 17. At 1p. m. bulls 3341, 3339, and 3343 were removed from 
the corral and placed in stalls isolated from infection. Weather conditions 
remained unchanged. 


None of the animals developed rinderpest as a result of ex- 
posure in this corral, but all were subsequently proved to be 
susceptible. 

The principal details of the experiment appear in Table V. 


TABLE V.—Exposure of susceptible animals to supposedly infected corral. 


, | ana 
Interval | 
benwesn| 
removal o: é 
. Time of ae igs 
| Animal No. pees = esnounne Result. Buscepapity 
Sere corral. 
ceptible | 
animal. | 
| Days. Days. | 
i | 1 4 | Negative ___| Susceptible. 
3339___.___-__| 2] eee) dope Do. 
ORAS oe ees: | 4 ry (See doe .ce: ) Do. 


As usual the experiment furnished no evidence of the persis- 
tence of infection in the corral even for twenty-four hours. 

Experiment 6.—This experiment, with the exception of a 
slight variation in the weather conditions, is a duplication of 
the previous one, the same corral, No. 3, being used. Four bulls 
had been employed to infect the corral. At the time of removal, 
the disease in 3338 had reached the twelfth day after inocu- 
lation, seventh after initial temperature; 3335, sixteenth day 
after inoculation and tenth after initial rise; 3336, thirteenth 
day after inoculation and eighth after initial rise. These had 
been kept in the corral during the earlier stages of the attack, 
but 3330 was left there for only three days. When removed, 
the progress of the disease in this animal corresponded to the 
twentieth day after inoculation and sixteenth after initial rise 
of temperature. The details regarding periods of exposure in 
the corral appear in the notes below. 


IX, B,1 Ward et al.: Transmission of Rinderpest 57. 


March 20, 1912. At 2 p. m. the 4 sick animals were removed. The 
weather was cloudy, windy, and slightly cool. 

March 21. Bull 3342 was placed in the corral after having been de- 
prived of food and water during the forenoon. It immediately began to 
eat soiled fodder left by the sick, and drank from the supposedly con- 
taminated watering trough. 

March 22. Showers occurred during the forenoon and afternoon. The 
weather was hot and windy. Bull 3344 was placed in the corral at 2 p. m., 
hungry and thirsty. 

March 23. Bull 3361 was placed in the corral at 2 p. m., and imme- 
diately began eating and drinking as had the others. On this day the 
sky was clouded. 

March 28. Bulls 3342, 3344, and 3361 were removed on this date and 
placed in stalls isolated from infection. 


None of the cattle contracted rinderpest from exposure in the 
corral, but their susceptibility was proved by attack after suit- 
able exposure at a later period. The chief details of the exper- 
iment appear in Table VI. 


TABLE VI.—E«xposure of susceptible cattle to supposedly infected corral. 


Interval | 


} between 


removal of Time of | | 


Animate sick and eopanae Reanin Susceptibility 
eae nicarealt | test later. 

ceptible 

animal. 


Days. Days. 
12. 7 pari et ed ' 1 7 | Negative cel Susceptible. 


BS UE oon one 2 Go eas does | Do. 
iSii e ted 3 | Bil eee donot | Do. 
is Ae 


As usual, no evidence was produced to show that the sick 
animals left the corral infective for even a day. 

Experiment 7.—This experiment was essentially a duplication 
of the two preceding ones except that the ground in the corral 
was kept moist by sprinkling, in order to simulate conditions 
caused by wet weather. The same corral was employed as be- 
fore. Three bulls, all infected with rinderpest, had been kept 
in the corral. On the day that they were removed, the disease 
in bull 3122 had reached a stage corresponding to the eleventh 
day after inoculation and eighth after initial rise of temperature; 
in 3366, the twelfth after inoculation and seventh after tem- 
perature rise. Bull 3300 died the day before, which was the 
ninth day after inoculation and fifth after rise of temperature. 
This one had been in the corral two days; 3122, seven days; 
and 3366, eleven days, since inoculation. 


58 The Philippine Journal of Science 1914 


While the animals were in the corral, the ground was kept 
wet by sprinkling. The exposure of the animals in the corral 
is described in the following notes: 


April 25, 1912. Sick animals were removed from the corral. 

April 26. At 7.30 a. m. bull 3373 was placed in the corral. It imme- 
diately begun to eat the contaminated fodder that had been kept wet, and 
drank from the trough. At 7.30 p. m. bull 3374 was placed in the corral. 
The sky was clear, and the weather dry and windy. The ground was 
frequently sprinkled. 

April 27. At 7 a. m. bull 3375 was placed in the corral hungry and 
thirsty. The weather was hot and windy, but the ground was kept moist. 

April 28. The weather remained unchanged, and the corral was kept 
moist. 

April 29. There was a shower before sunrise, but otherwise conditions 
were the same as before. 

April 30. The 3 animals were removed from the corral and placed in 
stalls isolated from infection. 


None of the animals contracted rinderpest from exposure in 
the corral, and all were subsequently proved to be susceptible 
by contracting the disease after suitable exposure. The prin- 
cipal details of the experiment are shown in Table VII. 


TABLE VII.—LHxposure of susceptible cattle to supposedly infected corral. 


Interval 
pees 4 
removal 0; . 
Time of crac 
? lity 
Animal No. sick and exposure Result. Susceptibi 
* | exposure | ; test later. 
of sus-_ | corral. 
ceptible 
animal. 
Days. Days. 
BO ee Sone 1.0 4.0 | Negative ___| Susceptible. 
3974... ase 1.5 SS BY ee a Cs (eras Do. 
SBTD ees 2.0 Sn) aes doi -a252 Do. 


No evidence was produced to show that the sick animals left 
the corral infective for even one day. 

Experiment 8.—This experiment was similar in purpose to 
the preceding ones, and the same corral, No. 3, was used. Three 
bulls had been employed for infecting the corral. When they 
were removed, the disease in 3348 had progressed ten days from 
inoculation and six days from rise of temperature; in 3391, 
thirteen and eight days, respectively; and in 3303, seven and 
five days, respectively. The first mentioned had been in the 
corral seven days; the second, eight days; and the last, five days 


ial il lO eee ee 


IX, B, 1 Ward et al.: Transmission of Rinderpest 59 


when they were taken out. The details of exposure in the corral 
are described in the following notes: 

July 1, 1912. At 5 p. m. the sick animals were removed from the corral, 
and bull 3298 was put in the corral at 5 p. m. 

July 2. At 5 a. m. bull 3298 was removed and isolated. Bull 3402 
was placed in the corral at the same time. 

July 3. Bull 3403 was placed in the corral at 5 a. m. 

July 4. All animals were removed at 5 a. m. and placed in screened 
stalls isolated from infection. 


Bull 3298, which was used as a control to demonstrate the 
existence of infection in the corral just after the removal of the 
sick, showed a rise of temperature on the sixth day, and died 
on the twelfth day after exposure. None of the others, exposed 
later, contracted the disease, but their susceptibility was proved 
subsequently. The principal details of the experiment are shown 
in Table VIII. 


TABLE VIII.—E«posure of susceptible animals in infected corral. 


asi <<" La el ie | 
| 
Interval 
eer 
\removal o: A 
r Time of aeons 
: sick ani Susceptibility 
Animal No. | exposure | ©xposure Result. | “ ‘test later. 
of sus- | 
| ceptible | 
| animal. | 
i | 
Hours. Hours. | 
298s | None. 12 | Positive -____| 
SA0 ane ee aes | 36 48 | Negative ___| Susceptible. 
BANSAL! 6x9 48 Py ee ido. ae Do. 
= a pee 22 


No evidence was produced to show that the corral remained 
infective for over twelve hours after the sick were removed. 
The experiment in question was stronger than the preceding in 
that the existence of infection left by the sick was demonstrated. 

Experiment 9.—This experiment was similar in character to 
the last, and the same corral was used. Bull 3389 was used 
to infect the corral, where it remained during the course of the 
disease. The initial rise of temperature occurred on the fourth 
day after inoculation and death on the seventh day. The periods 
of exposure in the corral are shown in the following notes: 


June 27, 1912. Bull 3389 died and was removed at 8 a. m. The 
weather was rainy and hot. Corral was left empty until the next day. 

June 28. Bull 3395 was placed in the corral at 8 a. m. 

June 29. Bull 3395 was removed from the corral at 8 a. m. and isolated 
in a screened shed. 


60 The Philippine Journal of Science 1914 


Bull 3395, which remained in the corral for twenty-four hours 
after the corral had stood empty an equal period of time, did not 
contract the disease. It was later proved to be susceptible. 

As in previous experiments, no evidence was produced to show 
that rinderpest infection will persist in a corral for twenty-four 
hours. 

Experiment 10.—In view of the fact that failure had attended 
all attempts in the previous experiments to infect animals after 
an infected corral had been vacated for twenty-four hours, this 
experiment was performed to make tests at shorter intervals. 
The same corral was used as in the previous experiment. It 
was infected by the presence of carabao 3173, at a stage of the 
disease corresponding to the third day of febrile temperature; 
3172, fifth day; 3171, fifth day; 3160, fourth day; 3074, twenty- 
third day; 3100, fifth day; 3089, fourth day of febrile tem- 
perature. All had occupied the corral for periods varying from 
one to eight days. The movements of animals into and out 
of the corral are shown in the notes below: 


May 25, 1911. All infected animals were removed from the corral at 
4 p.m. The weather was dry and hot. At 4.30 p. m. bull 3073 was 
turned loose in this corral. 

May 26. Bull 3069 was put in the corral at 9 a. m. At 4. p. m. 2 
carabaos, 3174 and 3175, were turned loose in the corral. The weather 
was dry and hot. 

May 27. Animals still in corral. The weather was hot in the morning; 
there were showers in the afternoon. 

May 28. Animals still in corral. The weather was hot and dry. 

May 29. Animals still in corral. The weather was hot and dry with 
showers during the night. 

May 30. Animals still in corral. The weather was hot with a short 
shower just at noon. 

May 31. Animals still in corral. The weather was hot with showers 
during afternoon and evening. 

June 1. Animals were removed from the corral and placed in screened 
stalls, separated one from the other. No attendants entered the corral 
after the original infected animals were removed on May 25, and tem- 
peratures were not taken during this time. 


Bull 3073, exposed for a half-hour interval, and bull 3069, 
exposed for seventeen and one-half hours, developed rinderpest 
on the same date, showing that the corral was infective at least 
seventeen and one-half hours after the removal of the sick- 
animals. The two carabaos, 3174 and 3175, which were put in 
the corral after an interval of twenty-four hours, failed to 
develop rinderpest. Both of these carabaos contracted rinder- 
pest in subsequent experiments. 

Details of the experiment appear in Table IX. 


I Se UT 


IX, B, 1 Ward et al.: Transmission of Rinderpest 61 


TABLE I[X.—Hxposure of susceptible animals in infected corral. 


: 


Susceptibility 
test later. 


between 
removal of} Time of 
Animal No. ses anc’ | exposure Result. 
xposure | in corral 
of sus- P | 
ceptible | 
I 
| 


| Interval 


| animal. 


BO ee, 0.5 7.0 | Positive ----| 
CUTS aoe ee ae 17.5 6.0 | i eee | } 
26.5 || 
ly a pee 24.0 6.0 | Negative ___| Susceptible 
CY ee eee 24.0 ALL Seowe, doit =s.2 Do. 
* Hours. 


The results of this experiment tend to confirm those previously 
obtained. They lend color to the belief that in former exper- 
iments in this corral the infection had really perished and that 
the failure of animals to become infected was not due to mere 
accidental avoidance of infected spots. 

The series of experiments demonstrated the necessity of at- 
tacking the problem from a different angle, as described in the 
following experiments. 


PERIOD OF INFECTIVENESS DURING ATTACK 


Experiment 11.—This experiment was designed to furnish 
information regarding the relation between the period of infec- 
tiveness and the stage of the disease in cattle. It was not so 
arranged as to permit of discrimination between infection ob- 
tained directly from the sick animal on a given day and that 
which might be obtained from virus several days old that had 
resisted natural disinfecting agencies. .This latter possibility 
seemed improbable in view of the results obtained in the fore- 
going experiments. 

Two bulls, 3001 and 3060, were injected with virulent rinder- 
pest blood on April 14, 1911. These were placed in a small 
corral, 7.2 by 5.3 meters, and consequently it was possible to 
determine only facts concerning the joint infectiveness of the 
two. The corral was well shaded and usually in a muddy con- 
dition. Opportunity for the transmission of infection by close 
crowding and the use of the same drinking-water bucket and 
feeding trough were encouraged. The animals habitually placed 
their feet in the feed box. A photograph of this corral is shown 
in Plate II. The other susceptible cattle were placed in imme- 
diate contact with them in turn for a period of twenty-four 


62 The Philippine Journal of Science 1914 


hours each, after which they were removed and isolated. Im- 
mediately after removing these exposed animals, they were 
thoroughly disinfected outside of the corral. All of the cattle 
exposed to infection were afterward isolated with precautions 
to prevent accidental exposure from other sources. 

The details of the experiment appear in the double temperature 
chart of animals 3001 and 3060. The plus and minus signs 
indicate whether or not the animals subjected to infection by 
exposure or inoculation developed rinderpest when afterward 
isolated. All of these animals that developed rinderpest showed 
the initial symptoms at the usual and expected interval corre- 
sponding to the date of exposure. All of the animals that gave 
negative results upon exposure subsequently contracted rinder- 
pest in other experiments, except 3102, which did not react to 


u ey 
ie : P ANos.3001 and 306! | 
Fd tet fats fe zfs [9 [tofu fie fis [ia [15 [16 Jz ra fig {2ofarlaa]eslea [25] 26 | 
SaSe SSeS Se ee eee SEE R868 BS ee eee ee ee eee ee eee 
BERR AES Pee Se Bees Sees See See Seb Seeee feifopaferet 
BE SEER ES SSS DPB eS ee OBSalCleseo fala) 
PERSE SRSBehiesa SERS SSSR EeSe en ao 
ER SS Se SS GSH S Sioa ee Sees oeeees ae 
4.1 ERS Ss see eistetsteetert he 
ER DSSS Sess hie (ee eee eenees | naa] 
ISSN ie Se | nn 
SESS SS RSS aiaia cee Ean Deae aE fae) 
BERR BDSSS oUaxES nNeee BeesHiEe feats] Bw 
40 BERS SS SES aE ORE SNEED Peewee TJ on 
SURE ED DRE SUC SeEE li Ghee ORO an an 
BRE BRAS eaae Py TTR Tae Bw BR 
Bl 1S Be S).t ReBaee HP AAA ELE | yet ate 
3 CORR ee as fanaa) 
9 GARR Ces eo ae an 
COA AG rr ee Eo SB as 
COA Eee = are Fal an 
CeVOWER SSG Ae ay Er Ee 
C1 Ei Be AOE Ee Eee a 
38 PEE ere eo fen f pa etay 
BE SS SE SSS BS ORES Oise ESEe VERDES ENeEae ioral 
BEE EEE EEE EEE He Ss 

ele ape t | oom Spee eeeraa ce 

LU ' ‘ 


01 


s 7 Bae a ry ae ER aRED 
rat ’ F 7 1 n 
= 3 ) gs] gS} 2] 3] 8 
5 R 3 \ SPRL AL al als 


the supposedly active virus administered. Plus and minus signs 
indicate the inconclusive nature of the test on the eleventh day. 

Reference to the chart will show that an animal exposed to 
the original infected animals on the fourth day of the experi- 
ment, which was the day preceding the rise of temperature, did 
not become infected. No infection resulted on the following 
day—that of the first rise of temperature. From the sixth to 
the tenth days, inclusive, exposure in the corral, with the two 
original animals, resulted in the infection of the exposed. From 
the eleventh to the twentieth days, inclusive, when the experiment 
closed, exposure did not induce infection. Data on the eleventh 
day are inconclusive. One of the original animals died on the 
eleventh day. In the case of the animal that recovered, diar- 
rhea persisted for eight days after the last case of infection 


ee ee el a il ae 


i a i i i ee 


IX, B, 1 Ward et al.: Transmission of Rinderpest 63 


from the corral. The animals were infective during three days 
of the febrile period, and only so during two or possibly three 
days of the period when symptoms were marked. Thus, these 
two animals were infective during only two or three days of 
the period when the disease could have been recognized by in- 
spectors in the field. 

It should be noted that irrespective of the presence or absence 
of infection from 3060, no infective material, deposited in the 
previous days, remained active after the eleventh day. This 
confirms the observations in the three preceding experiments, 
and is more important because in this small crowded corral 
there was no possibility of escaping infection as animals might 
in a larger one. 

Experiment 12.—The purpose of this experiment was to deter- 


Nos. 3098 ans 3090. 


HA 
HECHEBEE 


BeSeenen 
Bare 1 
[0008 
LS 
= 


Se 
Be 
ae 
ae 
ee 
an 
an 
ae 
may 
DA 
KA 
Lim 


SREB ERESC 


Ty a ann 
Ty Baws Bea eaaa 
F 


Ae 
Cr 
1 ' 
bal Ss 
. 
Sis 
ah oy 


mine the period during which a carabao and a bull in a corral 
together would be infective to carabaos by natural exposure. 
It was a duplication of experiment 11, except as to the kind 
of animals employed. 

Bull 3098 and carabao 3090 were exposed to natural infection, 
and were placed in the same corral as used in experiment 4. 
Beginning with the eighth day after the original animals had 
contracted the disease up to the twenty-second day, excepting 
only the ninth, susceptible carabaos were exposed for twenty- 
four hours each in the corral with the carabao and bull. After 
exposure the carabaos were disinfected outside of the corral 
and isolated. The susceptibility of the animals that gave nega- 
tive results upon exposure was proved in subsequent exper- 
iments. Further details appear in the double temperature chart 
of animals 3098 and 3090. 


64 The Philippine Journal of Science 1914 


As in experiment 11, the infective period by natural exposure 
was shown to be short, and corresponded closely to the febrile 
period. Further, the infection did not remain active in the 
corral after the sick animal had passed the infective stage. 

Experiment 18.—This experiment was similar in purpose and 
technique to experiments 11 and 12, except that carabaos were 
used exclusively. Carabao 3170 was injected with virulent blood 
on June 9, 1911, and placed in the usual corral. Susceptible 
carabaos were exposed for twenty-four hours each at ten, eleven, 
fourteen, and fifteen days after inoculation, and were isolated 
as usual. The susceptibility of 3177 was demonstrated later. 
Further details appear in the following temperature chart: 


No. 370. 
rH han hima 


40 
het 


coo 
39 n 


SE ag 
385 H+-H-++ 


ai TT 


Reference to the chart will show that the animal was infective 
for at least two days preceding the appearance of diagnostic 
symptoms. The experiment also confirms the belief that the 
infective period is short and that infectious material does not 
remain active in the corral. 


INFECTIVENESS OF BLOOD DURING DISEASE 


Experiment 14.—This experiment was planned to give in- 
formation regarding the presence of rinderpest virus in the 
blood at various stages of the disease. Bull 3000 was infected 
by injection with virulent blood on April 11, 1911. Blood was 
drawn during the disease at various periods corresponding to 
five, seven, nine, eleven, thirteen, fifteen, seventeen, nineteen, 
and twenty-one days after the inoculation of the animal and 
injected into susceptible cattle. These were properly isolated 
from infection. The work is shown graphically in the temper- 


eS se oe eee 


IX, B,1 Ward et al.: Transmission of Rinderpest 65 


ature chart of bull 3000. Plus or minus signs indicate whether 
or not the animals injected with blood did or did not develop 


_rinderpest in isolation. 


The blood of 3000 was not infective on the fifth day, but was 
infective on the seventh, ninth, and eleventh days. On the 
thirteenth, fifteenth, seventeenth, nineteenth, and twenty-first 
days no infection resulted from the injection of blood from 
this animal. 

The susceptibility of the animals giving negative results in 
this experiment has been shown by their contracting rinderpest 
in subsequent experiments. 

A consideration of the results obtained from this animal, 
together with those of experiment 11, would lead to the con- 
clusion that the blood is infective early in the disease, before 


= 
> 
Go 
o 
o 
ad 
| 


& 


a 
i) 


HE HH HHH 
ERG EBebEosen 


= ae 


a 
a 
B 
a 

uy 


BERGER =S008699909030 00008 


the animal is capable of transmitting the disease by natural 
exposure. 

Experiment 15.—This experiment was similar in purpose to 
the preceding one. Blood from bull 3063 was injected into sus- 
ceptible cattle at six, eight, ten, twelve, fourteen, and sixteen 
days after the bull had been injected on April 1, 1911. The 
details of the experiment are presented in graphic form in the- 
temperature chart. 

The blood was infective on the sixth day of the experiment, 
which was the third day before a rise of temperature. It was 
likewise infective in the eighth, tenth, twelfth, fourteenth, and 
sixteenth days when the animal died. 

Observations regarding the infectiveness of blood in the later 
stages of the disease of the two animals in this and the preceding 

124293——-5 


66 The Philippine Journal of Science 1914 


experiment are not in agreement. It has been observed ® that 
blood drawn in the later stages of rinderpest did not transmit 
the disease. 


INFECTIVITY OF GRASS AFTER SPRINKLING WITH URINE FROM 
INFECTED ANIMALS 


Experiment 16.—This experiment differed from the preceding 
ones in that urine diluted with an equal amount of water was 
sprinkled on grass and animals were allowed to graze after 
various intervals. 

Five hundred cubic centimeters of urine were collected on 
April 11, 1912, from bull 3361. At 4.30 p. m. on the twelfth 
day after injection with virulent blood and the seventh day after 
initial rise of temperature, the diluted mixture was sprinkled 


a 


er 


ie) 
© 


eeC-<an 
SaeGnoR=: 


a 
a 
4 
H 
= 
z 
40H 
a 
a 
= 
wi 
HT 
it 
H 


Saag GeaRGGRe-7JEREEGROEL 


i) 
0 6) 


eae 
aan BEER EEE EES 


37H + 
Be 
3 3 
5 > 


on the grass plot, which was left unoccupied for twenty-four 
hours, the weather being dry and hot. 

On account of the expense involved, no control was used to 
demonstrate the infectiveness of the urine at the time that it 
was sprinkled on the grass. 

On April 12 at the same hour, bull 3365 was picketed over 
the infected spot and left there twelve hours to graze, after 
which it was placed in a stall. No rise of temperature was 
observed after exposure; the susceptibility of the animal was 
proved subsequently. 

In view of the absence of a control, the conclusion must be 
of a conditional character. It can only be stated that urine, 
voided at the stage of the disease indicated and left on grass for 


* Ward and Wood, Bull. P. I. Bur. Agr. (1912), No. 19, 59. 


ee ee 


ee a alent 
= ‘ 
. 


1X, B,1 Ward et al.: Transmission of Rinderpest 67 


twenty-four hours, did not convey infection to a susceptible 
animal. : 

Experiment 17.—This experiment continued the investigation 
of the results following the sprinkling of grass with the urine 
from an infected animal. A grass plot was used similar 
to that in experiment 16, and three susceptible cattle were 
employed. 

Urine was collected from two animals. Bull 3361, of the 
previous experiment, furnished part, collected on the thirteenth 
day after inoculation and eighth day of febrile temperature; 
and part came from bull 3199 on the eighth day after inoculation 
and fourth day after initial rise of temperature. 

The urine of both, amounting to 1,000 cubic centimeters, was 
mixed and diluted to 2,000 cubic centimeters in water. The 
grass plot was sprinkled with this mixture on April 12, 1912, 
at 5 p. m., and was left unoccupied for thirty-six hours. The 
weather on April 13. was cool, with showers in the forenoon, 
and was clear and hot in the afternoon. 

On April 14, at 5 p. m., bull 3366 was picketed over the grass 
plot and left there to graze until 2.30 p. m., after which it was 
placed in a screened stall. 

During the forenoon the weather was cool, with showers. 
Bull 3366 showed a rise of temperature on April 19, with 
diarrhea on April 23 and inappetence on April 25, followed 
by recovery. 

It is concluded that rinderpest virus remained virulent on the 
grass for at least thirty-six hours. 

Experiment 18.—This experiment was similar to the two 
preceding ones. 

On the sixth day after inoculation, corresponding to the third 
day of febrile temperature, 500 cubic centimeters of urine were 
collected from bull 3363 and diluted to 1,000 cubic centimeters 
with water. At 4.30 p. m., on April 16, 1912, this mixture was 
sprinkled over grass and the plot was left unoccupied for forty- 
eight hours. The weather during this interval was dry, hot, 
and windy. No control was employed to test the infectiveness 
of the urine at the time that it was sprinkled on the grass. 

On April 18, at 4.30 p. m., bull 3372 was picketed on the 
grass and left there fourteen hours to graze, after which it 
was placed in a screened stall. No rise of temperature occurred, 
but the susceptibility of the animal was demonstrated later. 

The urine of bull 3363 was tested with negative results on 
the seventh and eleventh days after inoculation in exactly the 


68 The Philippine Journal of Science 1914 


Same manner, except that the urine was exposed on grass only 
twenty-four hours. The temperature of 3363 on each of these 
dates was above 40° C. During both of these twenty-four-hour 
periods the weather was dry, hot, and windy, with cloudless 
sky. Both of the animals exposed to the grass plots failed to 
contract rinderpest, but their susceptibility was proved later. 

It is concluded that if the urine of bull 3363 was virulent 
when drawn, it was incapable of infecting a susceptible animal 
after exposure on grass for forty-eight hours in one case and 
for twenty-four hours in two instances. 

Experiment 19.—The purpose of this was similar to that 
of those experiments immediately preceding. The urine was 
voided by bulls 3368 and 3371. 

Urine from bull 3368 was collected on the sixth day after in- 
oculation—third day of fever—and from bull 3371 at the same 
stage of the attack. The total amounted to 450 cubic centi- 
meters, was diluted with water to 1,000 cubic centimeters, and 
sprinkled on the grass on May 8, 1912, at 4.45 p. m. 

The plot was left twenty-four hours, after which bull 3373 
was picketed on the ground for fourteen hours. No check was 
used for infectivity of the urine at the time of voiding. Bull 
3373 did not become infected; its susceptibility was demonstrated 
later. 

The urine of bull 3368 was tested on the seventh day after 
injection with negative results; the susceptibility of the exposed 
animal was demonstrated later. 

The mixed urine of bulls 2368 and 3371, if infective at all, 
did not remain so after twenty-four hours’ exposure on grass. 

Experiment 20.—The purpose of this experiment was identical 
with that of the preceding. 

Urine from bull 3389 was collected on the fifth day after 
inoculation, second day of febrile temperature, in the amount of 
500 cubic centimeters, and diluted with water to 1,500 cubic centi- 
meters. The mixture was sprinkled on the grass at 4.45 p. m. 
on June 25, 1912. The plot was left unoccupied for thirty-six 
hours, the weather during this time being cloudy with showers. 
- On June 27, at 4.45 a. m., bull 3394 was picketed on the plot 
and left for twelve hours, after which it was placed in a screened 
stall. The animal displayed a rise of temperature on July 2 
and diarrhcea with inappetence on July 5. Death occurred on 
July 9. 

It is concluded that the urine remained infective when spread 
on grass for thirty-six hours. 


——— SS ee ea <a 


a ee a eee 


OE Se ee eee ee 


: 
| 
! 


ES SS 


os Pe li aT ee, Be 


1X, B, 1 Ward et al.: Transmission of Rimderpest 69 


Experiment 21.—The purpose of this experiment was similar 
to that of the ones immediately preceding. The urine was voided 
by bull 3374. ' 

Urine to the amount. of 150 cubic centimeters was collected 
on the eighth day after inoculation, second day of febrile tem- 
perature. This was diluted to 1,000 cubic centimeters, and was 
sprinkled on the grass at 4.45 p.m. on June 18, 1912. The plot 
was left unoccupied for thirty-six hours, and no control was used 
for infectiveness. 

At the expiration of this period, bull 3392 was placed on the 
grass for twelve hours and did not contract the disease. The 
susceptibility of this animal was proved later. 

It is concluded that, if the urine was infective on the date 
collected, it did not remain so under the conditions afforded on 
the grass. 


INFECTIVENESS OF URINE AND FACES SPRINKLED ON GRASS 


Experiment 22.—This experiment differed slightly from the 
preceding, in that feces diluted with water were also sprinkled 
on the grass. The urine and feces were obtained from bull 3391 
on June 26, 1912, which was the eighth day after inoculation and 
third day of febrile temperature. Two hundred cubic centi- 
meters were diluted with water to 1,000 cubic centimeters and 
sprinkled upon the grass at5 p.m. The plot was left unoccupied 
for forty-eight hours, and no control was used to test the in- 
fectiveness of the excreta at the time when voided. 

After forty-eight hours bull 3396 was picketed on the spot for 
twelve hours, but did not contract the disease. Susceptibility 
of this animal was proved subsequently. 

On the twelfth day after inoculation, 500 cubic centimeters 
of feeces were diluted with water to 1,000 cubic centimeters and 
sprinkled on the grass. After twenty-four hours, during which 
time the weather was cloudy, bull 3400 was picketed on the spot; 
it failed to contract the disease. Very likely the excreta when 
voided were noninfective, due to the late stage of the disease. 

It is concluded that, if the feces and urine were infective at 
the time voided, the infection did not survive on the grass for 
twenty-four hours under the conditions existing. 


INFECTIVENESS OF FACES SPRINKLED ON GRASS 


Experiment 23.—This experiment was designed to test the 
duration of vitality of rinderpest virus in feces diluted in water - 
and spread on grass. Material was collected from bull 3448. 


70 The Philippine Journal of Science 1914 


On June 26, at 5 p. m., which was the sixth day after inocula- 
tion and fourth day of febrile temperature, 550 cubic centimeters 
of feces were collected and diluted to 1,500 cubic centimeters. 
This mixture was sprinkled on the grass which was left for 
twenty-four hours, during which time the weather was rainy. 
At the expiration of this period, bull 3392 was picketed on the 
spot for twelve hours, after which it was removed and placed 
in a screened shed. The first rise of temperature occurred on 
July 2, while diarrhea and inappetence occurred on July 5. 
Death from rinderpest occurred on July 9. 

On July 30, 1912, which was the tenth day after injection and 
eighth day after rise of temperature—and two days before 
death—600 cubic centimeters of feces were collected and diluted 
to 1,000 cubic centimeters in water. This was sprinkled on the 
grass at 4.15 p. m., after which the grass plot was left for 
twenty-four hours, during which time the sky was cloudy at 
times. Following this interval, bull 3399 was placed on the grass 
for twelve hours, after which it was isolated in a screened stall. 
No disease developed, and afterward the animal was proved to 
be susceptible. 

It is concluded that the rinderpest virus in the feces remained 
infective on the grass for twenty-four hours at an early stage 
of the disease, but not at a later stage. 


INFECTIVENESS OF FACES AND URINE AT VARIOUS STAGES 
OF AN ATTACK 


Experiment 24.—This experiment was designed to determined 
on what days feces and urine from carabaos suffering from 
rinderpest are infective to cattle. For this purpose feces and 
urine were obtained from carabao 3177 during an attack of 
rinderpest, on the fifth day after inoculation and first day of 
febrile temperature. The details of the work appear in the notes 
below: z 


August 21, 1911. Two hundred cubic centimeters of urine from 3177 
were given as a drench to 3133, which was immediately afterward isolated 
in a screened stall. 

August 22. Two hundred cubic centimeters of feces from 3177 and 
water in equal parts were given as a drench to 3117, which was imme- 
diately isolated. 

August 28. No feces nor urine were collected on this date. 

August 24. No feces nor urine were collected on this date. 

August 25. Two hundred cubic centimeters of urine from 3177 were 
given to 3157 as a drench, after which it was isolated. 

August 26. Two hundred cubic centimeters of urine from 3177 were 
given to 3121 as a drench, after which it was placed in isolation. 


; 
j 
, 
| 
f 


1X, B,1 Ward et al.: Transmission of Rinderpest val 


August 27. Two hundred cubic centimeters of feces from 3177 and 
water in equal parts were given to 3125 as a drench, after which it was 
isolated. 

August 28. Two hundred cubic centimeters of urine from 3177 were 
given to 3192, which was then isolated. 


It was decided, on account of the expense, not to attempt to 
infect more animals from 3177, but to use the results obtained 
in this experiment as a guide for future work. 


TABLE X.—Tests of infectivity of feces and urine. 


i 


Date. a ae Material. Result. Shecepiibility 
Aug. 21____ 3,133 | Urine_._.._._| Negative ___ nec al 
Aue, 225. = SRT | Mizedes: 25226 |e sae dots | 
Ce ee [atv ene Slee Sua eee 
PR Ao eins | ee ees Se 
Aug. 25____ 8,157 | Urine_-_---- Positive ___- 

| Aug. 26___- Sf A bee doje see eae dom.?-2. 
| Aug. 27__-- 8,125 | Feeces .-___- Negative -__| Susceptible. 
Aug. 28___- 8, 192) Urine = 22 — Positive soc| 


The relation existing between the stage of the disease and the 
infectivity of the feeces and urine is shown in graphic form on 
the temperature chart of 3177. Plus or minus signs indicate 
whether or not feces and urine collected on the respective days 
induced the disease in other animals. 

The disease apparently had not progressed enough on August 
21 and 22 to render infective the urine and feces tested on these 
dates, respectively. Feces were noninfective on August 27, 
while urine of the next day was infective. 


DURATION OF INFECTIVENESS OF VIRUS IN WATER 


Experiment 25.—This experiment was designed to furnish 
data as to the duration of viability of rinderpest virus in feces 
and urine when mixed with water. 

Feces and urine were collected on September 23, 1911, from 
bull 3207, on the ninth day after inoculation and the fifth day 
succeeding the initial rise of temperature. The animal had ex- 
perienced an acute attack of rinderpest, and died the same day. 
Some of the feces were gathered at autopsy. The same material 
was also collected from bull 3194 on the day of death, which was 
the eighth day since inoculation with virulent blood and the sixth 
day following the initial rise of temperature. 

A mixture of feces and urine from 3207 and 3194 was placed 
in a keg in equal parts with water, and the mixture was placed 
in the shade. 


1914 


12 


The Philippine J ournal of Science 


Two hundred cubic centimeters of this mixture were adminis- 
tered as a drench to a control animal, which was isolated in a 


screened stall. 


time. 


This animal contracted rinderpest in the usual 


Thirty-six hours later, 3209 received an equal amount of the 
diluted feeces and urine, was isolated, and developed rinderpest 


in the usual time. 


This animal 


At sixty hours, 3211 was given a similar dose. 


failed to develop rinderpest. 


On the eighty-fourth hour, 3218 received a similar dose, but 


failed to become infected. 


Bull 3215 was exposed in a similar manner at one hundred 


eight hours, and gave negative results. 


<UG0R0000000 
CCC Cee 


LETT TTT Seen eeeee 
SESEGLUEREE ESSE? wees 
HEREB SHORE R cose now seoeewe 
By ee 


74- 
2- 


cubed 
Cee se ce 
eesti aa] OT apes] 
Le 
PCC 
ee ee 708 Ht 


op) 
o 


and 3215 was proved 


’ 


The susceptibility of bulls 3211, 3213 


subsequently. 


The results are presented in Table XI. 


TABLE XI.—Tests of viability of virus in feces and urine mixed with water. 


test. 


Susceptibility 


1 
‘ 
1 


Susceptible. 
Do. 
Do. 


=] ; ev 
3 Peon ad 
a] > ee 
Cie ame wen 
a. ee at 
gf} sd 8838 
82 | gE 
a ee a a 
| =o A age ae 
1 ae Het om cr. OH 
se heat VicmRee nt 
& i ie eiaett 
< pat Nie om eat 
EEE 
033838 


= 


It is concluded that rinderpest virus from feces, mixed with 


water, survived only thirty-six hours. 


PES 1 Ward et al.: Transmission of Rinderpest 73 


INFECTIVENESS OF ANIMALS AFTER RECOVERY 


Experiment 26—The question of whether or not apparently 
recovered animals continue to spread infection afterward, like 
“bacillus carriers,” is highly important in connection with com- 
bating the disease. The assembling of 58 head of cattle that 
had been infected with rinderpest for a period varying from 
three weeks to three months previously gave opportunity to 
test the matter. Three susceptible bulls, 2998, 3053, and 3065, 
were placed among them at pasture on May 16, 1911, and 
removed June 9, after an interval of twenty-five days. They 
did not contract rinderpest, but later their susceptibility was 
proved by attack with rinderpest after suitable exposure. 

It is concluded that apparently recovered animals capable of 
spreading rinderpest by natural exposure were not present 
among the 58 head tested. 


GENERAL RESULTS BY EXPERIMENTS 


Experiment 1.—A corral which had contained many cattle and 
carabaos infected with rinderpest during the preceding three 
months and also three sick animals during the fifteen days 
preceding the removal of the sick did not prove infective to 
susceptible animals placed therein twenty-four hours after the 
removal of the infected animals. The corral was bare of vege- 
tation, was but scantily shaded, and contained a carabao wallow. 
The weather was generally clear. Six different susceptible 
animals were used to test the infectivity of the corral at various 
intervals varying from one to fourteen days after the sick were 
removed, and they each were kept in the corral for periods vary- 
ing from one to four days. 

Experiment 2.—There was employed a corral well shaded by a 
tree, but containing little or no vegetation. Infected animals 
had been in the area for nineteen days before all were removed. 
Four susceptible animals were employed to test the infectiveness 
of the corral, with negative results. Intervals of from one to 
eleven days elapsed after the removal of the sick before admitting 
the test animals, which were kept therein. The weather was 
generally clear with little rain. 

Experiment 3.—The same corral, as in the preceding exper- 
iment, was employed, but the weather was cloudy and rainy. 
Four susceptible animals failed to contract rinderpest after in- 
tervals of one and two days, after the removal of the sick ones. 
They were left in the corral for periods of three and four days. 

Experiment 4.—In this case the corral used in the first exper- 


74 The Philippine Journal of Science 1914 


iment was employed. Conditions were changed slightly by 
covering the carabao wallow on top and on three sides with a 
grass thatching. The weather was generally dry. Four test 
animals failed to contract rinderpest when exposed in the corral 
at intervals of one, three, five, and seven days after the removal 
of the sick. They were left in the corral for periods varying 
from one to seven days. 

Experiment 5.—The corral used in experiment 2 was em- 
ployed, but the weather was uniformly hot and dry. Three 
susceptible animals were used to test the infectivity of the corral 
at intervals of one, two, and three days after the removal of 
the sick, but negative results were obtained. The test animals 
were left in the corral for intervals of two, three, and four days. 

Experiment 6.—The same corral as in the preceding exper- 
iment was employed, but some rain fell during the period of the 
test. Three susceptible animals were exposed at intervals of 
one, two, and three days after removal of the sick ones, and 
negative results were obtained. They were left in the corral 
for periods of five, six, and seven days, respectively. 

Experiment 7.—The same corral was employed as in the two 
preceding experiments, but the ground was kept constantly 
moist to simulate wet weather. Three susceptible animals were 
exposed in the corral at intervals of one, one and one-half, and 
two days after the removal of the sick animals. They were left 
in the corral for four, four and one-half, and three days, respec- 
tively, but negative results were obtained. 

Experiment 8.—The tests were made in the same corral as in 
the experiments immediately preceding, and the presence of 
infection at the time the sick were removed was demonstrated 
by a control animal. This animal became infected some time 
within twelve hours after the sick animals were removed. Two 
other susceptible animals failed to contract rinderpest after 
exposure at thirty-six and forty-eight hours’ interval following 
the removal of the sick. They were left in the corral for forty- 
eight and twenty-four hours, respectively. 

Experiment 9.—The surroundings of this experiment were 
similar to those of the preceding ones. One susceptible animal 
was exposed in the corral at twenty-four hours after a sick one 
had been removed, and was left there for twenty-four hours, 
with negative results. 

Experiment 10.—This experiment was similar to the pre- 
ceding ones with regard to surroundings, but the infectiveness of 
the corral was tested at intervals of twenty-four hours or less 
after the removal of the sick animals. One animal exposed 


IX, B, 1 Ward et al.: Transmission of Rinderpest 75 


at half an hour and left in the corral for seven days contracted 
rinderpest. Another exposed at seventeen and one-half hours 
and left for six days contracted rinderpest. Two placed in the 
corral twenty-four hours after the sick animals were removed, 
and left there six days each, failed to contract rinderpest. The 
results emphasize the fact that rinderpest virus perishes in a 
bare corral in less than twenty-four hours. 

Experiment 11.—Two head of cattle infected with rinderpest 
and confined in a small inclosure were found to be infective to 
other cattle by contact on the seventh, eighth, ninth, and tenth 
days, reckoned from and including the date that they were 
injected with virulent blood. This period coincided with the 
decrease of febrile temperature regularly exhibited in cases of 
rinderpest and included the period when symptoms were most 
marked. Subsequent to the period of temperature decline, 
neither the surviving infected animal nor the infected surround- 
ings of the animal communicated the disease to susceptible ani- 
mals exposed thereto. 

The experiment is important in defining the infective period 
of a case of rinderpest and in demonstrating that virus does 
not remain infective in a corral beyond twenty-four hours. The 
results emphasize the importance of close association of animals 
in transmitting rinderpest. 

Experiment 12.—A carabao and a bull infected with rinder- 
pest and kept together in a small corral transmitted the disease 
at eight, ten, eleven, twelve, and thirteen days after they had 
been injected with virulent blood. This period corresponded to 
the stage of an attack of rinderpest, extending from the second 
day of febrile temperature until the final fall of temperature. 
It was again shown that neither the surviving animal after 
the decline of temperature nor its surroundings were capable 
of transmitting the disease to susceptible animals. The results 
confirmed those of the previous one as to the infective period 
and as to the failure of the infected corral to transmit the 
disease. 

Experiment 13.—A carabao infected with rinderpest was 
shown to be infective by contact to other susceptible carabaos 
at ten, eleven, and fourteen days after inoculation. The corral 
was not infective on the day following the death of the infected 
animal. The results verify those of the two preceding exper- 
iments. 

Experiment 14.—The blood of an animal during an attack 
of rinderpest was shown to be capable of transmitting the dis- 
ease at seven, nine, and eleven days after it had been injected 


76 The Philippine Journal of Science 1914 


with virulent blood. Tests made at five, thirteen, fifteen, seven- 
teen, nineteen, and twenty-one days gave negative results. 

Experiment 15.—The blood of an animal infected with rin- 
derpest was shown to transmit the infection at six, eight, ten, 
twelve, fourteen, and sixteen days after it had been injected with 
virulent blood. 

Experiment 16.—Urine collected from an animal on the 
seventh day of febrile temperature was diluted with an equal 
amount of water and sprinkled on a grass plot. An animal 
grazing over the grass plot twenty-four hours later failed to 
contract rinderpest. 

Experiment 17.—Urine from infected animals, at stages of 
attacks of rinderpest corresponding to the fourth and eighth days 
of febrile temperature, was diluted in water and sprinkled on 
- grass. An animal that grazed on the spot thirty-six hours later 
contracted rinderpest. 

Experiment 18.—Urine from an animal six days after in- 
oculation in the third day of febrile temperature of an attack 
of rinderpest was sprinkled on a grass plot. An animal that 
grazed thereon forty-eight hours later did not contract the dis- 
ease. Tests were made of the same urine at seven and eleven 
days after inoculation, the grass plot being left unoccupied 
twenty-four hours, but negative results were obtained. 

Experiment 19.—Urine of two animals both at a stage of an 
attack of rinderpest corresponding to the sixth day after inoc- 
ulation and third day of febrile temperature was mixed, diluted 
with water, and sprinkled on grass, but did not cause infection. 
Urine of another animal seven days after inoculation gave neg- 
ative results. 

Experiment 20.—Urine from an animal five days after in- 
oculation and two days after initial rise of temperature, when 


diluted with water: and sprinkled on grass, proved to be infec-. 


tive after thirty-six hours. 

Experiment 21.—Urine from an animal eight days after in- 
oculation, two days after initial rise of temperature, did not re- 
main infective on grass after thirty-six hours. 

Experiment 22.—Urine and feces together, from an animal 
eight days after inoculation, three days after rise of tempera- 
ture, when diluted with water and sprinkled on grass, did not 
prove infective forty-eight hours later. Feces from the same 
animal twelve days after inoculation did not prove infective 
after lying on grass for twenty-four hours. 


Experiment 23.—Feces from an animal six days after inocu-_ 


eae 


—— ee ee SS eS eee 


—e 


IX, B, 1 Ward et al.: Transmission of Rinderpest 7a 


lation, four days after rise of temperature, when diluted with 
water and sprinkled on grass, proved to be infective twenty-four 
hours later. Feces from the same animal ten days after in- 
jection, when tested in the same manner, gave negative results. 

Experiment 24.—Feces were collected from an animal during 
an attack of rinderpest at seven and twelve days after inocu- 
lation and administered to susceptible animals. The first sam- 
ple only yielded positive results. Urine collected at six days 
gave negative results, but that collected at ten, eleven, and thir- 
teen days produced the disease. 

Experiment 25.—Feces and urine were collected from an 
animal during an attack of rinderpest nine days after inocula- 
tion, five days after rise of temperature. The same material 
was collected from a case eight days after inoculation, six days 
after rise of temperature. All was diluted with equal parts of 
water in a keg in the shade. .Rinderpest was produced in ani- 
mals to which this mixture was administered, immediately, and 
after thirty-six hours. Tests made at sixty, eighty-four, and 
one hundred eight hours yielded negative results. 

Experiment 26.—Forty-eight animals that had experienced 
attacks of rinderpest within three months previously were mixed 
with 3 susceptible animals for twenty-five days without evi- 
dence being produced that they were capable of transmitting the 
disease. 
CONCLUSIONS 


1. Rinderpest virus was not shown to have survived beyond 
twenty-four hours in corrals bare of vegetation but containing 
water. The conditions under which tests were made included all 
seasons of the year with accompanying variation in sunlight, 
rain, and condition of the soil. The amount of shade varied 
widely. 

2. Animals became infected in such corrals within half an 
hour, twelve hours, and seventeen and one-half, respectively, 
after removal of the sick. 

3. Animals infected with rinderpest were shown to be capable 
of transmitting the disease to susceptible animals by close con- 
tact only during the febrile period of the disease, and most cer- 
tainly during the period in which the temperature was declining. 
The disease was not contracted by susceptible animals when ex- 
posed to sick animals during the convalescent stage when the 
temperature was nearly normal. 

4. Blood of animals infected with rinderpest was shown in 
two cases to be infected during the height of the febrile period. 


78 The Philippine Journal of Science 1914 


5. The virus in urine, diluted with water and sprinkled on 
grass, was demonstrated to survive for thirty-six hours in some 
instances, but not always, and not for a longer period of time. 

6. Feces mixed with water and sprinkled on grass infected 
an animal twenty-four hours later. 

7. Feeces and urine diluted with water and kept in a vessel in 
the shade remained infective for susceptible animals for thirty- 
six hours, but no longer. 

8. No evidence was secured to show that recovered cases trans- 
mit the disease. 

9. The foregoing facts indicate that the virus of rinderpest 
perishes soon after being discharged by the infected animal. 

10. Nothing in the foregoing experiments indicates that rin- 
derpest virus is harbored for long periods upon the soil of con- 
taminated areas. 

REFERENCES 


(1) ReFIK-BEyY and REFIK-BEY. La peste bovine en Turque. Ann. Inst. 
Pasteur (1899), 13, 596. 

(2) Epincton, A. A retrospect of the rinderpest campaign in South 
America. Lancet (1899), 1, 357. 

(3) STocKMAN, S. Report on work of veterinary department since May, 
1903. Annual Rep. Transvaal Dept. Agr. (1903-1904), 67. 

(4) Yersin, A. Etude sur quelques épizootics. Bull. économ. (1904), 
n. s. 6, 241. : 

(5) RuepicerR, E. H. Observations on cattle plague in the Philippines and 
the methods employed in combating it. Phil. Journ. Sci., Sec. B 
(1909), 4, 381. : 

(6) HutTyra und Marek. Spezielle Pathologie und Therapie der Haustiere. 
G. Fischer, Jena (1910). 


aa 
. 


ILLUSTRATIONS 


PLATE I 


iets remains alive: atts the removal of infected animals. 
a 2. Corral 2, used for determining the length of time that rinderpest 
virus remains alive after the removal of infected animals. 


PLATE II 


79 


a ea 


iia ae ity 


es sy pay . Pe 


. 


Wii fds ole Gotten oft stiniek eee 
oe hahiethphon ta poaiyt wd) ian arta badengite 
inte tes stile die Xpanek et) yriatevatahe <t Shaey cerlt 
h io 4, nial +9 snot ha? whit aria Svieabi 


Rane 


pe) ; vagine Me 


Mee rip ihe 


WARD, Woop, AND BOYNTON: RINDERPEST. |] [Pui. Journ. Scr., IX, B, No. 


Fig. 1. Corral 1, used for determining the length of time that infection of rinderpest 
remains alive after the removal of infected animals. 


Fig. 2. Corral 2, used for determining the length of time that rinderpest virus remains 
alive after the removal of infected animals. 


PLATE I. 


‘“ASV4SIG AHL 4O 3ADVLS SHL GNV SSANSAILOSSNI 30 GOlWSsd AHL NS3AMLAE ONILSIXA 
NOILVISY SHL ANINYSLAG OL SIVWINVY 40 LOVLNOOD 3S010 DNIHNOAS HOA GASN ‘€ TWHYOO ‘Il 3LWid 


“ON ‘@ ‘XI “IOS “Nunof “"1HYg] [‘LSdduaGNIY :NOLNAOG GNV ‘GOOM ‘auvVM 


INTESTINAL PARASITISM, PARTICULARLY ENTAMGBIASIS, 
IN PATIENTS OF THE PHILIPPINE GENERAL 
HOSPITAL, MANILA, P. I. 


By Davip G. WILLETS 
(From the Biological Laboratory, Bureau of Science, Manila, P. I.) 


TABLE I—Summary of findings. 


Examinations and infections. Number. | Per cent. 

Pernorns exanined. 2 3. 5 ee ee ee HS ODOT Rc. sees 
(Persons intected: oo. 2. 2 Sacco ese eeeeaun eee eases 848 84.8 

Persons infected with: 

Mareehurss: {.fe Fh ee MT eeey eS ere ee ee 606 60. 6 
SCOTIA Re Phe Be i et Be ete Se TS ee 465 46.5 
DOG TTT] 0 ea SN BN NS ah Oia es ct ee A 375 87.5 
Roolcworm)s-2 ea ea ee te eee sae eee eS se 187 18.7 
Moris) pas. gh ONS ae Ps ee Yet pe MEK es 92 9.2 
SEITE TOTO UE R Ae ee a ee See ea 13 1.3 
COREE War CAR 2 SI i a Set 10 1.0 
(OCG Trek He eta. ae oe a Se eal oh a ee 2 0.2 
Bala aiidzitiy =e. Wee We OE) PIs he Ss Fs Pe: 2 0.2 
Totatiinteetionpy: 4-225 Se. seh ed lh ee te 1, 752 175.2 


In his investigations on experimental entamcebic dysentery, 
Walker: drew, among others, the conclusions: (1) That the 
nonpathogenic F#. coli and the pathogenic E. histolytica can be 
readily differentiated by the experienced microscopist in the 
active stage, as seen in dysenteric (not diarrhceal) stools and 
in the encysted stage encountered in formed stools and (2) 
that the distinction between the pathogenic H. histolytica and 
the harmless E. coli having been established, there no longer 
exists an excuse for the indiscriminate treatment of all persons 
who show entamcebe in their stools. Granting that these de- 
ductions are valid, their practical importance rests upon two 
factors; namely, (1) the percentage of individuals harboring 
entameebe and (2) the percentage of persons infected with EF. 
coli only. 

These considerations induced me to make the examinations 
herein recorded. The objects of the work are to determine (1) 
the frequency of infection with entamcebe in patients entering 


*This Journal, Sec. B (1918), 8, 253. 
1242936 81 


82 The Philippine Journal of Science 1914 


the Philippine General Hospital and (2) the relative percentage 
of infection with H. coli and E. histolytica. Incidentally evi- 
dences of infection with intestinal parasites other than enta- 
moebee were noted. 

The helminthic findings may be compared with those obtained 
in other statistical studies of feeces in the Philippines for intes- 
tinal helminthiasis by consulting a table which was compiled 
by me.? 

A criticism which may be made of many statistical studies 
is that the conditions under which the data are secured, and 
which, therefore, limit the conclusions drawn, are not defined 
clearly. To avoid this, factors limiting the present investiga- 
tion are given as carefully as possible. 

1. The persons examined were all inside hospital patients ad- 
mitted during 1918. The vast majority of them were residents 
of Manila. Some were victims of entameebic dysentery, but 
these constituted only a small percentage of the whole number 
examined. 

2. Two thin cover-slip preparations of the one specimen ob- 
tained from each case were examined. This is obviously a 
meager examination. If more preparations had been examined 
of each specimen and particularly if repeated examinations had 
been made of negative cases, I am sure the positive percentage 
obtained would have been augmented considerably. 

3. The stool specimens were collected by the regular hospital 
routine method as follows: 

Unless otherwise specifically ordered, a patient is given a dose 
of magnesium sulphate the morning after admission and a por- 
tion of the first liquid stool obtained thereafter is sent to the 
laboratory within an hour for examination. If a patient has 
diarrhoeal or dysenteric symptoms, the first stool secured during’ 
laboratory hours is sent to the laboratory by special messenger. 
Since it was impracticable to examine all stools particularly for 
entamcebe, it was determined before a specimen was seen by the 
examiner whether it could be so examined, an operation which 
requires more time than ordinary routine examination. When 
a specimen from a patient having diarrhoeal or dysenteric symp- 
toms arrived at the laboratory, it was included in the 1,000 cases 
only provided that the next specimen, regardless of its nature, 
would have been included in the series. Thus the cases were 
unselected, but they may not represent a true average of those 
entering the hospital. 


*[bid (1911), 6, 77. 


83 


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Willets 


IX, B,1 


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84 The Philippine Journal of Science 1914 


4, Three stages of entamoebze were recognized; namely, en- 
cysted (partly or completely), active, and quiescent. No stains 
were used. The examinations were made with a Leitz 3 objec- 
tive (a Leitz 6 and an immersion lens being used when necessary) 
and a 4 ocular. 

A summary of the findings is given in Table I. The distri- 
bution of the infections according to nationality, age, and sex 
is found in Table II. 

This table shows that 375, or 37.5 per cent of the 1,000 cases 
examined, were infected with entamcebe, the highest percentage, 
46.3, being found in adult female Filipinos and the lowest, 25.0, 
in Filipino children. Experience gained at the hospital during 
the past two years inclines me to the belief that if several speci- 
mens from each case had been examined the percentage of 
persons infected would have been about one-half as high again 
as given above. In other words, instead of the positive per- 
centage being 37.5 it would have been about 55. 

The percentage of entameebic infections in inhabitants of the 
Philippine Islands recorded by other investigators and the con- 
ditions under which they were obtained are given in Table III. 

Strong,’ 1901, looking for H. coli in the stools of nondysenteric 
persons of Manila, found 8, or 4 per cent, infected in 200 exami- 
nations. Vedder,* 1906, examined 100 healthy individuals in 
the Cotabato Valley, Mindanao Province, for the presence of 
E. coli. Of 50 native scouts, mostly Moros, 35, or 70 per cent, 
and of 50 American soldiers, 25, or 50 per cent, were infected. 
Ashburn and Craig,’ 1907, examined 107 healthy American sol- 
diers at the Division Hospital, Manila, for entamebe. They 
found EL. coli in 76, or 71 per cent, and E. histolytica in 4, or 3.7 
per cent. Hoyt,’ 1908, reported 34.6 per cent infected of 300 per- 
sons examined for active entamebz at the Naval Hospital, Ca- 
facao, Cavite Province. Of 283 American sailors, 32 per cent, 
and of 17 Filipinos, 76 per cent, were positive. Garrison,’ 1908, 
in examining 4,106 inmates of Bilibid Prison, Manila, for gen- 
eral evidences of intestinal parasitism, found 23 per cent in- 
fected with active entamebe. In a similar study Garrison, 
Leynes, and Llamas,* 1909, found 2.7 per cent infected with 


* Circular on Tropical Diseases, No. 1. Manila (Feb., 1901). 
‘Journ. Am. Med. Assoc. (1906), 40, 870. 

* Milit. Surgeon (1907), 21, 348. 

* This Journal, Sec. B (1908), 3, 417. 

"Tbid. (1908), 3, 191. 

‘Ibid (1909), 4, 257. 


IX, B, 1 Willets: Intestinal Parasitism — 85 


active entamebe among 1,000 inhabitants of Taytay, Rizal 
Province. Rissler and Gomez,* 1910, report 0.39 per cent posi- 
tive for entameebe of 6,018 persons examined at Las Piiias, 
Rizal Province, for evidence of intestinal parasitism, particularly 
hookworm infection. In similar investigations the same men 
found not a single entamebic infection among 2,549 persons 
examined at Tuguegarao, Cagayan Province, and 802 examined 
at Santa Isabel, Ilagan, Isabela Province. Stitt,t° 1911, exam- 
ined 100 Filipinos of the outdoor clinic of the Naval Hospital, 
Canacao, Cavite Province, for entamcebe, finding 9 per cent 
positive. 

In the consideration of these investigations it is important to 
note several factors. In some of them fresh liquid stools ob- 
tained by the administration of a saline cathartic were examined, 
whereas in others fresh or old normal stools were used. The 
number of specimens examined of each case was not mentioned 
in a single instance. The number of cover-glass preparations 
examined of each specimen was usually not stated. The kind 
of infections sought varied considerably. -Also, the stage of 
development required for a diagnosis was not given in some 
instances, and in no case were active, quiescent, or encysted 
forms recognized. 

The data are given more concisely in Table III. 


TaBLe II].—Percentage of entamebic infections and the conditions under 
which they were obtained by various authorities. 


eee aes 
Authority and date. Exam- | Infected. Place. 
Per cent. 
iS LA ep pack CYT oe op ARM ae a a 200 4.00 | Manila. 
Me TG RENT Fer Ns 50 70.00 | Cotabato Valley, Mindanao. 
50 50. 00 Do. 
Ashburn and Craig, 1907 ___--_-----. ------ 107 71.00 | Manila. 
PRESET TE ety el aR "elt eine | 283 32.00 | Cafiacao, Cavite. | 
17 76. 00 Do. | 
Garrison: 1908/52 ne 8 4,106 | 23.00 | Manila. | 
Garrison, Leynes, and Llamas, 1909 _______ 1, 000 2.70 | Taytay, Rizal. | 
6, 018 0.39 | Las Pifias, Rizal. 
Rissler and Gomez, 1910 -____-__-__---_.--- | Dena) eetkes 20) Tuguegarao, Cagayan. | 
07-9) Snes ee Santa Isabel, Ilagan, Isabela. 
SARA O70 tee Se Se ee ea 100 | 9.00 | Cafiacao, Cavite. 


“Ibid (1910), 5, 267. * Ibid (1911), 6, 211. 


86 The Philippine Journal of Science 1914 


TABLE III.—Percentage of entamebic infections, etc—Continued. 


Species. 
Race. Sex. Age. : a Stage required for 
mh cor. a ee diagnosis. 
Per cent. | Per cent. 
(2) (2) (2) 4 (2) (2) 
Rp In OR eee ee Males -__-____-- Adult __- 70 (2) Active. Possibly 
quiescent. 
Americans! 25..-¢ 25. 35.n0"4|Uloaa CU eee Poeic (Nees 50 (2) Do. 
Doras Se ee ee doses ECO ane 71 3.7 | Active or encysted. 
D0 280 ae he ee Go 2-242." --.-do ____| undifferentiated. | Active. 
Milipinos) 3. -eactu 225-4 Acie dow 535-3 .---do ____| undifferentiated. Do. 
Mixed, mostly Filipinos ___| Mostly males___|_._..do ____| undifferentiated. Do. 
Palipings) 23-2 en ee Mixed 22ers 2--2 Mixed ---| undifferentiated. Do. 
Mostly Filipinos___._-__---|..._- o'a:2 heb Ee ..--do ____| undifferentiated. | (7) 
Filipinos. 2s-5 oooh do .f sonapinalbend do .___| undifferentiated. | (?) 
Character of specimen. 
tien a Specie) nce cen cleans ae 
Infections sought. examined cueminat aiee 
fe! ee case. hac Normal. | Fresh. Old. 
tic. 
i oe pei el St I ILS hi ¢3) (2) (2) (2) @y 75 gy 
Do)... 2225. eR ee: ee | (2) (2) Veas-0-3/5 Feet Yeses® 
Dow. 2.2 PRE ee (2) (2) NOs eeeee| [Cae ae See West=-22 
Bntamebic: ..2.25:2 asoeeno eee (2) (2) Wess] ee4sqecses Wepescce 
DO). 228 2. ak. Meee (2) (?) Weenie! sleet ees Ves: 22. 
D0 22s wae ose ee ee } (?) (2) Weak cei eeoe eee Meas 
General) ..2.: 323i ee 1 (2) West. 22. 2 ee Wesa--.- 
MOe-<-2-2 ene ee Aad NLS 8 (2) Mostly__| Some ___| Mostly__| Some. 
Hookworm 26... 2,2 eee 1 (2))7 aS. eee Messe iis seceu 4 Yes. 
Eintamosbiess-—ocu0 eee eee (?) (Ch) gta ees ee = p(y pee ot fen al Yes 


A glance at this table will suffice to convince one that the 
results obtained by the different authorities quoted cannot be 
rigidly compared, since no two of them have worked under 
precisely the same conditions with exactly the same object in 
mind. It is believed that these factors explain in a large degree 
the wide differences in the percentages of entamcebic infection 
reported; namely, from 0 to 76 per cent. 

Among the factors to be considered in making examinations 
for entamcebe not the least important is the stage of develop- 
ment upon which a diagnosis is rendered. The time is at hand 
when a diagnosis should be given not only upon the presence 
of active, but also upon that of quiescent or encysted forms. 
It is my experience that active forms only or encysted forms 
only may be present in a specimen. The quiescent forms, on the 


Ee 


le 


SS eS a ee Se a 


EE. 


he i eae ee 


IX, B,1 Willets: Intestinal Parasitism 87 


other hand, may be associated with either active or encysted 
stages—sometimes with both of them. In dysenteric stools, 
whether they be entameebic or of other variety, active forms 
are encountered almost invariably in this climate if the specimen 
be not over one hour old. In soft or in artificially produced 
diarrheeal stools, such as those obtained after a saline cathartic, 
active, quiescent, and some encysted forms are found, quiescent 
forms being the most frequent numerically. In hard, formed 
stools only encysted stages appear ordinarily. Old, liquid, or 
soft stools are not suitable for examination for the reason that 
active and quiescent entamcebe degenerate and disappear within 
a few hours. It has been my repeated experience to find such 
specimens which I had saved in the morning for purposes of 
demonstration to be absolutely negative in the afternoon. Hard, 
formed stools may be examined for encysted entamcebe any 
time within two or three days after being obtained. 

The ability to recognize active, quiescent, and encysted enta- 
meebe, after they have once been demonstrated to him, is readily 
acquired by one who is familiar with stool examinations. The 
active forms are identified by their characteristic movements. 
Walker ** has given the following description of the quiescent 
and encysted form: 


The resting entamoeba is distinguished from other bodies found in the 
stool by its size, distinctness, regularity of contour, degree of refractiveness, 
and especially by its nuclear structure. The entamcebz vary in size within 
considerable limits, but are usually from 20 to 30 microns in diameter. 
They are, therefore, larger than pus cells, or other protozoa, with the 
exception of Balantidiwm coli, that are found in the stools of man. They 
are also more refractive than pus, epithelial, or other cells found in the 
stools. The nuclear structure of the entamcebe is particularly character- 
istic. The unencysted entamceba possesses, unless in the process of 
division, only a single nucleus. This nucleus is round, or occasionally 
slightly oval or irregular, small with reference to the size of the cell, and 
appears not solid but as a refractive ring. This relatively small, ring- 
shaped nucleus appears to be absolutely diagnostic of an entameba. Only 
one other kind of cell observed in stools possesses a nucleus in any way 
resembling that of an entameba. This is an epitheloid cell, sometimes 
found in mucous stools, which has a ring-form nucleus relatively much 
larger than that of an entameba, occupying one-fourth to one-half of 
the cell. While an entameba may occasionally be observed with an ab- 
normally large nucleus, probably preparatory to division, the nucleus never 
approaches the size of the nucleus of this epitheloid cell. The latter cells 
are also less refractive and granular than entameebz. 

The encysted entameeba is round or slightly oval, more refractive than 
the resting or motile stage, and is surrounded by a more or less distinct 


“This Journal, Sec. B (1918), 8, 310. 


88 The Philippine Journal of Science 1914 


cyst wall. The nuclear structure here also is characteristic. The cyst 
contains several (from 2 to 8, depending upon the species of entameba 
and the stage of development of the cyst) ring-form nuclei usually smaller 
than, but of the same structure as, the nucleus of the motile entameba. 


I desire to add that the entamebic cyst wall is of a whitish 
color and the rest of it is almost invariably of a greenish tint 
under the 3 objective. 


DIFFERENTIAL DIAGNOSIS 


In the performance of the work herein recorded, I have 
repeatedly made provisional differential diagnosis upon quies- 
cent and active entamcebe occurring in ordinary and in 
diarrheal stools, particularly those produced artificially by mag- 
nesium sulphate. Subsequently, formed stools were obtained 
from those cases, and differential diagnosis rendered upon en- 
cysted entamcebe. The exact number of cases in which this 
procedure was adopted cannot be given, but certainly it was not 
less than fifty. The result was a thorough conviction that ordi- 
narily I, at least, cannot make a correct differential diagnosis 
upon either the quiescent or active forms in question. While 
active organisms seen in pronounced dysenteric stools can usually 
be differentiated, the one stage of development in which two 
distinct species can be discerned is the completely encysted one. 
For the detailed points of differentiation of the two species, the 
reader is referred to Walker’s article..2» Walker summarizes the 
differential points in a general way as follows: 


Motile stage. 
B. Entameba cok. 


1. Appearance porcelaneous. 

2. Refractiveness more pronounced. 
3. Movement sluggish. 
4 
5 


A. Entameba histolytica. | 
. Appearance hyaline. 
. Refractiveness more feeble. 
. Movements active in the fresh | 
stools. / 
. Nucleus more or less indistinct. | 
. Chromatin of nucleus scanty. 


one 


. Nucleus distinct. 
. Chromatin of nucleus abundant. 


ot > 


Encysted stage. 


A. Entameba histolytica. B. Entameeba coli. 


1. Cyst smaller. 1. Cyst larger. 
2. Cyst less refractive. 2. Cyst more refractive. 
3. Cyst usually contains elongated | 3. Cysts do not contain “chromidial 


refractive bodies known as | 
“chromidial bodies.” 
4. Nuclei never more than 4. 


5. Cyst wall thinner. 


bodies.” 


. Nuclei 8, occasionally more. 
. Cyst wall thicker. 


“Ibid (1913), 8, 317. 


1X, B,1 Wiullets: Intestinal Parasitism 89 


The difference in size of the cysts of the two species is very 
helpful in making a differential diagnosis. Size alone, however, 
is not dependable for the reason that small EF. coli cysts are 
found at times. Large 4-nucleated cysts have not been seen 
by me unless encystment was obviously incomplete. Since the 
nuclei lie in different plains, doubt occasionally exists as to the 
number of nuclei present in a given cyst. By using gentle 
pressure on the cover glass, the cyst wall can be broken and the 
escaped nuclei, lying just outside of the cyst wall, counted. 
Another practical point in differentiating entamecebic cysts is 
the manner in which a preparation is made for examination. 
It is my custom to make a thin preparation and to draw off the 
excess water with a piece of filter paper so that the cover glass 
fits snugly to the specimen. Thus the cysts are flattened out 
somewhat and the counting of the nuclei facilitated. This man- 
ner of making a cover-slip preparation also lessens the proba- 
bility of the cyst shifting to another field when the oil immersion 
lens, which must always be used to make a reliable differential 
diagnosis, is applied. 

Using Walker’s findings as a basis, a differential diagnosis 
was made upon cysts found in 76 cases. Completely encysted 
forms with 4 nuclei were called E. histolytica, and those with 
8 or more nuclei were called FE. coli. The results obtained are 
given in Table IV. 


TABLE 1V.—Differential diagnoses rendered upon encysted forms of 
entameba in 76 cases. 


| Species. Number. | Per cent. 
Entameba coli and E. histolytica present _____.____________-__ 45 69.2 
entomo cous only present.---—- 2-2-2 27 35.5 
Entameba histolytica only present_-_._._-_____-_--___-___--_- | 4 | 6.3 | 
PAL COUNCEDE COLPYESENG aes tae ee ae ee ee. bt Ta 72 | 94.7 
Entameba histolytica present________________________________- = 49)| 64.5 


In making the differentials, 4 cover-glass preparations were 
examined before a specimen was said to be negative for either 
E. coli or E. histolytica if cysts were plentiful; 10 cover-glass 
preparations, if cysts were scarce. In some instances a second 
specimen was examined because doubtful forms were en- 
countered in the first one. In the vast majority of the cases 
presenting a double infection, E. coli cysts greatly outnumbered 
E. histolytica cysts. 

Data with which the foregoing results would naturally be 


90 The Philippine Journal of Science 1914 


compared are furnished by Vedder * and Ashburn and Craig ** 
(compare Table III). Their differential diagnoses, however, 
are open to the criticism that they were made upon forms found 
in stools obtained by the administration of magnesium sulphate. 
It is noteworthy that Vedder recorded not only E. colt infections, 
but also other evidences of intestinal parasitism. Apparently, 
not a single infection with EF. histolytica was found, since none 
is given in his results. In as much as these investigators ex- 
amined apparently healthy individuals and my series of cases 
is composed of hospital patients, it may be advanced that one 
would expect to find a higher percentage of persons infected 
with FE. histolytica in the latter than in the former class of 
individuals. As a matter of fact only 6, or 8 per cent, of the 
76 cases upon which a differential diagnosis was rendered 
entered the hospital because of dysenteric symptoms. All of 
these were infected with both LH. coli and EF. histolytica. Five 
of them were chronic cases and one was a slightly acute case. 
The differentials, of course, were made upon cysts found in 
formed stools obtained after the subsidence of the dysenteric 
symptoms. 

It has been noted that the entamcoebe found in the evacuations 
of patients suffering from pronounced dysentery (as evidenced 
by the presence of entamcebz in great numbers in the stools, 
by therapeutic results, and by anatomical findings at autopsy, 
when the cases did not yield to treatment) conformed almost 
exclusively to the description of active FE. histolytica. In other 
words, these infections as a rule were apparently pure. Ac- 
cording to the literature on the subject, this is a common finding 
in dysenteric stools, and it seems rather remarkable when com- 
pared with the results obtained in the 76 cases in which a 
differential diagnosis was made upon encysted entamoebe. Of 
the 49 individuals infected with FL. histolytica, the infection was 
pure in only 4, or 8.2 per cent, and associated with E. coli in 
45, or 91.8 per cent. Six of these 45 cases, as already stated, 
entered the hospital because of entameebic dysentery. Most of 
the remaining 39 cases may have been carriers, but surely some 
of them were destined to develop dysentery. In other words, 
the E. histolytica infection occurring in some of these individuals 
was in the incubation period. These considerations force me 
to raise the following questions, which have occurred to me 
repeatedly. What happens to an associated E. coli infection 


% Journ. Am. Med. Assoc. (1906), 40, 870. 
* Milit. Surgeon (1907), 21, 348. 


IX, B, 1 Willets: Intestinal Parasitism 9] 


when acute dysentery develops? Is it crowded out by the rapid 
multiplication of E'. histolytica? Are there so many active LE. 
histolytica present in dysenteric stools that the relatively few 
active #. coli are overlooked even in a careful search? Does 
the sluggishly moving, porcelaneous EF’. coli take on activity and 
a hyaline appearance in dysenteric stools so that it simulates 
E. histolytica, just as the latter changes in motility and general 
appearance so that it resembles the former in nondysenteric, 
unformed stools? If the evidence obtained in numerous investi- 
gations were less strong, would not the very frequent association 
of E. coli and E. histolytica cysts in nondysenteric individuals, 
the impossibility of accurately differentiating the quiescent and 
active entamcebe found in nondysenteric stools including those 
obtained after the administration of a saline cathartic, and the 
almost exclusive occurrences of pure EL. histolytica infections in 
the stools of persons suffering from entamcebic dysentery tempt 
one to believe that, after all, there is but one species of Entameba 
in man and that its appearance varies under different circum- 


stances? These are grave questions which demand answers. 


SUMMARY 


1. Entameebic infections were found in 37.5 per cent of the 
1,000 individuals examined, in 38.8 per cent of 900 Filipinos, in 
26.0 per cent of 100 Americans, in 46.3 per cent of 417 adult 
female Filipinos, in 34.2 per cent of 383 adult male Filipinos, 
in 25.0 per cent of 100 Filipino children, in 25.4 per cent of 71 
adult male Americans, and in 27.6 per cent of 29 adult female 
Americans. Double infections were found in 59.2 per cent, pure 
E.. coli infections in 35.5 per cent, pure EL. histolytica infections 
in 5.3 per cent, FE. coli infections in 94.7 per cent, and EF. histoly- 
tica infections in 64.5 per cent. If more stools had been ex- 
amined of each patient, it is believed that the positive percentage 
would have been about 55 instead of 37.5. 

2. Percentage of entamcebe infection reported for the Phil-. 
ippine Islands by various investigators have varied from 0 to 76. 
This is believed to be explained in a large degree by the varying 
conditions under which the examinations have been made. 

3. Diagnosis of entameebic infection should be made not only 
upon the presence of active, but also upon that of quiescent or 
encysted forms. 

4. The one safe stage of development for a differential diag- 
nosis between FH. coli and E. histolytica is the encysted form 
encountered in formed stools. 


92 The Philippine Journal of Science 1914 


5. Differential diagnosis made upon encysted forms found in 
specimens from 76 individuals gave the following results: Hnta- 
meeba coli present in 94.7 per cent, #’. histolytica in 64.5 per cent, 
E. coli only in 35.5 per cent, E. histolytica only in 5.3 per cent, 
and E. coli and E. histolytica both present in 59.2 pet cent. 

6. It is probable that in dysenteric stools active FE. coli closely 
resembles active H. histolytica in motility and general appear- 
ance without a corresponding change in its nuclear structure. 


PRELIMINARY REPORT ON THE TREATMENT OF ENTAMGEBIASIS 
WITH IPECAC, EMETINE, AND NEOSALVARSAN AT THE 
PHILIPPINE GENERAL HOSPITAL, MANILA, P. I. 


By Davip G. WILLETS 
(From the Biological Laboratory, Bureau of Science, Manila, P. I.) 
With the codperation of Court R. STANLEY and PERPETUO GUTIERREZ 


(From the College of Medicine and Surgery, University of the Philippines, 
and the Philippine General Hospital, Manila, P. I.) 


INTRODUCTION 


The brillant success of Rogers(1-5) and others(6-22) in the 
treatment of entamcebic dysentery with hypodermic injections 
of emetine led us to test the efficacy of this drug as compared 
with that of ipecac in entameebiasis. The work having been 
interrupted for the present, it is deemed advisable to make a 
preliminary report of the results obtained to date. Our series 
of cases consists of 132. Of this number, 27 were dysenteric 
and 105 were nondysenteric. Eleven of the dysenteric cases 
were treated with emetine hydrochloride (prepared by Bur- 
roughs, Wellcome and Company); 16, with ipecac. The non- 
dysenteric cases were divided into 52 controls, 34 treated with 
ipecac, and 19 treated with emetine. Among the controls there 
were 8 cases of clinical syphilis with positive Wassermann re- 
actions. The administration of neosalvarsan in these cases 
seemed to have such a prompt action in freeing the intestine 
of entamcebe that they are considered separately. 

Nondysenteric cases were included in our series partly be- 
cause of the recent investigations of Walker(23) on experimental 
entamebic dysentery. He found that Hntameba histolytica 
is the essential etiologic agent of entamcebic dysentery; that 
E. histolytica and E. coli can be differentiated in the encysted 
stage by the experienced microscopist; that carriers of EL. histo- 
lytica are common; and that the incubation period in his ex- 
perimental cases varied from twenty to ninety-five days with an 


* Read before the Manila Medical Society, Dec. 1, 1913. 
93 


94 The Philippine Journal of Science 1914 


average of 64.8 days. Accepting these conclusions, it is evi- 
dent that the occurrence of entameebic dysentery in an infected 
individual can be prevented and that prophylaxis against car- 
riers of EF. histolytica can be attained by expelling the patho- 
genic entamcebe from the intestine of all infected persons. The 
procedure in such cases would be either (1) to make a differ- 
ential diagnosis between E. coli and E. histolytica and treat only 
those infected with the latter species, or (2) to treat all persons 
indiscriminately who are infected with entamebe. Positively 
to exclude EF. histolytica from an infection is difficult. In order 
to do so it would be necessary, in our opinion, that a person of 
considerable experience with entamcebe examine very carefully 
a number of cover-glass preparations of each of several formed 
stools from the infected person—an operation which would con- 
sume a great deal of time and patience. Furthermore, in the 
examination of the cysts in 76 cases, Willets* found a double 
infection in 59.2 per cent, a pure E. coli infection in 35.5 per 
cent, and a pure E. histolytica infection in 5.38 per cent. It 
follows that FE. coli was present in 94.7 per cent and E. histoly- 
tica in 64.5 per cent of the infected individuals. Therefore, 
it seems to us that the prophylaxis of entamebic dysentery by 
excluding EF. histolytica from infections is impracticable for 
routine usage in this locality. It would consequently appear 
that expelling all entamcebe from the intestinal tract of infected 
persons would be a more practicable method. In this connection 
it is necessary to note that it is commonly known that one may 
harbor an entameebic infection without the development of enta- 
meebic dysentery. Therefore, it is probable that only a small 
percentage of persons would submit to treatment in order to 


? The following case is of interest as bearing on the question of the incuba- 
tion period of entamebic dysentery. Dr. W. P. H., an American interne of 
the Philippine General Hospital, was admitted to the ward on August 12, 
1912, complaining of fever and headache. A routine stool examination 
disclosed cysts of both E. coli and E. histolytica. No treatment for the 
infection was given. On June 18, 1913, mild dysentery developed with 
streaks of blood and mucus in the evacuations. Subsequent to the sub- 
sidence of symptoms in response to ipecac and emetine treatments, HL. coli 
and E. histolytica cysts were found repeatedly in the stools. Other exam- 
inations made at irregular intervals between August 12, 1912, and June 
18, 1913, were constantly positive for entamcebe. It would appear that the 
incubation period in this case was ten months or more. The occurrence of 
a new infection between August 12, 1912, and June 18, 1913, of course, is 
not excluded, but Doctor H. states he was exceedingly careful about his 
drinking water and food during this period. 


IX, B, 1 Willets: Treatment of Entamebiasis 95 


avoid a dysentery of which there may be no danger whatsoever, 
unless the treatment be inexpensive, give good results quickly, 
and the method of administration be SIMDIe: and unattended by 
prolonged unpleasant reaction. 


THERAPEUTIC AGENTS NOW USED FOR ENTAMGBIASIS 


At the present time intestinal entameebiasis is treated with 
(1) ipecac, (2) emetine, (3) neosalvarsan, and (4) bismuth. 
Reports of each and all of these treatments deal chiefly with 
their application to the symptomatic cure of dysenteric cases, 
whereas, in keeping with our knowledge of the incubation period 
of entameebic dysentery and of the tendency of the dysenteric 
symptoms to recur, they should be applied also to the cleansing 
of the bowel of entamcebe. 

The well-known difficulties associated with the ipecac treat- 
ment prohibit it from being used extensively as a prophylactic 
agent. 

The evidence accumulated in various parts of the world con- 
vincingly proves the power of emetine hydrochloride quickly to 
relieve the dysenteric symptoms of entameebiasis; but its efficacy 
in absolutely cleansing the intestines of entamcebe is undeter- 
mined, for the reason that its effect upon entamcebe has not been 
sufficiently checked in many cases by laboratory examinations, 
as indicated in Table I. 

Allan(6), Baermann and Heinemann(7), Gaide and Mon- 
zels(14), and Marchoux(20) found that entamcebe were not 
expelled from the intestinal tract by the emetine treatment to 
the degree that the relief of clinical symptoms would lead one 
to suspect. Our results confirm their findings (see Tables VI 
and VIII). 

It may be added that, with Laveran(24), we believe a large 
share of the credit for the beneficial results obtained by the use 
of emetine in entamebiasis by whatsoever method of administra- 
tion belongs to Vedder(25) whose experimental studies induced 
Rogers(3) to use it hypodermically. 

The treatment of entameebiasis with salvarsan or neosalvarsan 
seems to offer great possibilities. Ehrlich(26) said “Hata was 
able to cure (amcebic dysentery) with a single intravenous injec- 
tion of salvarsan.” 

In his preliminary report of 12 cases of intestinal amebiasis 
and ameebic dysentery treated by the intravenous injection of 
salvarsan, Winn(27) gives the following summary: 


96 The Philippine Journal of Science 1914 


Immediately following, that is within twenty-four hours of, the intrave- 
nous injection of salvarsan we have seen the stools change from a mixture 
of pus and blood to those having a greenish bile color. 

In every case but one the number of stools per day has been reduced 
from one-third to three-fourths in twenty-four hours. 

Arsenic has been demonstrated in the stools and urine as early as five 
hours after administration and for as long as eighteen days. 

In all cases except one we have seen the amcebe disappear from the 
stools in from twenty-four to seventy-two hours. 


Winn’s work was carefully performed, numerous laboratory 
examinations for entamcebe evidently being made, but the 
exact dates are not given. The report of the subsequent his- 
tories of these patients, which is promised, will be exceedingly 
interesting. 

Milian(28) reports the rapid recovery of a case of ameebic 
dysentery in response to salvarsan. 

Wadhams and Hill(29) report 3 cases of entameebic dysen- 
tery treated with salvarsan with successful results. Their cases 
are, however, more or less indefinite. All of the patients were 
American soldiers who had contracted entameebic dysentery in 
the Philippine Islands several years previously. The disease had 
apparently become chronic in each case. The report indicates 
that the authors never saw entamcebe2 in the stools of two of the 
cases either before or after treatment and that in one of these the 
examination was made about one year after the administration 
of salvarsan. Entamcebe were present before and absent after 
treatment in the remaining case, but the number of stools ex- 
amined after treatment is not mentioned. 

The results obtained in our 8 nondysenteric cases treated with 
neosalvarsan are given in Tables V and VIII. We believe that 
these results and those of Winn are too remarkable to be ex- 
plained by mere coincidence. We are, therefore, inclined to be- 
lieve that this treatment will prove to be the most efficacious of 
the four under discussion in actually curing dysenteric and non- 
dysenteric entameebiasis. If so, a drawback to its common use 
would be the cost of this therapeutic agent. 

The bismuth, or better the bismuth-milk-saline, treatment of 
entameebic dysentery caused by E. histolytica is advocated by 
Deeks(30) of the Ancon hospital, Ancon, C. Z. This author 
states that in 60 consecutive cases he has not had a single death 
unless some complication were present and that in 190 cases 
treated by this method there has been not one relapse. It is 


also stated that in only two of all the cases treated by this | 


method were entamcebe of the species EF. histolytica found in the 


| 
. 
. 


IX, B, 1 Willets: Treatment of Entamebiasis 97 


stools later than the fourth day from the beginning of treat- 
ment. It is noteworthy that Deeks recognizes FH. histolytica and 
E. tetragena as distinct causes of dysentery and seems to accept 
James’ opinion that the bismuth-milk-saline treatment does not 
cleanse the bowel of entamoebe of the species “EH. tetragena.’’ 
As a matter of fact, it is now generally accepted that E. 
tetragena and EH. histolytica are identical. On the one hand, 
therefore, it appears that Deeks’s findings relative to the power 
of the bismuth-milk-saline treatment to free the intestinal tract 
of EF. histolytica are rather untrustworthy, while on the other 
hand the absence of relapses in his cases is extremely important. 


FACTORS EXERTING AN INFLUENCE UPON THE RESULTS OBTAINED 


Before proceeding to a discussion of our cases, it is deemed 
advisable to consider five factors which exert an influence upon 
the results obtained. 

1. Distribution of entamebe in stools.—A fact well known by 
persons familiar with stool examination is that entamcebe are 
very unequally distributed in a given specimen and in different 
specimens from an infected individual. For this reason it is 
impossible to give more than an approximate idea of the in- 
tensity of an infection and difficult to tell when a specimen is 
truly negative. Hence it is necessary to adopt some arbitrary 
standard for comparison. In this investigation a rough indica- 
tion of the intensity of infection in the first specimen examined 
was given by using +, + +, and + + +. In subsequent exa- 
minations a positive report was made as soon as it was definitely 
decided that entamcebz were present and four cover-glass prep- 
arations were examined before a given specimen was said to be 
negative. 

2. Number of consecutive negative examinations required to 
ensure one of the absence of entamebe from the intestinal 
tract.—A glance at Tables II to V will suffice to convince anyone 
that a single negative examination is quite insufficient to ensure 
one of the absence of entamcebe from the intestinal tract. In 
a general way, results obtained in this investigation give some 
indication of the number of consecutive negatives to be required. 
One negative examination was followed by a subsequent exam- 
ination in 72 instances. The subsequent examination was 
positive in 25, or 34.7 per cent, and negative in 47, or 65.3 per 
cent. The examination following 2 consecutive negatives was 
positive in 5, or 25 per cent, and negative in 15, or 75 per cent, 
of 20 cases. The following examination in 7 cases with 3 

124293——7 


98 The Philippine Journal of Science ia 


consecutive negative examinations was positive in 1, or 14.3 
per cent, and negative in 6, or 85.7 per cent. These results 
indicate that more than 3 consecutive negative examinations must 
be secured before a case may be said to be absolutely free from 
entamcebe, provided 4 cover-glass preparations be examined 
of each specimen. 

Insufficient examinations were made in a majority of our 
cases; hence, for the sake of comparing the results obtained in 
the several series of cases the arbitrary standard of two consec- 
utive final negative examinations obtained actually and by 
estimation’ (see Table VIII) is adopted. Two consecutive nega- 
tive examinations are, however, as stated above, not sufficient 
evidence of the absence of entamebe from the intestinal tract. 

3. The personal equation.—The personal equation entering 
into examinations must always be taken into consideration. It 
may be safely assumed that entamcebe, although present, were 
not discovered in some of our examinations. All of the labor- 
atory work was, however, performed by the same person (Wil- 
lets), so that the personal factor is constant in the several 
groups of cases. 

4. The effect of rest and diet upon entamebiasis—That rest 
and diet are factors which influence entamcebic dysentery is 
proved by the histories of patients entering hospitals with 
chronic entameebic dysentery. A not uncommon history of such 
a patient is the recurrence of dysentery at varying intervals 
during a period of months or years, the individual attack having 
subsided in response to rest, a modified diet, and perhaps a 
simple purgative. Since these factors influence the clinical evi- 
dences of entameebiasis, are they not to be reckoned with in 
the elimination of entamcebze from the intestinal tract? The 
results obtained in our controls, which were chiefly surgical 
cases, and hence at rest and upon a restricted diet for a part 
of the period during which they were under observation, suggest 
an affirmative answer to this question. 

5. Spontaneous symptomatic cure of entamebic dysentery.— 
While entameebic dysentery may be acute or chronic, it is most 
frequently of the chronic form, characterized by recurring dys- 
enteric attacks. We may, therefore, expect to secure appar- 
ently good clinical results in some instances with any method 
of treatment. For example, among the 133 cases here consid- 
ered, 29 entered the hospital with entamoebic dysentery, but 2, 


*The estimations were based on the percentage of cases that showed by 
actual examination two consecutive negatives. 


1X, B, 1 Willets: Treatment of Entamebiasis 99 


about 7 per cent, of them cleared up clinically before medical 
treatment began. Accordingly, these cases were placed in our 
nondysenteric series. : 
The results obtained in our several series of cases have been 
placed in accompanying tables. They will need but slight 
comment. The laboratory findings have been given in detail in 
order to emphasize the importance of securing a sufficient num- 
ber of negative examinations before a case is pronounced cured. 


DYSENTERIC CASES 


One of the dysenteric cases died. This case was treated ex- 
clusively with emetine which was administered intravenously 
excepting during the first day of treatment. Lesions of acute 
and chronic entameebic dysentery were found at autopsy. The 
patient might have been saved if emetine had been administered 
entirely by the hypodermic method; perhaps he would have died 
under any form of treatment. 

As shown in Table VI, emetine was much more effective in 
causing the disappearance of symptoms than ipecac. Of the 
emetine cases, 91.9 per cent recovered in an average of 3.6 days 
with the average administration of 0.287 gram of emetine. The 


corresponding figures for the ipecac cases are 62.5 per cent, 8.5 


days, and 16.3 grams. 

When considered from the viewpoint of expelling entamcebze 
from the intestinal tract of the symptomatically cured cases, 
the two drugs were about equally efficacious. Two consecutive 
final negatives were present (by actual occurrence and by ap- 
proximation) in 61.4 per cent of 10 recovered emetine cases and 
in 65.0 per cent of 10 recovered ipecac cases. The average 
number of days from the beginning of treatment to the first 
of the two consecutive final negatives was 8.25 in the emetine 
cases and 8.5 in the ipecac cases, and the average amount of the 
drug administered during this period was 0.484 gram of emetine 
and 16.67 grams of ipecac. 

It may be stated that the average number of days from the 
beginning of treatment to the end of observation in the cases 
discharged positive for entamcebze was 10.8 in emetine cases and 
11.6 in ipecac cases. During this period an average of 0.667 
gram of emetine and 16.0 grams of ipecac was administered. 
Failure to obtain negative laboratory results in these cases, 
therefore, was not due to observation for a lesser number of 
days or the administration of a lesser total amount of drug than 
to recover cases in either the emetine or the ipecac series. 


100 The Philippine Journal of Science 1914 


Two of the emetine cases which were discharged while still 
positive for entamcebe quickly reéntered the hospital with re- 
lapses. One of the two ipecac cases which were discharged 
unimproved was in the hospital a fraction less than five days. 
Five of the ten laboratory examinations made in the other case 
were negative. A culture was negative for dysentery bacilli 
in this case, but it is considered very probable that entamoebze 
were not entirely responsible for the dysenteric symptoms. 

The results obtained in our dysenteric cases are compared in 
Table VII with those secured by Rogers(5). It is to be noted 
that his results are more in favor of emetine, as contrasted with 
ipecac, than our own and that we secured better results than he 
did with ipecac. 

NONDYSENTERIC CASES 


By actual occurrence and by estimation, two consecutive final 
negative examinations were present in the several series of 
nondysenteric cases as follows (see Table VIII). 


Per cent. 


Control cases 11 25.0 
Ipecac cases 24 70.6 
Emetine cases if 36.8 
Neosalvarsan cases 8 100.0 


It would appear from the foregoing that 25 per cent of the 
cases apparently “cured” by ipecac, emetine, and neosalvarsan 
would have given two consecutive final negative examinations 
without treatment and hence that this percentage should be 
subtracted from cases “cured” by medication. Two factors 
are to be considered at this point, however; namely, (1) the 
number of examinations made of control cases which were 
constantly positive and (2) the average number of days from 
the beginning of observation of controls as compared with that 
from the commencement of treatment of the other cases to the 
first of the two final negatives. Table IV shows that 11, or 
61.1 per cent, of the 18 controls which were constantly positive 
were examined only twice. It is highly probable that some of 
these cases would have yielded one or more negatives if more 
examinations had been made of each case. Accordingly, 25+ 
per cent of the controls may be considered as having given two 
consecutive final negative examinations. The average number of 
days from the beginning of observation of controls as compared 
with that from the beginning of treatment of the other cases 
to the first of the two final negatives was— 


SS ee 


1x, B,1 Willets: Treatment of Entamebiasis 101 


Days. 
Control cases 11.3 
Ipecac cases 9.6 
Emetine cases 6.0 
Neosalvarsan cases 1.4 


_ It thus appears that not 25+ per cent but a lesser one should 
be subtracted from the results apparently obtained by the admin- 
istration of ipecac, emetine, and neosalvarsan in our cases. It 
also appears that the percentage to be subtracted decreases as 
we pass from the ipecac to the emetine in the neosalvarsan 
cases, becoming practically nil in the last named. 

The average total grams of drug given to “cured” cases up 
to the first of the two final negatives was— 


Ipecac 15.8 
Emetine 0.391 
Neosalvarsan 1.24 


From the foregoing it appears that neosalvarsan is the remedy 
of choice for treating nondysenteric entameebiasis and that 
emetine is inferior to ipecac in percentage of ‘“‘cures’” obtained 
but superior to it in rapidity of action. The evidence regarding 
emetine and ipecac is hence rather contradictory. Perhaps the 
explanation of this lies in the facts that the dosage of emetine 
varied widely, that this drug was administered in some cases 
hypodermically and in others intravenously, and that the ipecac 
cases were treated more persistently than the emetine cases. 
The number of cases of the emetine series treated by hypodermic 
injections and of those treated by intravenous injections is too 
small to admit of comparison. The indications were that the 
hypodermic method is the better of the two in expelling enta- 
moebe from the intestinal tract, probably because of slower 
elimination. It has the disadvantage of causing a local reaction 
at the site of injection, however, while this symptom does not 
attend cases treated intravenously. 

The average of days of treatment was greater but the average 
of total grams of drug administered was less in “uncured” 
cases than in those “cured” by ipecac or by emetine. 


SUMMARY 


1. The 132 cases of entameebiasis considered consist of 27 dysenteric and 
105 nondysenteric cases. The dysenteric cases are divided into 11 
treated with emetine and 16 with ipecac; the nondysenteric, into 44 
controls—34 treated with ipecac, 19 with emetine, and 8 with neo- 
salvarsan. 

2. A. Prophylaxis against the occurrence of entameebic dysentery in an 

infected individual and against carriers of Hntameba histolytica 


102 The Philippine Journal of Science 1914 


ba | 


consists in (1) making a differential diagnosis between E. coli and 
EH. histolytica and treating only those infected with the latter 
species or (2) treating all persons indiscriminately who are infected 
with entamebe. 

B. The exclusion of EF. histolytica from an infection is impracticable 
for routine usage because of (1) the experience required to make 
a correct differential diagnosis between LF. histolytica and E. coli, 
(2) the time required for an experienced microscopist to make 
such a differential-diagnosis, and (3) the frequency of LH. histolytica 
in this locality. 

C. Prophylactic treatment should, therefore, be confined to expelling 
entamcebe from the intestinal tract. In order that such a treat- 
ment may be widely used, it must be imexpensive, give good 
results quickly, and the method of administration must be simple 
and unattended by prolonged unpleasant reaction. 


. A. Four preparations are now in use for entamcebiasis—ipecac, emetine, 


neosalvarsan, and bismuth. Reports of each and all of these 
treatments deal chiefly with their application to the symptomatic 
cure of dysenteric cases, whereas they should be applied also to 
the cleansing of the bowel of entamebz. 

B. Results obtained by Winn in dysenteric, and by ourselves in non- 
dysenteric, entamcebiasis with salvarsan and with neosalvarsan 
indicate that this treatment may prove to be the most efficacious 
of the 4 varieties in quickly relieving the dysenteric symptoms 
and expelling entamcebe from the intestinal tract. 

A. Entameebe are very unequally distributed in a given stool and in 
different stools from an infected individual. This makes it difficult 
to estimate the intensity of an infection and to tell when a 
specimen is truly negative. 

B. More than 3 consecutive negative examinations are required before 
one may state with safety that a person is free from entamebe, 
provided 4 cover-glass preparations be examined of each specimen. 


. Rest and diet influence the clinical evidences of entamebic dysentery 


favorably, and our results in control cases tend to show that they 
are to be reckoned with in the elimination of entamcbe from the 
intestinal tract. 


. Because of the recurrent nature of entamebic dysentery, it is to be 


remembered that some apparently good clinical results will be obtained 
with any form of treatment. 


. A. In our dysenteric cases emetine gave a larger percentage of symp- 


tomatic cures and acted quicker in this class of cases than ipecac. 
B. Emetine and ipecac were about equally efficacious in expelling en- 

tamcebe from the intestinal tract. The time required to expel 

the entamcebe with the two preparations was also about equal. 


. In our nondysenteric cases neosalvarsan freed the bowel of entamcebz 


in 100 per cent of cases, ipecac in 70.6 per cent, and emetine in 
36.8 per cent. From the last two percentages an undetermined factor 
which is less than 25 per cent must be subtracted, because this 
percentage of cases would have given 2 consecutive final negative 
examinations (an arbitrary standard adopted for comparative pur- 
_ Poses) in the time limits without treatment. 


IX, B,1 Willets: Treatment of Entamebiasis 103 


iL. 


CONCLUSIONS 


The determination of the relative value of ipecac, emetine, 


and neosalvarsan in dysenteric and nondysenteric entameebiasis 
requires much further observation. 


2. 


Our work simply paves the way for additional investi- 


gations. 


(1) 
(2) 
(3) 
(4) 
(5) 
(6) 
(7) 


(8) 


(9) 
(10) 
(11) 
(12) 
(13) 
(14) 
(15) 
(16) 


(17) 
(18) 
(19) 
(20) 
(21) 
(22) 
(23) 
(24) 
(25) 
(26) 


(27) 
(28) 
(29) 
(30) 


LITERATURE REFERRED TO IN THE TEXT 


Rocers. Brit. Med. Journ. (1912), 1, 1424. 

IpEM. Ibid. (1912), 2, 405. 

IpEM. Lancet (1912), 2, 1062. 

IpEM. Indian Med. Gaz. (1912), 47, 421. 

IpEM. Therap. Gaz. (1912), III, 28, 837. 

ALLAN, Journ. Am. Med. Assoc. (1913), 60, 664. 

BAERMANN und HEINEMANN. Miinch. Med. Wochenschr. (1913), 60, 
1132, 1210. 

CHAUFFARD. Bull. et mem. Soc. Méd. Hopit., Paris (1913), III, 29, 
758, 1235. 

Dressy y MaraTTA. Sernana Médica (1913), 20, 797. 

DESTEFANO. Ibid. (1913), 20, 1189. 

DE VERTEUIL. Lancet (1913), 1, 1803. 

DopTER. Bull. et mem. Soc. Méd. Hopit., Paris (1918), III, 29, 686. 

DuFour et THrERS. Ibid. (1913), III, 29, 827. 

GAIDE et MonzEts. Bull. Soc. path. exotique (1913), 6, 491. 

HuTCcHESON. China Med. Journ. (1913), 27, 248. 

Jos et Livy. Bull et mem. Soc. Méd. Hopit., Paris (1913), III, 
29, 988. : 

La Cava. Malaria e Malatt. d. Paesi Caldi (1913), 4, 189. 

LAWSON. Brit. Med. Journ. (1912), 2, 793. 

Lyons. Journ. Am. Med. Assoc. (1918), 60, 1216. 

MaArcHoux. Bull. Soc. path. exotique (1913), 6, 313. 

MAXWELL. China Med. Journ. (1913), 27, 116. 

Roux et TRIBONBEAU. Bull Soc. path. exotique (1913), 6, 424. 

WALKER. Phil. Journ. Sci., Sec. B (1918), 8, 253. 

LAVERAN. Bull. Acad. Méd. (1913), 69, 142. 

VEDDER. Bull. Manila Med. Soc. (1911), 3, 48. 

EHRLICH. Chemotherapie. Vortrag, gehalten in Gegenwart Sr. Ma- 
jestat des Kaisers in der Hauptversammlung der Kaiser Wilhelms- 
Gesellschaft zur Forderung der Wissenschaften am 24. Oktober, 
1912. Dahlem-Berlin. 

WINN. Proc. Canal Zone Med. Assoc. (1912), 5, pt. 1, 7. 

Mm1ANn. Bull. et mem. Soc. Méd. Hopit., Paris (1913), III, 29, 626. 

WaApDHAMS and Hitt. Journ. Am. Med. Assoc. (1913), 61, 385. 

Deeks. Jbid. (1913), 60, 38. 


104 The Philippine Journal of Science 1914 


TABLE I.—Cases treated with emetine reported by different authors. 


i) i} 
Laboratory | Laboratory 
examina- Treatment. examina- Treatment. 
No. tion. No. tion. 
of Author. iat STE, of Author. 
case. case. 
Day. Re. Day.| Amount. eae ee Day.| Amount. 
| | Grains. Grains. 
1 | Rogers (2)_ Dp Sr ae | a ie ee 10 | Lyons (19) - 1| + 1 Fs 
3) + 3 3 4) — 4 & 
4 ry 6 4 
| aes doe 1 ar 1 a4 2) eee dos. il) se 2 g 
3 | Rogers (3)-| 1) + 1 5/ + 3 5 
2,— 2 13 9) + 4 t 
3 1 12 § 
rE doit oners 2 13 |, 14 § 
aj = 3 | belli, 12 |ebee 3 do -_--- 1] + | 3 i 
| 4 1 4 § 
Bula does: | 1 | a 1 A a fs De do 23 1] + 8 + 
DA 2 2 G7 — 9 § 
i al odin | Pare oferce pers oh rae 24) + 10 i 
eee doje == 1| + 1 1 16 & 
2); — 2 1 24 § 
i gel MEE (Cee 1 ar 1 1 25 § 
3|/ — 2 1 29 a 
j 3 1 30 3 
Sues doweee 1| + 1 1 31 3 
2/ - 2 1 nas [eee dass 1} + 2 § 
3 1 3 i 
Lg ee a! doe none |___-_. 1 1 4 § 
2 1 sta hs ee Coy 2225 0 ee 2 4 
4) 3 § 


8 Increased to § and then decreased to 4. > Entameebic dysentery at autopsy. 


IX, B, 1 


| No. of case. 


1 


Duration of dysentery. 


imo; 2 


3 mos __ 


10 dys - 


10 dys - 


(2) 


2 yrs___| & 


| Movements in 24 hours. 


7-14 


3-5 


10-12 


9-11 


8 


Willets: Treatment of Entamebiasis 


TABLE II.—Thirty cases treated with emetine. 


Laboratory ex- 
aminations. 


g 
3 
=) 


Result. 


1913. 
July 8 
fr 14 
July 18 
July 8 
July 14 
July 15 
July 21 
July 22 
July 23 
July 16 
\|July 17 
July 21 
July 26 
July 30 
July 21 
July 22 
July 23 | 
July 25 
July 27 
July 31 


Z 


Aug. 28 
Aug. 29 
Sept. 4 
Sept. 7 


Sept. 2 
Sept. 3 
Aug. 21 


Aug. 18 | 
Aug. 21 
Aug. 28 


Sept. 17 
Sept. 21 
Sept. 22 
Sept. 24 
Sept. 25 
Sept. 29 


shape org heh et | Peed tel tpt tte eee ee td 


uly 21-27 __--| 
| 


Date. 


July 8-12) -.--- 


uly 8-12 ____- 


June 18-25____ 
June 26-30___- 


Aug. 21-23____ 


Aug. 24-26, 30 
to Sept. 1. 


fave pA rts 


Aug. 27-30____ 
Sept. 7-12, 17, 
18, 20, 22. 


® Died. 


105 


Treatment. Result. 
| o |ope 
3 (B82 
& (oes 
i £38. 
> Shis8e 
s ag° a6 
b>) 
palesee 
5 
te F Bigs ee het ges 
s 2 215) B l> |BoR2 
® o $isa © |e |sRoD 
= A A!A B&B |A IZ 
Grams Grams: 
Hypo-|0.065| 2) 5/0.650/ 4 2 
der- 
mic 
----do ---_| 0.065 | 2] 5/0.650| 2 5 
i 
| 
1 
| 
ened cast 02044} 2 
_..-do -...| 0.065 | 2 }1o | 0.985) 2] 0 
=o do ----| 0.065 | 2/ 7) 0.910} 2 0 
| 
| | 
_.-.d0 =-=-| 05065} 2 | 3] 0.390} 4 1 
| 
..--do _..-| 0.065] 2] 6] 0.780] 5 | 
| 
----do _..-| 0.065 | 2} 4] 0.520) 3 2 
| 
£-.-do)-£--| 0.065} 2 
Intrave- | 0.022! 2 
nous. LG ESAS rail eee ee ee 
oe does.) 02 120/12 
wee dore--| 0.400) 12 


5 


106 The Philippine Journal of Science 1914 


TABLE II.—Thirty cases treated with emetine—Continued. 


| Laboratory ex- 
aminations. Treatment. 


: 
z 
+ 


covery. 


Number of consecutive 


negative laboratory 
examinations before 


Duration of dysentery. 
discharge. 


Movements in 24 hours. 


Doses per day. 


No. of case. 
| Day of symptomatic re- 


| Days. 
Total 


‘ 
3 
g 
: 


Sept. 10 
Sept. 12 
Sept. 16 
Sept. 17 
Sept. 27 


‘Sept. 13 

Sept. 16 

'|Sept. 17 

10 | 4 mos--; 4-5 Sept. 18 
| 


fo 3) 
nan 


9 | 5dys _- 5 


Sept. 14, 20 ---|_-_. do __..| 0.022 
Sept. 15-18, 22_|___-do ___.| 0.044 


i) 
—— 
a 

J 
wo 
& 
oo 
—] 


11 | 15mos_| 12 2) 2) 0.128) 3 1 


oe. A-5i ete Intrave- | 0.032 
nous. 


ept. 9-11 ____| Intrave- | 0.022; 2) 3) 0.182 
nous. 
- i-18 se do -_.-| 0.065 


13 0 Oo | - 
er- 


mic. 


er ----| Hy po- | 0.065 | 2| 4 | 0.520 }___. 3 


14 0 0 Suly 14-26 -_--|____ do -_--| 0.065 | 2 | 18 | 1.690 |___- 0 


tet eet TL Pit di et +eee et eee brett ba 4 


> 
5 


a 
Ee 
< 

+ 
+ 


ee 

> 

=] 

=9 
SSrekss 
ij t+4++ 

oe 

J 

z 

8 

2 

° 

i—) 


.---do -..-| 0.065 | 2) 4 | 0.620 |.-_. 0 


16 0 0 


1] 0 0 


qu 
ij 
Fe 
< 
8 
i 
i 
H 
' 
H 
1 
t 
a 
C) 
1 
1 
{ 
' 
° 
re ) 


|: 0.176 |---- 


" 
wae 


IX, B, 1 Willets: Treatment of Entameebiasis 


TABLE II.—Thirty cases treated with emetine—Continued. 


paaentderredti Treatment. | Result. 
|) a es | | eee aa caer = * 
B rls | 4 [epee 
Stl 8 | 2 1339 
ae z 1354 
ae a] r £5528 , 
eves) 3 Ihe Esisgae 
$| & 5 < Py Renee g 
‘3 S Eg A = i) a - = |3 |a & gS 
A g > 2 = 2 rr v o| 2 o Box.2 
6 5 $ 3 3 a 3 2/8/81 8 |8 |paou 
Zi a b= A fe Q = a Vata =e ja la 
1913. ' Grams} Grams 
July 23} + 
17 0 0 {ra 30 4 fren. Hypo-/| 0.044) 2); 2} 0.176 |__-. 0 
Aug. 1 Fic al dermic. 
July 26| + | July 26_-____- _.--do ----| 0.044} 2 
18 0 0 [ase 4) — | duly 27-.....|._do-...| 0.044 | 2 ]7 8 | 0.806 |___. 0 
Aug.14| + | July 28-___- _---do ....| 0.065 | 2 
July 29 ae 
' 0 0 ne ce ic 30-31; 
F Aupiles 8 PS does. 0s06D8|n2 |), 31] 0; 390) |---| 2 
Aug. 4 _ | 
Aug. 4) ++ 
Aug. 19} — | 
20|/ 0 Mapes ee Nea alta 16-18-21 4 do ....|0.065| 2} 3]o0.s90|...| 3 
Aug. 22| — 
\|Aug. 25 _ 
; ||Aug. 29 _ 
\fAug. 23 Re 
21 0 0 fen 9 + fs 10-12____| Intrave- | 0.022} 1] 3 | 0.066 |____| 0 
\(Sept.17) + nous. 
Aug.26} + | 
Aug. 29 + | 
Sept. 6 =a 
22 0 0 Sept.13 | + ept. 12-18__.-|___.do -_..| 0.022} 1] 2 | 0.044 |___.| 0 
Sept. 18 —_ | | 
Sept.19| — 
Sept.26) + | 
23| 0 0 ee an Sa \sevt Age aot! Intrave- | 0.022| 2| 1] 0.044|..| 1 
Sept. 13 = | 
nous. | } 
Sept. 6; + 
2%} 0 0 ae ~ |fSept. 8-11 -...|____do | 0.022] 2| 4] 0.176 |...) 4 
Sept. 16 _ 
Sept. 17 — | 
Sept. 9| + | 
Sept. 13 + 
25; 0 CSS oe eam oper Yay (ae do | 0.022! 2| 2/0.088|....| 4 
Sept. 17 = F 
Sept. 20 — 
Sept. 23 = 
2%| 0 0 fee ? |sevt. fis etal eed ---|'0.028 | 2) 2 | 0/088 |---|, «0 
7: 0 ue : \sevt. 18-19 ___|____.do-.| 0.022 | 2| 2 | 0.088 |... : 


108 The Philippine Journal of Science 1914 


TABLE II.—Thirty cases treated with emetine—Continued. 


Laboratory ex- Treatment. : Result. | 


aminations. 
: o ‘ 2 >o 
aed ba a a 
~ 2 = S 3 
aes @ (gs 
= Nn 4 = . ze 2 
‘) = > SPESE 
. a Sy = ao 8 = o 
= ° s Bi oe & 
3 = & 3 Bs os 
oO ° i) : He a Des F] 
4 = & 3 3 | : . w (33 ES 
So s q 3 3 ; = é 3\/ a] gd |° \ERse 
Sie 818 | week ae = So} B || a| & iB ease 
a (a) = QA io is) = A |A/A| & {A 14 
| 1913. | pal Grams 
Sept. 19 =--2-2 Hypo-/ 0.022] 2 
98 0 0 (aang 1) + dermic. 3 | 0.252 |___ 0 
|(Sept.80| + Sentiy20) <.. Sesee do --_| 0.044 | 2 
(01: Eee ye do ___| 0.065 | 2 
Sept. 22 | ++ 
Sept. 23 | + | 
Oct. 1 oF | | Y 
29 0 0 Oct. 2 = Sept. 24-27 ___| Intrave- | 0.0832 | 2) 4 | 0.256 |___- 0 
Oct. 4 ae ; nous. 
Oct. 12) + | 
Oct. 14 | + | | 
Sept.30) ++ ls 
Oct. 2) + | 
Io} 0 | 0 Kock 7| — |foct.4,6,6,12..|.....do...| 0.082 2] 4]0.256|.| 0 | 
Oct. 10 | + | 
Oct. 12 | + . 
| 
TABLE III.—Fifty cases treated with ipecac. 
zz ee = Tl = ae 
Posectnbsir Treatment. Result. 
— a a aa. een iho,” ieerbes ae 2 (oa 
s | 2 | S| eeee 
= | = FS O35 9 
5 S ares o§ OO 
a | a . “=| §o8 F 
> co a ("A OSs Eas 
Bie eS E Be | wees 
9 6 Ba Fy ao Seka 
a ec £ to cs) n ve 
° o i A S Qo = 2 
om 3 E 3 & “ i ° go re 
° oc On 0] 3 ° o @ i) > 
Bey ST) epi eg 3 Se oe 2 gesa 
a i=} = Q & i=) i=) i=) B =} 
1912-1913. Grams. 
; Aug. 6 aa ; 
it (2) ‘| 610 %Aug.10| + |pAug.8,9, 11-19.) 2.0} 11 22.0 12 1 
Aug. 16 = 
Aug. 14| ++ 
2 (2) 8-9 Aug.21| + |fAug. 15-27____- 2:0), 18 26.0 | improved. 1 
Aug. 26 = 
8 | 2 yrs... 4me"7 ee Si), Ft Isept (108-22 2.0| 7] 14.0 5 1 
7 | Sept.11 | — 
als mos... 10 foe Sly oe loet. 870: a ae 2.0! 7| 14.0| improved. 1 
Oc) 


on 


oo eee 


10 


il 


13 


| No. of case. 


Duration of dysentery. 


2 mos--- 


Willets: Treatment of Entamebiasis 


Movements in 24 hours. 


7-10 


Laboratory 
examinations. 


& 
a 


1912-1913. 
Oct. 8 
Oct. 11 


Oct. 


Treatment. 

a 

S 

a 

I 

cy) 

a 

$ 2 

3 i) 

i=} [=] 
os S16252 2s) 2.0 
Inv. 27-Dec. 2_| 2.0 
Dec. 20-Jan. 2, | 2.0 

5-9 
Jan. 28-Feb.8, | 3.0 
21-26 

ie B=13)2-5254 2.0 
br 26-Mar. 4_' 2.0 
his. Ce Gyanaeee 0.6 
be 18-27/2-= = 0.3 
2.0 


TABLE III].—Fufty cases treated with ipecac—Continued. 


Result. 
oS ita 
r=] 0 26 % 
2 BSER 
a. O93 Ss 
Eg BES 
>S SaKe 
aio ov 
% Be 8 
é| ¢ % gore 
> > 282 
as 3 BPHra 
a| = a Zz 
Grams. 
9 18.0 9 1 
1 
6 12.0 | unimproved. 74 
19 38.0 improved. 1 
18 54.0 16 8 
6 12.0 improved. 2 
7 14.0 5 if 
7 4,2 3 2 
| 
10 3.0 10 2 
11 22.0 9 1 


110 The Philippine Journal of Science 1914 
—Fi cases treated with ipecac—Continued. 
TABLE III.—Fifty treated with ip Cont d 
pee eS 
i Laboratory 
examinations. Treatment. Result. 
: =| 
> x | ° foo 
Egle g |ge8. 
c = £ aay 4 
2 a gs Rass 
> ~ di 5 SOEs 
so & £ £ o 13 a5 
© - a a BH sSsa 
QO ° » “a a2 Oo X:= 
s c c S 3) & oO 
oS i) ° . c bs Gy g 2 
% 3 5 : = : rade ae tl eee © Sons 
ee 8 ee ay eee 5 zi/2@| 3 z | 5588 
Z a = a a fa) Aiea | oS a Z | 
1912-1913. Grams. 
+ ! 
a. i 
' 
ul} + |\Mar.28-Apr.2,| 2.0} 15] 30.0 |unimproved.| ~~ 2 
+ |{ 11-19 
+ | 
ce | ; 
| eu 
ae 
15 '3 wks____! ++4 ae 18-26. ___- 2.0| 91 180 5 1 
a ; 
16 (2) — |pApr. 28-May2-| 2.0| 5] 10.0 1 2 
/ =: 
VW 0 _ Sept. 11-16____- 2.0 6 iba}, Seeaeeeenee tee 2 
ee 
— | 
18 0 — '}Apr: . 25-80)"\210" | 2] ar) ae ee vl 
me rag Oct. 9-14. | 
i 
Ley 
+ / 
19 0 s. loet. $10.2 | 20| 8| 16.0 ppsseoeceeeeaes 1 
B | | 
20; 0 “ |}Oct. 12-15_-_--- 2.0] 4 | 2) Rese ze 1 
i i 
21 0 | i |nov 6:20 aL | OI Ii! BpsOy ee 2 oS 1 
; 
22) 0 #3 Inov. 14-21... 3.0) 8| 24.0 | Paete OE o' 1 
a 
+ j | 
23 0 +  |bNov. 16-26_..__- | Bo} 10 ]* Gro} Pee ee | 2 
| ! 
/ e | 
24 0 uf Nov. 20-22 Te Rone 3.0 3 Ry | ees See 1 
+ 
25 0 |nov. 7 Dec.8..| 2:01) Siti a0 |S 1 


IX, B,1 


| No. of case. 


29 


30 


$1 


82 


34 


Duration of dysentery. 


i) 


i) 


Willets: Treatment of Entamebiasis 


TABLE III. —F%fty cases treated with ipecac—Continued. 


Movements in 24 hours. 


Laboratory 
examinations. 


111 


Treatment. 


Result. 


Date. 
Result. 


Date. 


Dose in grams. 


Days. 


Total. 


cure. 


‘ 


Day of symptomatic 


1912-1913. 
5 


Feb. 
Feb. 11 
Feb. 
Feb. 
Feb. 6 
Feb. 
Feb. 
Feb. 
Mar. 11 
= 19 


Apr. 1 
Apr. 


Apr. 1 
Apr. 8 
Apr. 12 


Apr. 16 
Apr. 18 


Apr. 2 
Apr. 9 
Apr. 14 


Apr. 23 
May 5 
Apr. 9 
Apr. 11 
Apr. 15 


Apr. 21 
Apr. ll 


: a + + ++ 
+ 
[Piero sie teterintinet eatin lanl ctiete eterna I tte bE 


Feb. 7,8, 10, 11, 
13, 15, 16. 


Mar. 20-28 ____- 


o phe ee 


4, 6-22 ____ 


Feb. 6,7, 18, 21, 
24, 28. 


Apr. ‘ 12-15, 
18-20. 


7 14-19) oes 


|. 16-22, 25_- 


Apr. 20-26, 
May 2-6, 12-16 


3.0 


2.0 


2.0 


2.0 


2.0 


2.0 


2.0 


2.0 


2.0 


10 


18 


7 


18.0 


20.0 | 


36.0 


14.0 


16.0 


34.0 


tive negative labora- 
tory examinations 


Number of consecu- 
before discharge. 


112 The Philippine Journal of Science 1914 


TABLE IJ1.—Fifty cases treated with ipecac—Continued. 


Laboratory 


| examinations. Treatment. Result. 


cure. 
tory examinations 


tive negative labora- 
before discharge. 


a 
g 
a 
=) 


a 1912-1913. Grams. 


Apr. 22 + 
Apr. 24 + 
Apr. 28 + 
35 0 0 May 5} + 


Day of symptomatic 
Number of consecu- 


Duration of dysentery. 
Movements in 24 hours. 
o 


No. of case. 
Result. 


Days 
Total. 


Date. 
| Dose in grams. 


Apr. 22, 23, 29, | 2.0 8 WGRON EX es 255. 3 
30, May 6-8. 


= to 
36 0 OMe as | Apr. 28-May 11. BG Gel ae eee eet 1 


37 0 o \JMay 1 


a 28-May 4.| 2.0 7 ERO) [oe cee Sess) 2 


88 0 0 prs (295- S06| 20H) 06 NW says eae 2 


39 0 0 May 2-6____--_- POs a5 LOONce eee eee 1 


40 0 0 May 5-9______-- Digi Ba. Wo eee sees 2 | 


41 D } 0 


43 0 0 May 17-20__---- 2.0 4 B20 ee ee eee 2 


5 
< 
8 

| 


{ 

! 

k 
i, 
May 21) + /|'May 22-26_.._- BO Bil MORON On nee 2 d 
|sJune Crib eee B0;| (Gil BNO He ase Se 1 


45 0 0 | 


June 4) ++ 
46 0 0 June 9) + une 6-10___-- 3.0 5 LES it eee oe eee 1 


IX, B, 1 


Willets: Treatment of Entameebiasis 


TABLE II].—Fifty cases treated with ipecac—Continued. 


| | 
Laboratory 
examinations. | Treatment. / Result. | 
: Bel fa re l4sa2 
Be we | wer hoe: we Bens 
c= = | ; Vor gy 
=] =] 22e 
5 = | 6 ea ah 
2 a : = Sona 
ee sca | a Gea TOY ed r= A 2 
Biel ES 2 H 8 | > Oo = 
SB | ae le ats Be ag | ne. 9 
= . E apes & ree ot Bo Beak 
3 o) 5 3 a © a | | & Cue 
ok SA ea Z cs EES ee a ne a oe 
Z fa) = a] ce a a ahs a Z 
1912-1913. Grams. 
{Aug.18| + | aes | | 
| (aid vasa 4.0 2) 0 
47 0 0 jlaug.22|/ + | | 8.0 |----------_--- 
Aug. 26; + | 
48] 0 0 ySept. 1) — jane. 26-29.....| 20) 4) 80) be ae: 
Sept. 5) — | | | 
| Sept. 6) ++) | 
49 0 ae Sept.13 | — (Sept. 11-13_....| 2.0 4 BOu ees es ea | ty) 
_ (sept. 15 | + | | 
'{Sept. 21 | sree : | 
50; 0 0 Isept.25 & |sept. Le ieee | mall | Sir ie a | 
1 i a IE 3 J} 
TABLE IV.—Control cases. 
lo = | i) | ies ae | 
| Laboratory examinations. j Number | Laboratory examinations. | Number 
2s Sa of con- [eee Ss | of con- 
| secutive | | | secutive 
No. | | | negative || No. | negative 
of | | | laboratory | of | laboratory 
ee Date. | Result. | exeniia- | ein Date. Result. | area 
| | before dis- | ; | before dis- 
i | | charge. || . | | charge. 
+ nl Er i j AO eS ne gee 1 
| | | 
Ss ay ides. N | + il fe 9 aoa | + \ F 
Reb. NB 228eos. 2 hosing | i (Aug. 7 Bees sone: Bae ity 4] 
ENDE Qieeessssea-es ta ak | PU Gap cle fs i} 0 
Cosel WEN Daa Ee eee + 4 ieee Septet = + | 
aiey sae Ie et - Horus eee | + | 
MUS ee eee == | 10 aug. 12 eee ao 8 ae 0 
Seve hale ee ~ f 1 fal VT fem a a Dart. & 
Ct Cye7 tee een me ae = (pAmies19) 2. sae sr | 
ores ie San nea chan | | LS) eee ae so | 
age xB - | Sst eee oy Sea — | 0 
4 PAGpe Id, = Fo ot oo = i 0 | ent 4s. 50 5 | 
<a Groh ae Se ~ | | TASC, (ees Sete ee + 
Vee Ca a am i].12.) Aug. 20... - 2 
ng 26 a | | meet ee = 
ulyalo 2-25 + | | Aig 'G = 83 + 
ages i fee 
5_|[Suly 21 | A 13 Ave. 9 ; 
(ia) Ceara Se + | Vr ey 1 ae See ee te | 
SCA a eel + / (Aa 019) =e so + 
ee ee ae | = weep rsosie + 
3 fee 15 + | a hogee ee 8 } 0 
SbOLYy) 26.235 een Se Arie 2 ae oS zr 
aly? 24 ooo 22s. =o ae i [aoe Biren oom of 1 
edule: 26 22/255 = 0 rl ede ~ 
alg See a! 
124293——8 


@ 


114 The Philippine Journal of Science 1914 
TABLE IV.—Control cases—Continued. 
Laboratory examinations. ember Laboratory examinations. Number 
mecntive | aaa 
No. negative || No. negative 
of laboratory || of laboratory 
Resch Date. Result. oho. CBE: Date. Result. Seong 
before dis- before dis- 
charge. charge. | 
Ao 12-322. es + YY ee ae ee + | 
16__ fave. Va Mi Mie ete — Q || 28-- fs Gyo eee - 2 
Avie Ob) eed ees i iSept. 23: ea _ 
11... lake —— A we lievq pe ipcrnreeeG ae + 
AUP); 20) 2 -cs See = et e rig eee ee | Sons 1 
(Aug18). c=. + Septem ss oe + 
AUR O Th eae ar see Net. 4 eee + o 
Ag \28 i222 ee ee — Sept.w3'--2-20 ee 4 } 
aS Sept. 52). 2.8020 + 0 || 81_- ‘eae |r Weed + i 9 
Sent.9) 222 oeeeeee sF Sept 4, en eee + 
Sept + | 32_ i Thy lh i _ | 2 
‘Rug 19) 2-2 ee + pept7-22 2 ee — 
: (Acre 929) one + | ag ‘Mik Biscoreteeee + | 7 
9e A Sept. (8 ewe 4 0) Sepia. ee = 
Sentilles 2 2. eres sf (Sept. bis-2-. sees + 
Sept. 192 ee + 34_. [se 19/2 eee = 0 
20_- ees IED ee af | j | Sept./22) Se + 
j Airs ee ee + 35..|[BePt Bates Bee + 
21.. ee 21). eae + i | Sept 22). 220s aa ees _ 
Ange 2712-2 ese a Sept.10). 22... 2-28 + 
ae BD) aoe eee ar 9 || 36.- Pept cane eee + | 0 
Sept: 26 2.5... oss 3 Septieewccecsa cease + 
PVC ys aera te Sa a + \ SOUtIe osonno sees + 
23. lave. Os ee = 2 Sept de sacs he 4 
Sephii6:...c2 ae ae 87__ fen 1 | i as — 2 
PIR 2D (ene ae | Sept::20.4-52- ee _ 
Sept: 2 Sees ee tt ll ss eae |: eeeeaeees + | : 
#A.- {Sept. 7 __.--------- + | Ol); ml eptage es eee + iI 
Sentwiatel ero + | Sontb tsa! = 
Septs .-5 eee ci 39__ [sn ON. ard ne = 0 
HA TIBOIIG 2 Cee + } Septsso 2 ee + 
pet Aruar: BO acs —- | ; i 162 Sect ee + 
Sept 18-25 ss + ) 40--l) Sept. 19 ______._---- _ 2 
Sentqeacoe- cee = Sept22 ee ~ 
Aur 6. 2 + Septseliz- sas ee + 
26.- i De. Sa eee ant 0 al-- [sn De AR Sed ae Oat oe + 0 
Septail/s. 22s ar Sept0 eee see + 
Sentegy).. 2:2 + jo lee 1 ae Ee aaa, + ? 
(Arde 26 22 eeee + Och. 82 ee + 
Aue es, eater! of Sept 20 cee cn ree 7 4 
Sept.Si. 22-4 Soe Air 43__ Oct. RSs 36-- 22k _ “ 
27--'Sept. 9 ..-.-------- a 0 Oetr 2) -oe cee ee a 
Sept-20) 2282-2222 + Oct! Abi. 25 = 
Sept.7 ...2.2525-50 = re oe BO lope coco SF 0 
Sept. 26,-4scceeees + | Ochiit = + | 


IX, B, 1 Willets: Treatment of Entamebiasis 115 


TABLE V.—Cases treated with neosalvarsan. 


(Dose 0.9 gram.) 


| 
‘ Laboratory exami-; Treat- | Number | p Laboratory exami-| Treat- | ree 
| nations. ment of con- | nations. ment. of con- 
| on Te Goel aaa ee 
i | 
| Blasof laboratory Neue laboratory 
Date. Result. Date. nations Date. Result. Date. nations 
i before dis- before dis- 
charge. | charge. 
; al if |=: aan (is ee | iy Op Seger |S Sad 
1913. 1918. | | 
1 (es 4 =f Jan. 5 -- 1 Aug. 26 ta 
—— ll Jan. 9 _ Jan. 9 _- Bas Sept. 5 — Sept. of : 
x bere 3 + \mtay 3 4 Sept. 10 = Sept. 11_ 
— i May 5 ni ept. 24 _ 
Maps isnt Eee ct Sept. 1 oe 
| Aap) 40) eb =F Sept. 13 = 
7 ¢ Aug. 22 = Sept. 15 = 
Aug.27| — || Aug.9_- Sorel lentigo mes 7 fee htc B 
aoe Sept. 1 = Aug. 24. fi Sept. 17 es 
Sept. 5 = Sept. 11-_ Sept. 23 = 
Sept. 10 — Sept. 3), + 
Sept. 17 = Sept. 18 = 
Sept. 23 = (eos Sept. 24 aad PONE é 
Aug. 23 “b Sept. 26 | = 
‘ Sept. 6 an Sept. 1 _/] > Sept. 3. + 
Cn Sept. 10 = Sept. 8 _|| Sept. 6; + eae 
Sept. 12 | — {| 8.___}Sept. 16 | + Soe > 2 
| Sept. 23 | _ F 
| SS Ce Sept. 29 | _— 


TABLE V1.—Comparison of results of treatment in dysenteric cases. 


| 
Treated with | Treated with 


ipecac. | emetine. 
Cases. aie ane 
Number. | Per oe See Per cent. 
: esti ier ets Seco 
POTEET a ot A ls RG a ee [peat Ss | 1.000| 9.1 
Recovered symptomatically ......_...-.-..--------------- 10.00 | 62.5 | 10.000| 91.9 
Improved symptomatically___._____._-._-__----_----__--- 4.00 | PASS Sp sa gad | 
Unimproved symptomatically ___.______________________-- 2.00 | HONG eet ae. Se 
Discharged: | 
Negative microscopically______.____.______._______--- 


Positive microscopically____.-_.-.-_.__ ...-_-.-------- 
With two consecutive negatives® ___________________- 
Average days for symptomatic recovery after treat- | 
ment began. 
Average days to first of two final negatives _____________ ) 
Average grams of drug required for symptomatic re- 
covery. 
Average grams of drug to first of two final negatives ___ 


* By actual examination and by estimation, see Table VIII. 


116 The Philippine Journal of Science 1914 


TABLE VII.—Comparison of results of treatment in dysenteric cases with 
those obtained by Rogers (5). 


| } Died of — Discharged symptomatically— 
Dysentery. Not cured. Cured. 
B ra | 
Drug and authority. | After Sauene ppther 
iseases. 
Within), sae Num- Num- 
Sidace! Aue} new Per cent. Fan Per cent. 
hex! Per cent. 
SA an | ae eee 
Ipecac: | 
Rogens) -23--425- > b= 4 % 2639) Seen s Se 6 23.1 13 50. 0 
Ourselves... - 288 95 Se eee [oa ee ee ee 6 37.5 10 62.5 
Emetine: 
21 100.0 
10 91.9 


. 8 Exclusive of those who died within three days and of diseases other than dysentery. 


Average number of days from beginning of treatment to symptomatic 
recovery: 


Ipecac. Emetine. 
Rogers 11.4 2.35 
Ourselves 8.1 3.60 
Average grams of drug given during this period: 
Ipecac. Emetine. 
Rogers 27.1 0.130 
Ourselves "16.3 0.287 


8 Exclusive of amount given to 2 children. 


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abt tind 3. 


NERVE DEGENERATION IN FOWLS FED ON UNHUSKED RICE 
(PALAY) 


By R. B. GIBSON and ISABELO CONCEPCION 


(From the Laboratory of Physiology, College of Medicine and Surgery, 
University of the Philippines) 


One plate 


The observations of Fletcher’ and of Fraser and Stanton? 
have shown that diets consisting chiefly of polished rice are the 
common cause of beriberi in the Orient. If the white rice, 
however, be replaced by the unmilled variety, the disease does 
not develop. Substitution of rough rice for the polished article 
and additions to the dietaries of the public institutions of the 
Philippine Islands and of the Philippine Scouts are stated to 
have eliminated beriberi from these organizations.® 

It is generally believed, then, that among rice-eating Orientals, 
beriberi is due to the use of rice which has been deprived of 
its cortex. 

Further confirmation of this idea is obtained by the discovery 
that rice polishings or extracts of these, when fed with the 
milled grain, will protect against polyneuritis in fowls. The 
isolation of certain “vitamines” from rice polishings and yeast 
are reported in the studies by Funk,® by Suzuki, Shimamura, and 
Odaki,® by Edie, Evans, Moore, Simpson, and Webster,’ and by 
Vedder and Williams.‘ These substances, extracted in relatively 
minute amounts from large quantities of material, have a prompt 
curative effect in fowls with polyneuritis. This evidence in- 
dicates that beriberi is due to the lack of the polished rice in 
accessory substances which play a role of extreme importance in 
normal nutrition. 


*Lancet (1907), 1, 1776. 

*Lancet (1909), 1, 451; Studies from Institute for Medical Research. 
Federated Malay States (1909), No. 10. 

* Heiser, This Journal, Sec. B (1911), 6, 229; Chamberlain, ibid. (1911), 
6, 138. 

“Schaumann, Beth. z. Arch. f. Schiffs- und Trop.-Hyg. (1910), 14, 325; 
Fraser and Stanton, Studies from Institute for Medical Research. Feder- 
ated Malay States (1911), No. 12; Chamberlain and Vedder, This Journal, 
Sec. B (1911), 6, 251; Chamberlain, Vedder, and Williams, ibid. (1912), 
Tip BEE : 

* Journ. Physiol. (1918), 46, 173, and earlier papers. 
* Biochem. Zeitschr. (1912), 43, 89. 
" Biochem. Journ. (1912), 6, 234. 


"This Journal, Sec. B (1918), 8, 175. 
119 


120 The Philippine Journal of Science 1914 


There is, however, evidence to show that the use of unpolished 
rice will not absolutely prevent beriberi in man. Thus Strong 


and Crowell,® in reporting their experiments on Bilibid prisoners, 
say: 


In Group III, where the diet consisted largely of red rice, only 1 (No. 13) 
of the 6 developed rather marked symptoms of beriberi, while 1 (No. 18) 
developed only slight cardiac symptoms. In Nos. 14, 15, 16, and 17 no 
symptoms at all of the disease developed. In No. 13 the most striking 
symptoms suggestive of beriberi were pain and tenderness in the epigas- 
trium, symptoms suggesting paresthesia, epigastric pulsation, cardiac dis- 
turbances and dyspnea, and marked diminution and almost disappearance 
of the knee jerks, so that it was very difficult or impossible at times to elicit 
them. The condition of this individual, at the time that his diet was 
changed, certainly led one to believe that had the diet been persisted with, 
a well-marked case of beriberi would have developed. 


Again Shibayama °° states: 


Experiments * * * were carried out in coal mines where the miners 
had been yearly affected most severely and in a fishing village where the 
inhabitants had also suffered severely. In both places the inhabitants were 
divided into groups consisting of a certain number of persons (usually 100) ; 
during the beriberi season (seven months from the beginning of April to 
the end of October) one group was provided with cured rice,” another with 
the mixed diet consisting of rice and barley, while a third was given white 
rice as a control, the object being to determine which group provided the 
largest number of patients. * * * The experiment was carried out 
twice in each place, and showed that neither the cured rice nor the mixed 
diet of rice and barley is able absolutely to prevent the disease, though they 
seem to play some part. 


While considerable success has obtained in the treatment of 
infantile beriberi with rice polishings or with extracts of the 
bran,’* cases in adult patients are apparently more resistant. 
Vedder and Williams * report 3 adult cases, and they believe 
that the administration of their preparation of extract of the 
rice polishings— 


is capable of dissipating the dropsy in cases of wet beriberi and of promptly 
relieving the attacks of cardiac insufficiency, but that this extract is in- 
capable of curing the paralysis in cases of so-called dry beriberi. 


* This Journal, Sec. B (1912), 7, 404. 

* Journ. Trop. Med. & Hyg. (1918), 16, 284. 

* “Before the exact origin and method of preparing rice capable of caus- 
ing beriberi was known, it was termed ‘uncured,’ while that which did not 
produce the disease was called ‘cured’ rice.” [Aron, This Journal, Sec. B 
(1910), 5, 82.] Evidently red or unpolished rice is meant in this case. 
[Footnote 11 is ours.—R. B. G.] 

“Gabriel, Revista Filipina (1911), 2, 441; Chamberlain and Vedder, 
Bull. Manila Med. Soc. (1912), 4, 26. 

* This Journal, Sec. B (1918), 8, 175. 


— 7 


eee ee _— 


1X, B,1 Gibson and Concepcion: Nerve Degeneration 121 


They believe that Funk’s base, administered in 2 doses each 
prepared from 10 kilograms of rice polishings, “greatly relieved, 
if not cured’ the paralysis in one case. These investigators 
believe that the large amount of material necessary to yield any 
appreciable amount of the protective substances is due to the 
fact that there is considerable loss in the chemical manipulation 
in the course of isolation. We understand that the treatment of 
beriberi cases in the Philippine General Hospital with rice polish- 
ings or preparations of the rice bran is not followed by the 
uniformly satisfactory results which should be expected from the 
theoretical view of the etiology of beriberi. 

From the evidence presented, then, there are cases in which 
beriberi apparently has developed, even though unpolished rice 
is the chief constituent of the dietary. Furthermore, the 
therapeutic use of the rice bran or extracts and preparations 
of this has not given the specific results for man which we have 
expected from theoretical considerations of the etiology of the 
disease, although the mortality has been greatly reduced.’* It 
seems to us that the facts can only be interpreted in one way. 
The unpolished rice, per se, affords only partial protection against 
beriberi. In other words, the rough rice does not contain in 
the cortex the protective substance in sufficient amounts to 
prevent the development of beriberi in the more susceptible 
individuals. Experimental evidence for this conclusion is given 
in the following experiment. 

Six fowls were fed on palay, 3 for two months, 2 for three 
months, and 1 for four months. They developed no symptoms 
of neuritis. The sciatic nerves, when stained by the Marchi 
method, however, show distinct degenerative changes in every 
case. The results are summed up in Table I. 


TABLE I.—Fowls fed on palay. 


| No. Period. Degeneration. 
Months 
1 (6 2| + + + Fairly pronounced. 
2 (82) 2 | + Mild type; distinct lineation of myelin with few degen- 
erated areas. 
3 (83) 2 | + + Distinct degeneration. 
4 (80) 3 | + -+ + + Very pronounced degeneration. 
5 (34) 8/| ++ + + Very pronounced degeneration. 
6 (29) 4 


+ + Distinct degeneration. 


“ Braddon, Journ. Trop. Med. & Hyg. (1918), 18, 282. 


122 The Philippine Journal of Science 1914 


Clark * has shown that degeneration may be noticed within 
seven days on a polished rice diet in fowls. Such degeneration 
is, however, only very slight. The pictures of the sciatics in our 
palay-fed fowls correspond to that obtained from those fed for 
two weeks or more on milled rice. The degeneration in fowls 
4 and 5, in fact, is more pronounced than can be observed in 
some fowls which have died of rice polyneuritis. 

It would seem, then, that the substitution of rough rice for 
the polished article cannot be completely protective against beri- 
beri in all individuals. We do not mean to say that this sub- 
stitution has not practically eliminated beriberi where it has been 
undertaken or that the use of rice polishings is not without 
great therapeutic value. But in view of the accumulated evi- 
dence, we can say that the addition of other foodstuffs to a diet 
of unpolished rice is essential to meet the normal nutritive re- 
quirements of the body. 


* Vedder and Clark, This Journal, Sec. B (1912), 7, 423. 


ILLUSTRATIONS 

PuaTe I 
_ Fibers from the sciatic nerve of fowl 30 fed on palay for three 
months. 
2. Fibers from the sciatic nerve of fowl 31 fed on palay for two 
‘months. 


123 


He Nea nat. te. Rw sie ea 
pean vei Heke tii3 ‘4 ae 


Taos 


SAMY  E ert Ai 
hares 


we} yale er Net: Ge twee’ We ated, oipdiut o>. ret By 


“alee of Ge) 77.1 i oyruce middie aalt Iteod evedit 


GIBSON AND CoNncrpcioON: NeRvVE DEGENERATION. | [Pum. Journ. Scr., IX, B, No. 1. 


Fig. 1. Fibers from the sciatic nerve of fowl 30 fed on palay for three months. 


Fig. 2. Fibers from the sciatic nerve of fowl 31 fed on palay for two months. 


PLATE I. 


SIMULTANEOUS METHOD OF INOCULATING CATTLE AND 
CARABAOS WITH SERUM FROM ANIMALS THAT 
HAVE BEEN RECENTLY IMMUNIZED 


By ARCHIBALD R. WARD and FREDERICK WILLAN Woop 


(From the Veterinary Division, Bureau of Agriculture, Manila, P. I.) 
Three plates 


The earlier literature concerning antirinderpest serum con- 
tains frequent references to the idea of increasing the protective 
power of the serum by successive injections of virulent blood 
into the animal producing the serum. The idea is emphasized 
that the serum of such hyperimmunized animals is preferable to 
the serum from animals that have merely been immunized nat- 
urally or artificially. 

Braddon(1) states that in South Africa the method of injec- 
ting uninfected animals with defibrinated blood of animals which 
had recently recovered from pest, as a preventive method, was 
abandoned in favor of Turner’s method of using serum from 
animals that had received successive and increasing doses of 
virus. 

Turner (2) states that in the Transvaal and Natal an immune 
animal was injected with 100 cubic centimeters of virulent blood, 
and after all reaction had ceased it was bled. This blood, when 
defibrinated, was injected into a susceptible animal which was 
smeared on the muzzle with virulent material and placed with 
animals suffering with rinderpest. This method of simultaneous 
inoculation, using blood prepared at the time, seems to have been 
abandoned in favor of one employing a serum requiring more 
elaborate preparation. 

Since the use of serum from hyperimmunized animals came 
into general use in simultaneous inoculation, Gibson(3) is the 
first writer known to us, who has questioned the necessity of 
hyperimmunizing serum-producing animals. 

Shealy (4) makes a similar observation to the effect that the 
results obtained with the serum prepared from animals after 
recovery from an attack were found to be just as good as when 
the animal was not bled until it had been hyperimmunized. ~ 

Subsequent to the completion of the field work described 

125 


126 The Philippine Journal of Science 1914 


herein, there has come to notice the work of Holmes(5) which 
gives additional information regarding the overestimation of 
value of serum from hyperimmunized animals. He concludes 
that the serum obtained after natural recovery or after an im- 
munizing reaction is little inferior in potency to that taken after 
the process of hyperimmunization. 

Holmes further points out that satisfactory virulent blood 
may be obtained from an animal suffering a modified attack due 
to the injection of serum. Thus, the animals producing virulent 
blood recover, where formerly they were sacrificed. 

The work of the present writers was undertaken with a view 
of reducing the cost of serum to a point that would warrant more 
extensive use of simultaneous inoculation. The serum formerly 
produced by the Bureau of Agriculture cost, delivered in the 
field, 24 pesos’ per liter. That this is not excessive is shown by 
the fact that in the Transvaal the cost was 25 pesos per liter. 
Small quantities purchased by the Bureau of Agriculture from 
the Pasteur Institute at Nha-Trang, French Indo-China and from 
the Experiment Station for Animal Diseases, Tokyo, Japan, 
were charged for at the rate of 47.89 pesos and 34.40 pesos per 
liter, respectively. With serum costing 24 pesos per liter and 
with the doses employed by us, the expense in simultaneous 
inoculation for the one item of serum would slightly exceed 8 
pesos per animal. 

In deciding to try the use of serum from animals that had 
just been immunized instead of employing expensive serum from 
the so-called hyperimmunized animals, we were guided by 
several considerations. One of us, when superintendent of the 
serum laboratory of the Bureau of Agriculture, carrying on the 
routine work there of immunizing susceptible animals for serum 
production, employed serum in several cases from animals that 
had recently reacted. The results were as satisfactory as if 
hyperimmune serum had been employed. Further, previous ex- 
perience(6) had shown that the severity of the immunizing reac- 
tion could be controlled by the amount of serum employed. 
Thus, if the serum drawn in the field proved to be appreciably 
low in protective power, the fault could readily be corrected. 

The work was inaugurated in Ilocos Norte Province on 
carabaos belonging to the Calamba Sugar Estate, and intended 
for shipment to their estate at Calamba, Laguna. When the 
immunization was begun, there was available a supply of serum 
from hyperimmunized animals only sufficient for the first three 


* One peso Philippine currency equals 50 cents United States currency. 


x 


1x,B,1 Ward and Wood: Inoculating Cattle and Carabaos 127 


lots of carabaos as indicated in Table I. All animals immunized 
subsequently to these received serum drawn in the field. 

The animals while undergoing immunization were confined in 
large sheds erected by the owners of the carabaos, who likewise 
fed and cared for the animals during the twenty-five days 
required for immunizing. 

Each animal to be immunized, while being injected, was con- 
fined in stocks made of bamboo. It received a hypodermic in- 
jection of the dose of antirinderpest serum which had been 
decided upon as sufficient. This standard dose was administered 
_ uniformly without reference to the size of the animals. 

In the case of the carabaos belonging to lots 1, 2, 3, and 4 shown 
in Table I on page 127, it will be noted that gradually increasing 
doses were employed until there were obtained results thoroughly 
satisfactory in regard to death rate and severity of reaction. In 
determining the dose, it was considered advisable to use an 
amount of serum necessary for the most susceptible animal likely 
to be encountered, for there exist no means for predicting 
susceptibility. After the immunization work was under way, 
the virulent blood was invariably obtained from an animal 
undergoing immunization at a stage corresponding to the third 
day of febrile temperature. Thus, there was no expense for 
susceptible cattle for maintaining a strain of virulent blood, and 
no trouble of finding natural cases of rinderpest in the vicinity 
was necessitated. 

In all cases, before virulent blood was employed, it was 
examined microscopically for the presence of the trypanosomes 
of surra in fresh and stained preparations. 

One hundred ninety-six of the 429 animals in Table I, 
which showed no febrile reaction, were reinjected with 10 cubic 
centimeters of citrated virulent blood. This was done in case 
that the original virulent blood had been inactive when employed 
or some accident had prevented its introduction. Up to date, 
only three distinct reactions from such injection have been 
observed. 

In the preparation of serum in the field, between 2 and 3 
liters of blood were drawn from the jugular vein of all recovered 
animals without reference to whether or not they had reacted 
to the virulent blood. In a previous(6) paper it has been shown 
that serum from reactors is somewhat more potent, but with 
the dose employed this feature was not of sufficient value to 
warrant exempting nonreactors from the bleeding. 

No blood was drawn from very old or very young, nor from 
pregnant animals. These exceptions reduced the number bled 


128 The Philippine Journal of Science 1914 


to 72 per cent of the animals immunized. An average of 2.8 
liters of blood per animal was drawn from 305 animals. 

The technique employed in drawing blood for serum produc- 
tion varied in no essential particular from general practice. 
An autoclave heated by a gasoline torch was employed for 
sterilizing the instruments and bleeding flasks. No abscess 
formation has followed the bleeding. Large hematomata form 
occasionally, but are soon resorbed. 

Blood was allowed to stand in the bleeding flasks immersed in 
running water in the bed of a shallow stream for from twenty- 
four to thirty-six hours, after which the serum was decanted 
into a large graduated vessel and 0.5 per cent of carbolic acid 
or formalin was added as preservative. The serum was then 
stored in 15-liter demijohns tightly corked. Serum prepared 
in this way has kept well for several weeks. The only trouble 
encountered was due to the fact that serum sometimes assumed 
a gelatinous form upon standing. 

The serum thus prepared was ready for use, there being omit- 
ted the expensive operations of centrifugalization, filtration, and 
rebottling. There is nothing connected with the preparation 
of serum under field conditions which requires an expensive per- 
manent plant. Experience has shown that the essential asepsis 
can be readily attained in a temporary structure made of grass 
and bamboo. 

It should be noted that no ice was available for refrigeration. 
The necessity of holding serum for a long period was obviated 
by preparing it with reference to the time that it would be 
needed. 

The results obtained during the immunization of the carabaos 
at Laoag are shown in Table I. In compiling the data regarding 
reactions, there has been adopted as a standard of minimum rise 
of temperature to be counted as a reaction a rise to 39° C. on 
two successive days, occurring between three and twelve days 
after inoculation. This standard is wholly arbitrary and some- 
what unsatisfactory when applied to cases closely approaching 
it; however, none more generally useful is known. In the case 
of the animals in question, the dose employed was such and the 
average resistance of the animals of such a degree that elevation 
of temperature does not figure prominently. Other symptoms 
of rinderpest such as diarrhcea and inflammation of the conjunc- 
tive occurred, but this occurrence was not uniformly recorded on 
the temperature charts. 


a 


x,B,1 Ward and Wood: Inoculating Cattle and Carabaos 129 


TABLE I.—Record of immunization at Laoag of carabaos belonging to the 
Calamba Sugar Estate. 


Febrile reac- Loss after 
Lot N. Number | Date of | Serum Pea, oh, Number Fs OCR 5 
at | injected. | injection. | per head. P released. P 
} No. Ge No. Oe 
cent. cent. 
C.c. | ! 
ge ORL NAR tS 8| July 28 200 4} 50.0 6 2| 25.0 
| Se ad Se | 70| Aug. 11 300! 28) 40.0 70 | 0] oO 
ie ae se tee BN aye Ne | 63 | Aug. 23 300 | 48| 76.0 60 81 47 
fi. Sys a Soe Niel OE 65 | Aug. 28 350 | 43 | 66.0 65 0] oO 
GAs Roe te See) ee 27! Sept. 8 350 | 15| 56.0 27 | 0} oO 
Ga eeaireeire ul ad 2 15 | Sept. 12 | 350; 13] 86.0 15 | 0] 0 
Tass 10 | Sept. 15 350 | 20.0 10 | 0| o 
i 26 | Sept. 24 | 350 | 4] 16.0} 26 | 0} 0 
ti ee oe 31 | Sept. 26 | 350 / 6| 19.0 31 ON'0 
Te «TS 51| Oct. 3 350 22| 43.0 51 0}; 0 
ill, coo 63 | Oct. 9 350 42| 66.0 62 1] 15 | 
Tataleee es > dog i teae ss eee” Ee ealeeti | 227| 52.9 423 6| 1.4 | 


It will be observed that all of the deaths, except one, occurred 
in the first three lots. In the case of those animals only, serum 
made in the laboratory from hyperimmunized animals was used. 
Increase of the dose of serum from 200 cubic centimeters to 300 
cubic centimeters practically stopped losses. 

It is believed on the basis of the previous work(6) and that of 
Holmes(5) that an animal may be immunized by simultaneous 
inoculation without showing either fever or symptoms. There- 
fore, it is thought possible that more susceptible animals may 
have been immunized than shown in the table. 

Table II gives data concerning the number of animals in each 
lot which attained maximum temperatures of a degree corre- 
sponding to four classes chosen arbitrarily. 


TABLE II.—Febrile reactions among carabaos at Laoag. 


| iMaSharnn, eae tals ET aiiiss vi) Totals for each 
Aelconsol ae pele / 7!/sio9lwlua temperature class. 
} | Number. | Per cent. 
PUOMUVEN eo ee oie ae 4| 15 (| 45 | 33 | 11} 10 Dp 2 2b |-20 158 269.6 
PACS a a | oj 12] 1] 8| 3] 2] 1] 1] 2] 6] 44 50| 422.0 
BP G99. Ae ee eta ol Baa) teal oy e] at il s 19! 98.3 
“5G Chet ae ee a ae | 4142115! 22) 12! 2 8! 22 | 25 | 29) 21 202 b47.1 
Nonreactors -.=-.--.-=..---. per ee | eed se eel eee Reet ES ge SE Ne Ie a ae | 
Total number of ani- | sere | 
yo | Saal ee eae een | 8| 70 | 68| 65 | 27 | 15 | 10} 26 | 31 | 51 | 63 CL i eee 


* Per cent relation to 227, the total number of reactors. 
> Per cent relation to 429, the total number injected. 


It will be noted that 46.3 per cent of all failed to reach 39° C. 
and are not reckoned as having shown a febrile reaction in 
accordance with the standard described above. Comparatively 
few attained a temperature between 39° and 39°.5. 

124293——9 


130 The Philippine Journal of Science 1914 


In considering the results of immunization, there arises a 
question regarding the general susceptibility to rinderpest of the 
animals in the district where the work was carried on. In as 
much as a very strict system of quarantine and inspection of ani- 
mals was in force in the same province at practically the same 
time, some data can be presented. During the period from Sep- 
tember 20 to December 27, 1913, in the same province and in 
almost adjoining municipalities, 94 cases of rinderpest were dis- 
covered of which 36 died, a number corresponding to 38.3 per cent. 
This in our experience is a moderately low death rate; we have 
observed it to vary from 33 per cent to 100 per cent elsewhere. A 
low death rate is generally considered to indicate a high resistance 
of the animals to the disease or a low virulence of the strain of 
virus harbored by the animals in thecommunity. Therefore, with 
regard to the death rate prevailing naturally at the time, the work 
of immunization was conducted under favorable circumstances. 

Subsequent to the work of immunization at Laoag, extensive 
work of the same kind for the general public has been carried 
on at Dingras and at Solsona by Dr. J. R. Burns. 

From October 11, 1913, to January 25, 1914, 1,657 animals 
were immunized in these municipalities, not counting 954 ani- 
mals injected previous to this date but not completed. More 
would have been injected during the period but for the fact 
that 65 liters of serum have been shipped to Manila for use in 
starting the work elsewhere. The total number of deaths 
during immunization for the period was 23, which corresponds 
to 1.3 per cent of the number injected. 

As the work has progressed, the expense has been materially 
lessened. The immunization of 429 animals at Laoag cost the 
Bureau 3,994.11 pesos, about 9 pesos per animal. The work 
was essentially experimental in character and was conducted 
slowly and cautiously. 

Conditions of routine field work are more closely represented 
by the work in Dingras and Solsona. For the period October 
11 to December 31, 1913, the following expenditures were made 


by the Bureau: 
October. 
Salary of 1 veterinarian, 5 days, at 333.33 pesos per Pesos. 
month 55.55 
November and December. 


Salary of 1 veterinarian, at 333.33 pesos per month 666.67 


Wages of 10 laborers, at 20 pesos per month 400.00 
Wages of 2 laborers, at 45 pesos per month 180.00 
Expendable supplies furnished during period 123.55 


Total 1,425.77 


x,B,1 Ward and Wood: Inoculating Cattle and Carabaos 131 


No item is included to cover cost of autoclave, bleeding table, 
and like apparatus which are permanent in character. Auto- 
claves complete with gasoline heating apparatus are obtainable 
for 300 pesos and bleeding tables cost 93 pesos each. At all 
times during the work, several veterinarians and inspectors 
have participated in the work to receive instruction in technique, 
but as their presence was unnecessary for doing that work 
their salaries and traveling expenses have not been included 
in the statement of expenses. Only five days of the time of one 
veterinarian during October is charged for, as he was employed 
on the work in Laoag for the remainder of October. 

Estimates of the number of animals immunized per month 
and the cost based upon the early period of the work do not 
constitute an index of progress under conditions when it is 
thoroughly under way. A scarcity of serum during the early 
stages limits the economical employment of labor. Thus from 
October 11, 1913, to January 25, 1914, an average of 473 animals 
per month was immunized at a cost of approximately 1.50 pesos 
per head. During the four weeks beginning December 28, 790 
animals were released. However, during this same month, 1,056 
were injected, which will be approximately the number of im- 
munized animals that will be released a month hence. Esti- 
mating the expenses for salaries, labor, and supplies at 685 pesos 
per month, which is the average for November and December, 
it is concluded that the immunization of the 1,056 animals in- 
jected during the month beginning December 28 cost the Bureau 
65 centavos apiece. It is believed that this rate of progress 
can be improved by instituting certain changes affecting the 
amount of travel. 

Had the serum employed in Dingras and Solsona been pro- 
duced in a serum laboratory, there would have been an added 
expense up to January 25 of 21,932 pesos, reckoning the cost at 
24 pesos per liter. This would have brought the expense up to 
8.40 pesos per animal, a cost deemed prohibitive. 

At the inauguration of the work, the owners of the animals 
to be immunized constructed the group of laboratory sheds 
shown in Plate I, fig. 1. The bleeding is done in the partially 
inclosed shed in the right background. Plate I, fig. 2, shows a 
carabao in this shed, restrained for bleeding. The first stage 
of placing an animal on the table is shown in Plate II, fig. 1, and 
the method of bleeding in Plate II, fig. 2. Both of the last two 
operations were photographed in the open air in order to obtain 
better light. The veterinarian holding the needle and one assist- 


182 - The Philippine Journal of Science 1914 


ant customarily stand on the opposite side of the animal. Other 
parts of the group of buildings in Plate I, fig. 1, house the office 
and general workroom. The bottles of blood after drawing and 
while the serum is exuding from the clot are placed in the 
stream bed in the inclosed building in the foreground. Adjoin- 
ing these sheds are others for accommodating animals during 
immunization, the construction being of the cheapest character 
and primarily designed merely to afford complete shade. An- 
other shed in the group shown in Plate III, fig. 1, contains stocks 
for restraining the animals during injection. The process of 
injection in this shed is shown in Plate III, fig. 2. 

Serum is all prepared at the central laboratory and all im- 
munized animals are brought there for bleeding. Immunization 
is carried on in other localities, some of them 10 kilometers 
distant, serum from the laboratory being sent out in demijohns. 
In every case, the owners of the animals have cheerfully con- 
structed the necessary sheds. When these are needed, each 
man brings with his carabao one or two bamboo poles and a 
bundle of grass, and assists in the construction. 

Each animal immunized is branded on the right shoulder 
with numbers designating the province and municipality. All 
animals immunized in Ilocos Norte bear the number 1 and to 
the side of this is placed a number referring to the municipality. 


Thus the brand 11 indicates Laoag; 12, Dingras; and 13, Sol- 


sona; all in Ilocos Norte Province. Further, the animals in 
each municipality bear serial numbers which are entered on 
the immunizing records, with owners’ names. 

A certificate of immunization is issued for each animal when 
released. This contains a printed outline of an animal, upon 
which are indicated brands and other distinguishing marks, 
together with description, name of owner, etc. 

Animals accompanied by a certificate of immunization are 
exempted from any quarantine for rinderpest that is enforced 
by authority of the Director of Agriculture. 


CONCLUSIONS 


1. The writers believe that their experience with simultaneous 
inoculation with blood drawn in the field has demonstrated that 
there is no necessity for maintaining an expensive permanent 
laboratory and herd for the production of antirinderpest serum 
from hyperimmunized animals. 

2. A radical reduction in the cost of serum production has 


ee ee 


1x,B,1 Ward and Wood: Inoculating Cattle and Carabaos 1383 


been effected, and in consequence the possibility of extensive 
employment of simultaneous inoculation in combating rinder- 
pest has been demonstrated. 


REFERENCES 


(1) Brappon, W. LEoNARD. Report to the Government of Negri Senibilan 
on an experimental investigation into the methods of protection of 
buffaloes and cattle against rinderpest. Kuala Lumpur (1902). 

(2) TuRNER, G. Rinderpest in South Africa. Journ. Trop. Vet. Sci. 
(1906), 1, 269. 

(3) Gipson, ADAM. A method of dealing with rinderpest in the field. 
Journ. Trop. Vet. Sci. (1910), 5, 28. 

(4) SHEALY, A. S. Some facts and ideas about rinderpest and their rela- 
tion to the Philippines. Proc. Am. Vet. Med. Assoc. (1910), 397. 

(5) Hoitmes, J. D. E. Rinderpest. Further investigations on questions 
connected with the economical production of antiserum. Memo. Dep. 
Agr. India. Vet. Series (1913), II, 2. 

(6) Warp, A. R., and Woop, F. W. Experiments on the efficiency of anti- 
rinderpest serum. Bull. P. J. Bur. Agr. (1912), 19. 


ya 


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aint PMY apie mt Bice itor? teint 


. trek 1g ti , £9e) by it Mise Ten Ee (Tt fos tow huecaeseeyn fC A 
a eshneeideg werent rian: lewis amy 


rere 
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wey eel 
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POR. cpg bpenel * a nGh of. Ad 
— sabett Pi ibe, nade BIT: . 
aia tp Rabie Lh aia ot tite bs + SiPAD 
TE C8 AY hin netintk Mae 
‘$000 00 Trt a Rae b 
vt aves we treprenenieng 


ILLUSTRATIONS 


PLATE I 


1. Field laboratory for the production of antirinderpest serum, Dingras, 
~ Tlocos Norte. 
2. Method of restraining animal for bleeding. 


PLATE II 


ts Method of restraining animal on bleeding table. 
. Operator drawing blood for the production of serum. 


PLATE III 


1. Shed in which animals are injected with virulent blood and serum. 
Operator injecting carabaos with virulent blood and serum. 


135 


wast 


ide eothegle ses hie upintiontegs 
1a vee. Le oh dekh add 582 fags: btn 


fui atest 


hetiraty a i hele tn Pi 
weol® tustaxty tlw somlbeniey 


WARD AND Woop: INOCULATING CATTLE AND CARABAOS.] [PHIL. JouRN. Sct., Vou. IX, B, No. 1. 


Fig. 1. Field laboratory for the production of antirinderpest serum, Dingras, Ilocos Norte. 


Fig. 2. Method of restraining animal for bleeding. 


PLATE I. 


WarD AND Woop: INOCULATING CATTLE AND CARABAOS.] [PHIL. Journ. Sct., Vou. IX, B, No. 


Fig. 1. Method of restraining animal on bleeding table. 


Fig. 2. Operator drawing blood for the production of serum. 


PLATE Il. 


“ty 


WarD AND Woop: INOCULATING CATTLE AND CARABAOS.] [PHIL. JouRN. Sct., Vou. IX, B, No. 1. 


Fig. 1. Shed in which animals are injected with virulent blood and serum. 


Fig. 2. Operator injecting carabao with virulent blood and serum. 


PLATE Ill. 


‘A FLORA OF MANILA 
By Ever D. Merri, 
"| Order No. 419. Paver, 490 pages, $2.50, 


postpald. 
sf « Praoti i ‘ complete flora the cul- 
a pean nthe Pailiseives: Descrip- 
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sary of technical terms, etc. 
evs mE | gocowur. ‘PALM IN THE PHI. 
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Soicles Inc 37. Paper, 149 pages, 30 plates, 
$1, postpaid. 


. the Water Relations of the Coconut 
et wees hucifera), The Coconut and its 


to’ te vib, The The ping. a 
As open 6 
idity, and The Principat, Inseots Attaok= 
‘ing the ‘onut, Palm. 


- INDO-MALAYAN Woops 
By Frep W. Foxwortay 


“NG No. 411, Paper; 182 pages, 9 
x ae plates, $0.50, postpa stpald. 
Indo-Malayan Woods, Doctor Fox- 
‘has brou ¢ together @ la 6 amount 
Fepeeret information concerning trees 
ing woods of economic value. . 


\ Pt 7 ., ie 
pre hs oA 
FRA oe istribution of each species 
aes eam D tyes Sb a descriptions are 


a 


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Sat a Ode ree PF 


ty reprint contains the following arti-. 


This is “the only recent abe to enu-) 
yr eniat of the Philippine .— 


PUBLICATIONS FOR SALE BY THE BUREAU OF SCIENCE, 
MANILA, PHILIPPINE ISLANDS—Continued 


ZOOLOGY—Continued 
' A MANUAL OF PHILIPPINE BIEDS 
By Ricwarp C. MeGkecon 


Order No. 103. Paper, 2. parts, 769 
,. pages, $4, postpaid. 


A Manual. of Philippine Birds contains 
in. compact form descriptions ‘of ail the 
known species of Philippine birds. The 
usual keys and diagnoses of orders, families, 
and genera help ‘the novice in identification, 


A CHECK-LIST OF PHILIPPINE 
FISHES ‘ 


By Davin Starr Jorpan and Rosert Bar. 
RICHARDSON 
Order No. 102. Paper, 78 pages, $0.75, 
postpaid. 


This list will be found a convenient guide 
to the synonymy of Philippine ichthyology. 
The nomenclature is thoroughly revised, and 
the distribution . of pet species within: the 
Philippine Islands is given. 


MEDICINE 


REPORT OF THE NATIONAL 
PLAGUE CONFERENCE 


Held at Mukden, April, 1911, under the 
" auspices of the Chinese Government, 


Edited by Extca Martini, G. F. Petam 
ARTHUR ars and RicHagp. P, 
TRONG 


483 pages, 18 plates (2 colored, 4 half- 
nes, 12 charts and maps) 


Order No. 416. Paper, $2.50; cloth, 
$3.50; postpaid. 


The proceedings of this International Con- 
ference and information gained therefrom, to- 
gether with the results of certain baote- 
riological investigations, constitute the pres- 
ent report. 

The Bureau. of Science of the Govern- 
ment of the Philippine Islands has been 

appointed sole agent for the distribution 
of the printed proceedings of the Interna- 
tional Plague Conference, 


&, waey ~ prices ARE IN UNITED STATES CURRENCY 


eae don these iplckenies may be sent to the BUSINESS MANAGER, 
Ws eutarnes JOURNAL OF SCIENCE, BUREAU OF SCIENCE, MANILA, P. L,, 
Or to a of the agents listed below. Please give order number. 


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_ Wm. Wesley & Son, 28 Essex Street, Strand, London, W. C., England. 

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; Aeowmiaancs. Sees & ae he es hapa me Calcutta, India. 


CONTENTS 


BARBER, M. A. Cockroaches and sata as Carriers of ‘the ‘Vibrios, et ciisls2) ae 
Asiatic Cholera _.. estas 
HEISER, VICTOR G. Reappearance of Plague in the Philippines after Bria earns 
_ an Absence of Six Years. Brief Description of the Outbreak, the 
. Methods Used to Combat it, amd the idaroncnntt Factors in its Tn- yin 8 
troduction ........ he RON NY AY Ee od cs Fe Ua Mr EMS Nr gh ah EScudsy MDA ale ahs 
BOYNTON, WILLIAM HUTCHINS. A Preliminary Report of Repent er 
ments on the Cultivation of the Virus of Rinderpest in Vitro...) | 
BOYNTON, WILLIAM HUTCHINS. An ri oceaeai Case of siiderpeet ti in 
@ Cara bao ie stk i ech a Si oe i ead OR ian cca ae ae is RS CON 
WARD, ARCHIBALD R., WOOD, FREDERICK WILLAN, and BOYNTON, ye 
WILLIAM HUTCHINS. Experiments upon the Transmission . of 
Bea ONT SSE ee ge alc ha ete are eo te a 
WILLETS, DAVID. G. Intestinal Parasitism, Particularly Entamebiasis, 
in Patients of the Philippine General Hospital, Manila, P, T_........ 
WILLETS, DAVID G. Preliminary Report on the Treatment of Enta- 
meébiasis with Ipecac, Emetine, and Neosalvarsan at hat ican ; 


Géneral; Hospital, Manila; Pos ae hs ae ea ; 
GIBSON, R. B., and CONCEPCION, ISABELO. . Nerve PT in’ 
Fowls Fed on Unhusked Rice (Palay) 2.03000 TaN 


WARD, ARCHIBALD R., and WOOD, FREDERICK WILLAN, suhariee° 
neous Method of Inoculating Cattle and Carabaos with Serum from | 
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THE PHILIPPINE ; 
~™Onal Wiusee> 


JOURNAL OF SCIENCE 


B. TROPICAL MEDICINE ke 
Vou. IX APRIL, 1914 No. 2 


SANITARY SURVEY OF THE SAN JOSE ESTATE AND ADJACENT 
PROPERTIES ON MINDORO ISLAND, PHILIPPINE 
ISLANDS, WITH SPECIAL REFERENCE TO 
THE EPIDEMIOLOGY OF MALARIA 


By A JOINT COMMISSION OF REPRESENTATIVES FROM THE 
COLLEGE OF MEDICINE AND SURGERY, UNIVERSITY OF THE PHILIPPINES; 
BUREAU OF SCIENCE; AND BUREAU OF HEALTH 


Three maps 
MEMBERS OF THE COMMISSION 


Dr. W. E. MUSGRAVE 
Dean of the College of Medicine and Surgery; Chief of Staff, Philippine 
General Hospital; Chairman of the Commission 


Dr. HE. L. WALKER 
Acting Chief, Biological Laboratory, Bureau of Science 


Dr. THOMAS W. JACKSON 
District Health Officer, Bureau of Health 


Mr. CHARLES S. BANKS 
Entomologist, Bureau of Science 


Dr. ANTONIO VAZQUEZ 
Instructor in Surgery, College of Medicine and Surgery 


Dr. P. GUTIERREZ 
Instructor in Medicine, College of Medicine and Surgery 


Mr. F. A. DALBURG 
Geologist and Topographer, Bureau of Science 


Dr. I. CONCEPCION 
Assistant, Bureau of Science 


Dr. SetH L. Cox 
Interne in Medicine, Philippine General Hospital 


Mr. ARISTON GUZMAN 


Assistant, Bureau of Science 
124316 137 


1388 The Philippine Journal of Science 1914 


CONTENTS 
. Introduction. 


. Geography and Geology. 
. The General and External Sanitary Problem. 
. The Internal Sanitary Problem. 
. The Mosquito Survey in and near San José. — 
. The Disease Index. 

(A) Laboratory examinations. 

(B) Clinical examinations. 
7. Special Reports. 

(A) A comparison of the spleen index with the microscopic examina- 
tion of the blood for malarial parasites in 1,003 persons. 

8. Summary and Conclusions. 
9. Recommendations. 
10. Appendix. 


aAnr wD ee 


1. INTRODUCTION 
By W. E. MUSGRAVE 


The property included in this survey lies on the west coast of 
Mindoro (fig. 1), and comprises from 210 to 260 square kilo- 
meters of territory. It is bounded on the one side by the ocean, 
on the opposite side by the mountains of the island, inhabited 
by the wild tribes of the country, while up and down the coast 
are located a number of very insanitary and unhealthy barrios. 
Along the ocean front, within banca distance of the property, 
are a number of small islands and inland points inhabited by 
thriftless, ignorant, and disease-infected settlers. 

For more years than can be remembered by the oldest living 
inhabitant or than may be traced in history, the place has been 
known as the “white man’s grave” and probably with a consider- 
able show of justification. During the last few hundred years, 
the Spaniards made several attempts to avail themselves of the 
fertile lands of this estate, and, in each instance, failed because 
of the unhealthful conditions and the high mortality among 
their employees. The Mangyans, wild tribes of the adjacent 
mountains, refuse to make more than transient visits to these 
lowlands, and have a superstition that, if they remain more 
than seven days without returning to the hills, they will become 
sick and die. So far as can be learned, malarial fever was then, 
as it is to-day, the principal cause of the high mortality rate 
among residents of this community, although other infectious 
diseases are about as prevalent as they are elsewhere in the 
Philippine Islands. 

To place this property, generally known as the San José Estate, 
in a satisfactory sanitary condition and subsequently to maintain 
it in a normal sanitary equilibrium will require an enormous 


— 


IX, B, 2 Sanitary Survey in Mindoro . 189 


amount of well-directed energy and a great deal of money, and 
when accomplished this work probably will be the greatest sani- 
tary achievement of a private corporation in history. 

At first glance, the problem of the Mindoro Company and the 
San José Estate would appear to be one impossible of accomplish- 
ment with the expenditure of a reasonable amount of money; 
and it is probable that, had the owners of the properties realized 


MINDORO ISLAND 


SCALE 


ae. a0 


Fic. 1. Mindoro Island, showing the location of San José Estate. 


the magnitude and the complexity of their sanitary problem in 
advance, the attempt to develop the agricultural resources of the 
country would have been delayed indefinitely. However, the devel- 
opments of the company already are of some three years’ dura- 
tion, enormous sums of money are invested, progress in sanitation 
has been made, and the present personnel of the directorate 
realizes that it is necessary to make its properties habitable 
in order to insure the success of the large agricultural under- 
taking. 


140 The Philippine Journal of Science 1914 


The sanitary problem of the Mindoro property compares 
favorably in magnitude with that of the Canal Zone. The total 
area of the Canal Zone is between three and four times as large 
as that of the Mindoro property, but the boundary lines and 
surroundings as well as the internal difficulties are so much 
greater in the Mindoro property than they are in the Canal Zone, 
that the sum total of the two problems should not be considered 
very different. In the Canal Zone, every interest is subservient 
to the digging of the canal. In the Mindoro property, drainage, 
irrigation, and other requisites for successful agricultural cultiva- 
tion must be maintained while the sanitary problem is being 
solved. In both places, labor must be imported to do the work. 
Panama is fortunate in being able to select a more healthful 
class of employees, because of an unlimited supply and consequent 
ability to enforce more rigid physical requirements. In Mindoro, 
the labor practically must come from other parts of the Philippine 
Islands, and, in consequence, is made up largely of Filipinos. 
The extreme difficulty in securing this class of labor in sufficient 
numbers has made it seem advisable for the corporation to dis- 
regard the health of applicants for work, and consequently un- 
healthy people—a large number of them even suffering from 
infectious diseases—have been, and constantly are being, im- 
ported into the estate. 

In Panama, due to the resources of a national government, 
it has been possible to protect the boundary lines both by sea and 
shore, and, with the abundant supply of labor and with the 
ability to enforce rigid physical requirements to applicants for 
positions, what may be termed the “external problem” of Panama 
is a very simple one compared with that of the Mindoro Company. 

If the enormous sums of money expended for sanitation in 
the Canal Zone are considered to have been economically admin- 
istered and necessary for the protection and safety in that area 
of property 16 kilometers wide and 64 kilometers long, it would 
not seem possible to solve the equally complex and difficult sani- 
tary problem of the Mindoro Estate within the bounds of any 
financial consideration consistent with the profitable investment 
of money. However, with the expenditure of funds inestimably 
small compared with that of Panama, very marked progress has 
been made in the sanitation of Mindoro, and it is believed that 
it is possible, under wise direction, to solve this problem with the 
expenditure of funds consistent with profit for the investors in 
this large agricultural undertaking. 

The properties of the Mindoro Company and the San José 
Estate are the most valuable on the west coast of Mindoro, and, 


IX, B, 2 Sanitary Survey in Mindoro 141 


by the reason of the attempted modern improvements, represent 
a large majority of the enterprises of the entire island. The 
majority of the population of this coast is included in the em- 
ployees of the above-mentioned companies, and the corporations’ 
sanitary problem, therefore, becomes the sanitary problem of 
the east coast of the island, a fact which must be recognized both 
by Government authorities and by the officials of the company if 
a successful sanitary administration of the affairs of these cor- 
porations is to be expected. 

The solution of the sanitary problem of the epidemiology 
of malaria in this peculiar environment, with labor selected from 
among oriental people from various infected centers, appeared 
to be so important that after due consideration and with the 
approval of the Honorable, the Secretary of the Interior, rep- 
resenting the Government, and the hearty and generous 
cooperation of Senator George H. Fairchild and other officials 
of the Mindoro Company and San José Estate a joint sanitary 
commission, selected from the personnel of the College of Medi- 
cine and Surgery, University of the Philippines; the Bureau of 
Science; and the Bureau of Health, undertook the sanitary and 
medical survey of this property. 

The list of the personnel of this commission has been given, 
and the findings and recommendations will be found in the sub- 
sequent pages of this report. 

After a brief preliminary consideration, the work of the com- 
mission was divided as follows: 


I. Topography and geology, by Mr. F. A. Dalburg. 
II. The external problem, a sanitary and medical survey, by Dr. W. E. 
Musgrave, chairman of the commission. 
III. The work of the internal problem was divided into four sections as 
follows: 

1. The internal sanitary survey, by Dr. T. W. Jackson of the 
Bureau of Health. 

2. Laboratory section, under the direction of Dr. E. L. Walker, of 
the Bureau of Science, assisted by Doctor CEST and Mr. 
Guzman of the same Bureau. 

3. Clinical section, under the direction of Doctors Vazquez and 
Gutierrez, College of Medicine and Surgery, and Doctor Cox, 
interne, Philippine General Hospital. 

4. The entomologic survey, under the direction of Mr. Banks of 
the Bureau of Science, assisted by Mr. Dalburg, of the same 
Bureau, and the mosquito brigade of the San José Estate. 


It is needless to say that all these various sections of work 
were so correlated as to make the study of individual case records 
applicable through all sections of the work. The laboratory sec- 


142 The Philippine Journal of Science 1914 


tion performed blood examinations and stool examinations and 

made hemoglobin estimations. A series of 1,120 employees and 

members of their families were examined clinically by the clinical 

section, with particular reference to the general physical effi- 

ciency, condition of the skin, respiratory system, circulatory 

system, and abdomen, paying particular attention to the “spleen 

index.” The entomologic survey was of a general nature, par- 

ticular attention being paid to the location of breeding places of 

the anopheline mosquitoes and a study of all species of mosquitoes 

possibly concerned in the transmission of malaria. The sanitary 

survey consisted in a systematic investigation of the property of 

the company as follows: 

Population: Men, women, children. 

Births. 

Deaths: Adults, children. 

Medicines: Quinine, quality and method of use; patent medicines, kind and 
quality sold. : 

Food supply: Rice, varieties and quantities used; canned goods; meat; 
greens; chickens and eggs; miscellaneous foods. 

Water supply: Kinds, sources, method of handling, class of consumption. 

Sewage and garbage: Collection and disposal. 

Water-closet facilities. 

General grounds: Stables, manure, etc. 

Houses: Construction, overcrowding, lighting. 

Cooking: Where and how. 

Laundry: Where, how, by whom, frequency of changes. 

Bathing: Facilities and customs. 

Venereal problem. 

House surroundings: Underneath and around; animals. 

Flies and mosquitoes. 

The findings of the various sections of the commission are 
discussed under appropriate headings in the various articles 
making up this report. The last two chapters of the report, 
namely, the summary and conclusions and the recommendations, 
bear the unanimous indorsement of the commission. 


2. GEOGRAPHY AND GEOLOGY 
By F. A. DALBURG 


Situation —The San José Estate is situated in the south- 
western part of Mindoro and on the low plain at the foot of the 
western cordillera system. Mangarin formerly was the principal 
place of habitation, but at present the new town of San José 
has been built near the center of the estate. San José is sur- 
rounded by lowlands which formerly had only a vegetation of 
talahib and buri palms, but is now devoted to the cultivation of 
sugar cane. (Fig. 1.) 


IX, B, 2 Sanitary Survey in Mindoro 143 


Area.—The area of Mindoro is given in the Atlas Filipinas 
as 10,987 square kilometers (3,972 square miles) with a coast 
line of 518 kilometers (322 statute miles). The San José Estate 
contains an area of 22,485 hectares (55,538 acres). 

Drainage.—There are several streams ‘passing through the 
San José Estate of which the two principal ones are the Busu- 
anga and Lumintan (Luminatao, Lumitao) Rivers. These rivers 
flow all the year round, and furnish abundant water for irriga- 
tion purposes. The smaller streams have an intermittent flow 
or form submerged marshes. These marshes during the rainy 
season become lakes, and are the source of numerous smaller 
streams that are dry during the months of no rainfall. 

Routes of travel_—San José is reached from Mangarin Point 
by the Mindoro Company railway. There is a trail leading from 
Mangarin to the town, which was formerly the means of access. 
Very little traveling is done to the north of the town as the 
country is thinly settled. 

Geology.—Most of the plain is alluvial, but shows exposures 
of conglomerate and shales toward the foothills. These in turn 
rest on the igneous rocks which form the base of the mountain 
system. Along the mountain range, white cliffs can be seen 
which evidently are composed of limestone. To the south of the 
estate at Santa Teresa, the natives quarry coralliferous limestone 
for use in the sugar industry. It is stated that about 24 kilo- 
meters north of the estate are oil seeps, and in the mountains 
are numerous caves of limestone. No investigation of the alleged 
oil seeps was made, because of lack of time. 

Soil_—tThe soil on the estate varies from silt to a very clayey 
loam. Considerable portions of the land near the large rivers 
are composed of sand and gravel. In other portions the soil is 
a loamy clay, black to yellow in color. Where the clay material 
predominates, it may be necessary to drain the fields to insure 
a good soil for crops. In the more sandy soil where the retentive 
power is low, irrigation will be necessary to insure sufficient 
water for the cane fields. 

Water supply—The water supply is obtained from artesian 
wells which give a good quantity of potable water. In the 
outlying districts, dug wells are used and also small wells dug 
in the sand of the rivers. 


3. THE GENERAL AND EXTERNAL SANITARY PROBLEM 
By W. E. MuSGRAVE 


The general sanitary problem may be divided into what may 
be termed the external problem and the internal problem. Ob- 


144 The Philippine Journal of Science 1914 


viously, in a community where both malaria and tuberculosis— 
two transmissible and dangerous diseases—have an exceedingly 
high incidence, it will be necessary to control the external con- 
ditions with as much care as is required in dealing with the 
internal problem. 

In the place under investigation, the external problem is a 
tremendous one, and presents some unprecedented phases. The 
eastern boundaries of the property are in the foothills of the Min- 
doro mountains where the wild tribes, the Mangyans, about 
whom very little is known, live, and practically nothing is known 
of the incidence and varieties of infectious diseases among them. 
Fortunately, intercommunication between the employees of the 
corporation and other citizens of the lowlands, on the one hand, 
and the wild men of the mountains, on the other, is rather limited. 
However, due to trade requirements and the increasing attitude 
of friendship, the association between these people is becoming 
more general, and in the course of time this mingling will in- 
fluence the sanitary problem, both of the lowlands and the wild 
tribes of the mountains. Up and down the coast line, north, 
south, and west of the properties of the corporation, there are 
located a number of barrios and villages with the closest asso- 
ciation and intermingling of populations that it is possible to 
obtain. 

THE MUNICIPALITY OF PANDOROCAN 


At the present time, all the part of the east coast of Min- 
doro including the properties of the Mindoro Company and the 
San José Estate is within the municipality of Pandorocan, and 
this municipality, in turn, is under the provincial government 
of Mindoro, which is located at Calapan on the opposite coast of 
the island. 

The geographic outlines of the municipality of Pandorocan 
are shown in fig. 1. It consists of the properties of the Mindoro 
Company and the San José Estate, divided as follows: 


Estimated population. 


Bugsanga Camp No. 1 500 
Lubang Camp No. 2 1,400 
Mindoro Camp No. 3 : 820 
Kaminawit Camp No. 4 100 
Irrigation Canal No. 2 300 
Japanese Camp No. 5 50 
Flowing Camp No. 2 50 


Total 3,200 


; 


IX, B, 2 Sanitary Survey in Mindoro 145 


The following outlying barrios contain practically all the 
remaining population of the municipality. 


Estimated population. 


Pandorocan 85 
Mangarin 280 
Caguray 100 
Santa Teresa (Lalaoigan) 150 
Tlin Island 400 
Calintaan 300 
Bulalacao 200 

Total 1,465 


A sanitary investigation of a number of the barrios has been 
made as follows: 

Mangarin.—With a population of approximately 300 people, 
Mangarin is the oldest inhabited village in Mindoro, being nearly 
three hundred years old. During its existence of these hundreds 
of years, its population has remained practically constant, the 
deaths being just about equal to the births. It is located on a 
low-lying point in Mangarin Bay, is surrounded by swamp 
marshes, and is without drainage. It will be impossible to 
institute drainage without incurring unjustifiable expense. The 
total value of the improvements and real estate in the entire 
town does not exceed 5,000 pesos.: The inhabitants are dirty, 
lazy, and diseased. A medical survey of the population shows 
a “spleen index” among the children of 98 per cent; a clinical 
anzemia index of practically the same figure; a skin-disease index 
of 100 per cent; a very high incidence of tuberculosis and in- 
testinal diseases ; and an infant mortality that is simply appalling. 

After a rather careful examination of this population, it is 
my firm belief that every man, woman, and child of the village 
is infected with malaria, and certainly every one of them has 
scabies, and, in most instances, there is at least one other skin 
disease present. The most practicable way of reaching Mangarin 
from San José is by train to Kaminawit (port of Mangarin), 
a distance of approximately 15 kilometers, and thence by boat 
to the village, a distance of about 2 kilometers. The intercourse 
between the people of this village and San José is constant and 
considerable. The only excuse for the existence of this village 
is that it is convenient to very profitable fishing water. 

Santa Teresa (Lalacigan).—Santa Teresa is a comparatively 
newly constructed village with a very nearly ideal location. It 


*One peso Philippine currency equals 50 cents United States currency. 
One peso equals 100 centavos. 


146 The Philippine Journal of Science 1914 


is on sandy soil in the foothills with plenty of drainage and 
good water supply, and is about 10 kilometers by water from 
Mangarin. At the present time, the “spleen index” among the 
children is 24 per cent, and this malaria incidence is explained 
by a single breeding place of mosquitoes in which many anophe- 
lines were found by Mr. Banks during our investigation. Other- 
wise, the inhabitants of this village are remarkably healthy; 
with the expenditure of a very small sum of money the location 
could be made ideal from a health standpoint for a modern 
sanitary barrio or even a city. 

Toong (estate of the Recoleto Fathers) .—The small barrio of 
Toong is situated in a beautiful valley of the Caguray River, 
and ought to be, with very little attention to sanitary arrange- 
ments, an ideal barrio. As it is now, there are several 
breeding places for anophelines, and the “spleen index” of the 
children is 36 per cent. In one house, containing only one 
room, 18 persons live, eat, and sleep; the cooking is done in 
this same room. There are in this house at the present time 
8 cases of malaria and 2 of tuberculosis; there is 1 child with 
congenital syphilis, and there have been 2 deaths from malaria 
during the past twelve months. 

Caguray.—tThe village of Caguray is situated on the opposite 
bank of the Caguray River and over 1 kilometer southwest of 
Toong. The “spleen index” among the children of this town 
is 80 per cent, and there are numerous breeding places for 
mosquitoes. The location of the village is satisfactory, and with 
very little attention to general sanitary conditions could be 
made a very healthful spot. 

Pandorocan.—Pandorocan is the capital of the municipality 
of the same name. It is located on the property of the Mindoro 
Company, and consists of temporary houses which also are the 
property of the Mindoro Company; the whole place formerly 
was a temporary railroad camp. There is a population of 
between 35 and 50 people with the usual municipal officers, 
including 1 Constabulary officer and 4 Constabulary soldiers. 
One of the soldiers was ill in bed with malarial fever at 
the time of the inspection; one other gave a history of 
chronic malaria, and 3 of the 4 had enlarged spleens. The 
“spleen index” of the entire population is 65 per cent. All of the 
members of one family consisting of father, mother, and 4 
children had malarial fever. There were a number of cases of 
tuberculosis, and the majority of the population are under nour- 
ished and anzmic. 


IX, B, 2 Sanitary Survey in Mindoro 147 


It seems that the location of the capital of this municipality 
at Pandorocan was an accident. It is a deserted railroad camp 
of the Mindoro Company with a few poorly constructed houses 
and a total population of about 35 people, including the municipal 
authorities and 4 Constabulary soldiers. It certainly is not 
located in a particularly healthful spot, neither is it easily acces- 
sible to most of the important barrios or other properties under 
its jurisdiction. 

The entire population of the municipality numbers about 
5,500, of which approximately 3,200 are employees and families 
of the Mindoro Company and San José Estate; the rest of the 
population is distributed among the above-mentioned barrios. 
The income of this municipality is unusually large for one of its 
size, and a vast majority of this revenue comes from the taxes 
paid by the Mindoro properties. At the present time, there are 
about 12,000 pesos in the treasury. The municipality owns no 
real estate and no permanent buildings. The funds of the munic- 
ipality, as well as its ordinances and other legal procedures, are 
under the jurisdiction of the provincial government of Mindoro 
with headquarters at Calapan. 

Detailed information regarding the disposal of municipal 
funds was not investigated, but it is certain that satisfactory 
police and sanitary protection are not given to the inhabitants 
of the municipality, neither are police and sanitary protection nor 
any other benefits furnished the Mindoro Company and San José 
' Estate in return for the large share of the taxation which is 
borne by these corporations. 

The barrios of this municipality are the poorest located, 
poorest cared for in both police and sanitary protection, and are 
the most generally and severely infected villages that I have seen 
in the Philippine Islands. One barrio, namely Santa Teresa, 
of the entire municipality has a satisfactory sanitary location, 
which in this case is all but ideally chosen. Water ‘supply, 
drainage, easy approach by sea with deep water close to the 
shore, sandy soil, and shade trees are the principal advantages 
offered. At a very small expense, all possible breeding places 
for mosquitoes could be destroyed and a very desirable location 
for the municipal capital established. 

It is recommended that Santa Teresa be selected as the capital 
of the municipality and that the headquarters now at Pandoro- 
can be transferred to this site. 

It is further recommended that the barrio of Mangarin be 
transferred to Santa Teresa, for as stated in another place in 
this report satisfactory sanitary conditions in this barrio can- 


148 The Philippine Journal of Science 1914 


not be obtained and maintained with the expenditure of any 
reasonable sum of money. 

In view of the fact that such a large percentage of the rev- 
enue of the municipality is received by taxation of the Mindoro 
properties, it is recommended that satisfactory police protection 
and justice of the peace accommodations be furnished San José 
as a partial return for this revenue. At the present time, the 
municipality governs through ordinances, and in general the 
local officials have very little authority to transact Government 
business in proportion to the requirements of the situation. 
The enormous expenditure of private funds and the rapid growth 
in the population of Pandorocan make it necessary that the 
form of government now in existence in this municipality be 
modified and that it be strengthened to meet the demands of 
the situation. It is assumed that the Government maintains 
the right of proper sanitary and legal supervision of the prop- 
erties of the corporations interested in Mindoro in exactly the 
same way that it retains similar supervision over the private 
properties and estates of other citizens and corporations of the 
country, and the situation at the present time in San José is 
such as to demand serious consideration and more active, aggres- 
sive, and competent discharge of the usual governmental duties 
and privileges granted to citizens of thickly populated districts. 

The health problem in San José and the surrounding barrios is 
such an important one that it should be under the supervision 
of athoroughly competent and well-trained public-health offi- ~ 
cial with all the authority usually delegated to such an officer. 
The corporations interested in the territory under discussion 
already have very large sums of money invested, and con- 
stantly are increasing this investment. It is to their interest 
that the present unsatisfactory health conditions be improved, 
and the management of the companies is showing a very com-- 
mendable spirit in its attempt to improve these conditions. 
However, it is handicapped in many particulars, not the 
least of which is the lack of authority for the enforcement of 
just sanitary laws and regulations among the employees and 
other inhabitants of the property. The Government is inter- 
ested in the improvement of sanitary conditions for the same 
reasons that it is interested in this problem in other parts of the 
country, and the solution of the difficulties at San José is not 
possible except under the combined efforts of the corporations 
and the Government officials under a united directorate. 

From a careful investigation of all phases of this question 
on the ground, I am fully convinced that the corporation in- 


IX, B, 2 Sanitary Survey in Mindoro 149 


terests are ready and anxious to meet the Government in any 
proposition looking to the alleviation of the very unsatisfactory 
conditions existing at the present time, and I recommend 
and urge that the Government take up this matter on a broad 
basis, reach a line of agreement for methods of work, and pro- 
ceed to clean up the area in question and remove the stigma 
of the “white man’s grave” from one of the richest and most 
attractive territories of the Philippine Islands. Definite recom- 
mendations regarding this point will be found in another place 
of this report. 


4. THE INTERNAL SANITARY PROBLEM 
By THOMAS W. JACKSON 


As a member of the Mindoro Malaria Commission, represent- 
ing the Bureau of Health, I made certain observations, under 
the direction of the chairman of the commission, relating to 
various phases of the health problem at the San José Estate 
and adjacent properties. These observations may be grouped 
under the general title, The internal sanitary problem. Con- 
sideration was given in this investigation to the following sub- 
jects: (1) Topography and climate; (2) population, individually 
and collectively, from a medical standpoint; (3) housing con- 
ditions and nutrition; (4) the sanitary and medical organi- 
zation; and (5) a study of the causes for the high morbidity 
and mortality rates at San José. 


I. TOPOGRAPHY AND CLIMATE 


The San José estate is situated on the low plain that extends 
from the coast to the foothills of the mountains in southwestern 
Mindoro. (Fig. 1.) 

This plain is intersected with numerous arroyos which contain 
flowing water at all times and which in the rainy season become 
deep and torrent-like streams. The two great rivers are 
the Busuanga and the Lumintan, flowing in somewhat parallel 
directions from the mountains to the sea and distant about 12 
kilometers from each other. The Lumintan is the most north- 
western, and crosses a portion of the sugar estates, while the 
Busuanga passes directly through and across the estates within 
1.5 kilometers of the sugar mill. Magbando River, a smaller 
parallel stream, flows close to the sugar mill, and with its 
branches provides a running-water system through the “Min- 
doro” and “Lubang” camps, and furnishes as well a natural 
irrigation system. .The banks and bed of this stream have been 


150 The Philippine Journal of Science 1914 


the scene of much clearing during the past year as an anti- 
mosquito measure. 

Talahib grass covers the greater part of this plain, growing 
to a height of 4.5 meters in some places. Naturally, it furnishes 
excellent shelter for mosquitoes, especially in the rainy season, 


2 
“© LeGeno 
© Mindoro Co. Comp 
@) San José Co, Camp 
© Hospital 
@) Sugar Mill 
© Garbage and Sewage Disposal Pits 
© Cemetery 
@ Planted Fields 
Ploughed Fields 
: © Artesian Well 
THE TOWNSITE OF 9 _--=---—"s, ‘ Wireless Station 


SAN JOSE ESTAT “| © tuarters for oMtcials 


MINDORO ‘ , \ @ Stables 
SCALE ‘ ® Offices and Store Houses 


Fic. 2. The townsite of San José Estate, Mindoro. 


when the streams overflow their banks and inundate the entire 
tract. Talahib grass, however, is something of an index of the 
suitability of the soil for sugar culture; therefore, it may be 
expected eventually to disappear entirely before the steam plows 
of the sugar planter, which have already converted 616 hectares 
of talahib-covered area into sugar-cane fields. 


ii 


IX, B, 2 Sanitary Survey in Mindoro 151 


An extensive and elaborate system of ditching and draining 
has been undertaken and is going forward under the direction 
of the engineers. This cannot fail greatly to facilitate the control 
of mosquito breeding, just as the disappearance of talahib 
deprives the mosquitoes of their natural shelter. The increased 
porosity of the soil from cultivation should be of great assistance 
in lessening the number of mosquito-breeding places. 

The climate of San José does not differ materially from that 
of other portions of the Philippines of similar latitude (12° 
north), and as the altitude of this plain in few places exceeds 
15 meters above sea level no extremely cool weather is to be 
expected, but sea breezes during the southwest monsoon meet 
with no obstruction and temper the climate pleasantly. During 
three of the six nights spent at San José, comfort demanded a 
heavy blanket or two light ones. Upon the other three nights, 
a very light covering sufficed. 

There is nothing in the climate, in itself, to account for the 
unhealthfulness of this region. The vegetation and rainfall 
combine to provide breeding places for mosquitoes, and in 
this indirect way only may the climate justly be held responsible 
for malaria. With extensive cultivation and drainage, we should 
expect the ultimate passing of the mosquito and the repetition 
of the history of malaria in the lowlands and river bottoms of 
the central United States, once notorious by reason of the so- 
called paludial fevers, but now healthful and providing wealth 
and sustenance to the millions of prosperous people resident 
there. 


II. POPULATION, INDIVIDUALLY AND COLLECTIVELY, FROM 
A MEDICAL STANDPOINT 


In the matter of composition, the population may be described 
as heterogeneous; while the Filipinos predominate largely, 
Japanese, Chinese, Americans, Hindus, and Negroes are also 
present. The Japanese are the second strongest people numeric- 
ally, while the Chinese and Negroes are very few in numbers. 
The population of San José, comprising the three camps of 
Lubang, San José, and Mindoro, for the quarter ending Decem- 
ber 31, 1912, was given in the official health report of the surgeon 
to the company as 2,800, while that of the third quarter of the 
year, ending September 30, was given as 3,285. An average 
population of 3,000 people may, therefore, be assumed. It is 
ultimately proposed to employ and quarter 10,000 persons, ac- 
cording to the statement of company officials. 


152 The Philippine Journal of Science 1914 


The exact influence of race and nationality upon the resistance 
of this populace to disease cannot be estimated, and even ap- 
proximate estimates would be of little value. 

It is my opinion, based upon physical examinations, that the 
American comes first in physical fitness, with the Japanese 
second. The Filipino is more subnormal in development than 
any of the other nationalities, and next higher, but not greatly 
above him in the physical scale, comes the East Indian. The 
habits of life and of religion, in the case of the Hindus, may play 
some part in the determination of their resistance to disease, 
but no generalizations are possible. 

The tractability of the various nationalities also enters into 
the problem, and in this regard I believe the Filipino, under 
proper authority, to be fully as tractable and amenable to sanitary 
and medical restraints and discipline as any of the other races 
represented here. I emphasize the matter of authority, in as 
much as it is absolutely vital to the solution of this problem, and 
at present it seems to be the most neglected part of the sanitary 
and medical scheme. 

Concerning the matter of resistance to the sun’s heat and light, 
we have no reliable data, and the present confusion and disagree- 
ment among students as to the effects of tropical light upon man 
warrant us in devoting ourselves to a search for more tangible 
and evident causes of disease. 

As to the general physical condition of the working populace 
at San José at the present time, it may be said that the people 
are poorly nurtured and anemic. Many are harboring malaria 
parasites, and are suffering from the blood-destroying activities 
of these organisms. Considering the character of the work done 
and the hours of labor, all are underfed and are thereby rendered 
less resistant to invasion by most of the infectious diseases. A 
good many are tuberculous and consequently unfit for service of 
any kind. The term “semimiserable” does not understate the 
physical condition of the people in general. 

I am unable to give the proportion of women and children to 
the 3,000 inhabitants, but it is certainly much less than that 
normally existing in Filipino villages. The adult population 
furnishes the greater portion of the sickness. 

Of the deaths occurring during the last quarter of 1912, 
18 per cent were among children, whereas in Manila the constant 
percentage of deaths among children exceeds 50 per cent of the 
whole. 

During the two months immediately preceding our visit, 10 


IX, B, 2 Sanitary Survey in Mindoro 153 


children were born, twice the number which died in the final 
three months of 1912. No previous records of births were 
obtained. 


Til. HOUSING CONDITIONS AND NUTRITION 


Housing conditions.—As the sugar companies are endeavoring 
to work out the question of proper housing, I will only say that, 
in my opinion, the barrack system now in use is defective in 
many ways and that the individual (family) house seems to 
promise better results generally. The condition of overcrowding 
and of insanitary interior partition of barracks into small rooms 
exists, and needs correction badly. 

Much trouble, in my opinion, might have been avoided, had 
the companies exercised their undoubted right to regulate interior 
as well as exterior conditions about their houses. Until such time 
as they do so, no system of house building will remedy the un- 
wholesome conditions which exist. That the companies can limit 
the number of persons per house; exclude chickens and animals; 
prevent the unauthorized construction of partitions, sleeping 
closets, and sleeping shelves in their buildings; oversee ventila- 
tion; and insist upon the maintenance of screens and upon 
the use of the sanitary appliances provided by them is as plainly 
possible as it is for them to regulate outdoor conditions. 

Nutrition —The important matter of nutrition can only be 
dealt with here in a general way. The fact of general malnu- 
trition or subnutrition among these people was abundantly 
established by the commission. 

The methods of supplying food for the laborers and their 
families were investigated, and as a result the fact that the food 
is sold at an abnormally high price was established. Members 
of the commission priced the staple articles of food on sale at the 
company stores and at tiendas, and found that prices were decid- 
edly higher than those prevailing throughout the Philippines, and 
in many cases exceedingly high. 

The messes of the restaurants conducted by the companies were 
also investigated, and it was found that three classes of meals 
were provided at rates of 20, 15, and 10 pesos per month, respec- 
tively for first-, second-, and third-class board. In the opinions 
of the chairman of the commission and myself, only the first-class 
board, at 20 pesos per month, is suitable to subsist a workman 
engaged at hard labor on the sugar estates. From inquiry of 
the mess managers, it was found that less than 5 per cent of 
the boarding laborers selected this fare. A larger percentage 

124316——2 


154 The Philippine Journal of Science 1914 


paid 15 pesos for second-class board, but the great majority 
was found to be living on the 10-peso fare, which is an absolutely 
inadequate ration from either the viewpoint of caloric value 
or that of hunger satisfaction. 

We need only to state the fact that the monthly wage of the 
laborer amounts to 26 pesos (for twenty-six working days), to 
make clear the reason why practically none of the men select 
the first-class fare. The situation in the case of the families 
providing and preparing their own food is less easily studied, 
but with the supplies at hand and the high prices demanded 
for necessities it cannot fail to be most unsatisfactory. 

The remedies for these conditions are obvious, and I am of 
the opinion, as I have stated before, that the sugar companies 
are disposed to remedy evils brought to their attention. There 
must be provided for the people an abundant and constant supply 
of good food at fair prices. There should be vegetables, meats, 
and fish at all times. Excellent fish are obtainable close at hand, 
and vegetables may be grown without difficulty. Pigs and cattle 
should be raised and sold at prices within the reach of the popu- 
lace, while chicken raising should be practiced and encouraged. 
There is no insuperable difficulty in the way of providing 
good food at living prices and in constant abundance, but some- 
thing more than haphazard management and good intention is 
necessary to remove this real and regrettable state of affairs. 

The relation of this nutrition question to health maintenance 
is sufficiently clear, and demands no argument. Without ques- 
tion, diminished resistance by reason of underfeeding is a factor 
entering into the matter of recovery from most infections, even 
if it cannot actually be shown to be a factor in the matter of 
infection itself. The fact that we have at San José, to start 
with, a class of workmen, underfed and deficient in stamina, only 
emphasizes the obligation on the part of those who engaged them, 
and in many cases imported them, to correct this underfeeding 
and to care for them physically in every necessary way. Of 
course, there is no reason why men should be retained in em- 
ployment or maintained if they are intractable and if they resist 
sanitary and medical measures. It is extremely likely that many 
had malarial, hookworm, and tubercular infections when they 
arrived and that their physical woes were not at all acquired 
in Mindoro, but since they were accepted as workmen without 
physical examination they must now be properly fed and cared 
for or returned to their homes, regardless of the way they lived 
before accepting employment and residence in Mindoro. 


——————— lll Oe 


a ee ae ee se 


——_— = 


1X, B, 2 Sanitary Survey in Mindoro 155 


IV. SANITARY AND MEDICAL ORGANIZATION 


To establish the fact that the mortality and morbidity rates 
are actually high, let me quote briefly a few statistical notations 
from the San José hospital records. 

Average population for 1912, 3,000. Total number of deaths 
from July 1, 1912, to January 18, 1913, 182. (If this rate for 
seven months were maintained for the entire year, the deaths 
would reach the number of 216, a death rate of 73 per thousand 
of population, or 7.3 per cent.) Asa matter of fact, the monthly 
death rate has varied considerably. During the month of July, 
1912, it exceeded 200 per thousand annually (statement of the 
company’s surgeon). The death record of the periods previous 
to July 1, 1912, was not furnished. 

The causes of death were obtainable in only the last 34 cases 
recorded (October 5, 1912, to January 18, 1913), and are as 
follows: 


Malaria? 18 
Beriberi 3 
Pneumonia and tuberculosis 6 
Premature birth 3 
Ameebic dysentery and enteritis 2 
All other causes 2 

Total 84 
Per cent of all deaths due to malaria 52 


For the 98 deaths recorded from July 1 to October 5, 1913, 
diagnoses are recorded for but 34, of which 16 (or 47 per cent 
of the whole) are ascribed to malaria. 

No numerical record of cases of malaria treated in hospital 
was furnished me, but if the record of deaths is even approxi- 
mately correct a tremendous incidence may be inferred, if one 
has in mind the comparative rarity of death from treated 
malaria, even in tropical countries. For example, in 1899, only 
4 deaths occurred among 1,904 infected persons actively treated 
at Pinar del Rio, Cuba. 

When completed, the examination record of more than 1,100 
blood smears taken from the people at San José will give a fair 
index of the degree of malarial infection here, but it must be 
remembered, of course, that not every infected individual ex- 
amined will show parasites in the single specimen taken, espe- 
cially as many of the infections are with the estivo-autumnal 


* All deaths in which the records designate malaria as one of the causes 
of death are included. 


156 The Philippine Journal of Science 1914 


parasite, which in certain phases of its human cycle is practically 
absent from the peripheral blood circulation. 

As for the so-called “splenic index,” I share the belief held by 
a number of tropical workers that it is a very fallible criterion, 
especially in the presence of other anemia-producing conditions, 
such as those which prevail at San José. It has a certain value, 
however, and so far as it goes it gives support to the clinical 
findings of malaria in San José. 

In 1911, Schapiro reported a general splenic index of 17.7 per 
cent at San José, based upon an examination of 333 persons. 
For those under 16 years of age he found the splenic index to 
be 37+ per cent. From the same report we learn that in 
January, 1911, the estimated mortality rate per annum, based 
upon the month’s deaths, was 88 per thousand, or 8.8 per cent. 

The matter of the physical condition of the inhabitants of the 
near-by towns, while interesting and undoubtedly bearing 
considerable weight upon the problem before us, is less im- 
mediately important than is generally supposed, and it is cer- 
tainly less so than the cure of the sick at San José. At San José, 
we have a present population of 3,000 persons, a large per- 
centage of whom are infected with malaria. The combined 
population of the other towns and barrios of southwestern Min- 
doro, within a radius of 24 kilometers, does not approach this 
total. We have also to bear in mind the fact that to convey 
malaria to the San José sugar estates the human malaria carriers 
from these settlements must visit San José and encounter anophe- 
line mosquitoes, or that laborers, women, or children from the 
sugar camps must visit the infected barrios and encounter in- 
fected anophelines. 

The company’s surgeon believes, not without some apparent 
reason, that, given complete control of the anopheline mosquitoes 
at San José, a condition which he believes to be actually attain- 
able and even now measurably accomplished, the visiting malaria 
carriers will not constitute a menace to the inhabitants. He 
then needs only to fear that the uninfected people from San 
José will visit the barrios and towns and there acquire malaria, 
and he needs only to safeguard the members of this traveling 
population during their excursions and observe them after their 
return, treating such as acquire malaria while absent from the 
estates. 

Theoretically sound, at least in part, this proposition appears 
to me to be absolutely impracticable of application and bound 
to result disastrously for several reasons. 

One of these reasons is the impracticability of ever, or at 


IX, B, 2 Sanitary Survey in Mindoro 157 


least within some years, securing such ideal mosquito-free con- 
ditions throughout the company’s settlements as is contemplated. 
I consider this attainment improbable, in spite of the present 
dry-season indications and the admitted extreme scarcity of 
mosquitoes at San José. The rainy-season test has yet to be 
applied. Pending the results of this test, I confess that I am 
unconvinced of the absolute and permanent eradication of an- 
ophelines at San José. — 

Another reason is the impracticability of actually controlling 
the laborers and others, who visit infected barrios, during their 
absence from the estate and upon their return. 

With an abundance of malaria carriers resident at San José— 
hundreds in fact—and a few anophelines to act as vectors, there 
is every reason to expect the disease to continue indefinitely at 
the sugar estates unless more active therapeutic measures be 
instituted. 

In 1911,? antimalarial recommendations were made for a 
drainage system that would entirely exclude mosquito life; for 
the leveling, grading, and drainage of the town site; and for 
the clearing of vegetation from the banks of the Magbando 
River and for clearing its channel and straightening its course. 

These recommendations have been carried out so thoroughly 
that to-day the Magbando River at San José is an open, un- 
obstructed waterway with banks practically denuded and offering 
absolutely no opportunity to the mosquito seeking a breeding 
place. The elaborate drainage system already installed and now 
undergoing construction, with cement gutters and drains and 
with straight deep ditch sides and proper grades, are indications 
of great effort and expenditure. The buildings are arranged 
symmetrically and in order in well-policed streets. All of these 
things speak in praise of the energy of the company’s surgeon, 
the company’s engineers, and the management which has supplied 
the large sums of money required. Mosquitoes at the time of 
our visit in January, 1913, were extremely hard to find. Yet 
malaria continues to exact a terrible toll in life and health. 

Granting that some malaria is constantly introduced from the 
infected village of Mangarin, I am still unable to rid myself 
of the impression that the great majority of the fever cases 
occurring at San José are in “repeaters.’”’ The medical officers 
of the company stated to me that a very high percentage of the 
cases were recurrences. From inquiry, observation, and ex- 
perience I know that the average length of time spent by malaria 


* Annual Rep. P. I. Bur. Hlth. (1911), 48. 


158 The Philippine Journal of Science 1914 


patients in the hospital—from two to three days—is far short 
of the time required to cure cases of malaria. Furthermore, 
the routine treatment administered is not such as to lead me to 
expect any permanent cures, and with the lack of systematic 
after-treatment it would be strange, indeed, if the majority of the 
cases did not recur. 

So far as the immediate problem is concerned then, I consider 
it a medical rather than a sanitary problem. 

The following experiences caused me to feel sure of that which 
I had strongly suspected; namely, that many cases never come 
under the observation of a physician at all and do not appear on 
record anywhere, officially or otherwise. 

As a member of the commission, one of the duties apportioned 
to me was that of making a house-to-house inspection of the 
three camps, Lubang, San José, and Mindoro. These inspections 
were made at various times. 

On January 14, at about 2 p. m., I visited the most distant 
part of San José camp, extending along the Busuanga River bank. 
In the 5 houses nearest the end of the river row, I found 7 persons 
suffering with fever. Blood smears for examination were taken 
from all, and directions to visit the hospital were given. Only 
one person did so. Upon inspection of the same houses on the 
morning of January 15, I found that 6 of the 7 had returned to 
work. Upon the second visit, 5 new cases were encountered— 
all cases of malarial fever. None of these individuals had ap- 
plied for treatment or was receiving it, and none is officially on 
record except the one who was persuaded to visit the hospital. 
These cases from San José included both simple tertian and 
malignant tertian (zstivo-autumnal) cases, and parasites were 
found without difficulty. 

On January 17, while inspecting a cookhouse in the Mindoro 
camp, the head cook was found shaking with a chill. His blood 
contained malaria parasites. 

On January 17, at 2 p. m., I visited a row of houses (4 
barracks) in Lubang camp, and found 18 cases and took blood 
specimens from all. 

The rest of the barracks, 22 in number, were then imme- 
diately inspected by another member of the commission (at 3 
p. m.), who reported that about 50 persons were at home with 
fever at the time of his visit. About 30 blood smears were 
taken. Men, women, and children were included in this number 
of persons. None of the fever cases was “officially” sick, and 
none of the 26 cases casually discovered by me was under 
treatment. 


IX, B, 2 Sanitary Survey in Mindoro 159 


A suggestive circumstance connected with my inspection was 
the surprise expressed by the sanitary policeman, who accom- 
panied me through the Lubang camp buildings, that persons with 
“only a little fever’? should require hospital treatment. He ex- 
pressed the opinion that they ‘‘would all be at work to-morrow.” 

The following “condensed dispensary reports” were furnished 
at the San José Hospital in the morning of our return to 
Manila. I introduce them just as transcribed from the records. 


TABLE I.—Condensed dispensary reports for week ending January 11, 1918. 


A Con- 3 F 
Febric-| - - | Acute| Diar- | Surgi- 
ula. cab coryza.| rhoea.| cal. Total. | 


Lubang (camp 1); population, 1, 400: 


\ CHT (GIT ees aps Aon ae oes ae ie es 34 1 6 5 25 71 

Days'lost from work 2252.5 2.-2. 2522-5220 34 1 7 6 31 79 

Capen gent tOMmoAn ha bss fe Ser cp se - ede Ne e ne 5 
Bugsanga (camp 2); population, 400: 

Cape esata el Se 2 ae 10) | 5 [hl eee eee Poe easy 11 

Days lost from work -......---..-----.---- n'y (Ee eae Se ANE SS ae eae Se 19 


Gases seniito Nospital eee ee he es tf 
Mindoro (camp 3); population, 720: 


(0212): aaa Rs ee od i eR SX eects 4 5 16 62 
Days lost from work _-__------------------- Sy} ee ees 6 6 23 74 
Gardesisentito Nospitalo ase foo ke See ee ee ae 5 


Kaminawit (camp 4) (port of Mangarin); popu- 
lation, 80: 


AGAR Crs eee 30s NA TS ah ES SSS re re | 8 a wf es al ge ek 

UT STEN ICGT TU Gd Shae aes ae ORE OC Pee Sa a ee Seer! eee eee eee SNe (ae Rie 

AP I RCYAPALCH OSD IL eee eee eee ee ee eee ee ocean 1 
Irrigation canal (camp 2); population, 200: 

(Ot: C(O ee ar ne Pewee ee Si hee 6 1 10 25 

Days. lost: from work |... =2..---2.-~---4_. Sots ae 6 1 12 27 


GAREoInEN tic NOS DLA oor oe eneres ese se ee eee eae aerate 5 


Grand total dispensary service: Population, 2,800; patients, 169; days 
lost, 199. 
Signed by physician in charge. 


A striking omission from the list of diseases is that of 
malaria. Another is the fact that 99 out of 169 cases were cases 
of “‘febricula,” a term practically banished from modern medical 
nomenclature. Substitute the term “malaria” for “febricula,” 
and we have 58 per cent of all cases treated in dispensary due 
to malaria. The days lost from this cause, although averaging 
but one day per case, amount to 50 per cent of the total. 

Concerning the so-called dispensary service, it seems to con- 
sist chiefly of a practicante who visits cases at their houses 


160 The Philippine Journal of Science 1914 


and administers medicine. He may also send cases to the 
hospital, but as indicated by the above reports and by the com- 
mission’s investigations he does so with comparative rarity. 
The clinical thermometer may be a familiar diagnostic instru- 
ment to him, but the clinical microscope apparently is not. This 
service should scarcely be dignified with the name “dispensary.” 


A STUDY OF THE CAUSES OF THE HIGH MORBIDITY AND 
MORTALITY RATES AT SAN JOSE 


Doctor Schapiro of the Bureau of Health gave as the cause 
of sickness, at the time of his inspection in 1911, the following 
9 factors: (1) Long hours of work; (2) overcrowding in hot 
houses; (8) low nourishment in a poor class of laborers; (4) 
unsafe water supply; (5) lack of mosquito protection; (6) de- 
layed medical attention; (7) poor hospital facilities; (8) no 
sanitary control of garbage and feces collection; and (9) fly 
propagation in the corral, strewn garbage, and feces about 
residences. 

With regard to (1) long working hours, (2) overcrowding 
in hot houses, and (3) low nourishment in a poor class of 
laborers, it may be said that all of these causes still operate, 
although some attention has been given to all of them by the 
companies. (4) Unsafe water and (5) lack of mosquito pro- 
tection have received a great deal of attention, and are no longer 
active factors of sickness, except that individual mosquito pro- 
tection is not practiced. However, a square kilometer and a 
half of territory have been practically freed from mosquitoes, 
and this, it must be admitted, is an accomplishment of great 
value. The artesian wells and distribution system have com- 
pletely and satisfactorily solved the water question. Supply, 
quality, and service are highly satisfactory for present needs. 
(6) Delayed medical attention—not due to fault upon the part 
of medical officers, however—and (7) poor hospital facilities 
still continue to operate in the manner I have pointed out else- 
where in this report. By the reforms inaugurated and operated 
by the companies, factors (8), garbage and feces collection, 
and (9), fly propagation and soil pollution by garbage and feces, 
have ceased to operate. This does not mean that breakdowns 
in the machinery of the sanitary system never occur. On the 
contrary, I think that they are rather frequent in occurrence. 
The important thing is the fact that a system of collection and 
disposal has been installed and will eliminate the dangers from 
those wastes and insects if properly supervised. 


IX, B, 2 Sanitary Survey in Mindoro 161 


Having in mind the most important cause of sickness at San 
José, malaria, the following factors deserve special consideration: 
(1) Lack of disposition on the part of the infected people to 
seek medical attendance; (2) lack of authority on the part of 
the doctors and company officials to compel hospitalization of 
the sick and adequate after-treatment; (3) inadequate hospital 
facilities and medical forces; and (4) lack of confidence on the 
part of the medical staff of its ability to cure malaria and to 
maintain medical surveillance and control over out-patients. 

1. Lack of disposition on the part of the people to seek medical 
attendance.—The attitude may be explained in part by native 
superstition and ignorance, together with dread of the routine 
method of medication for malaria practiced at the hospital; 
namely, iced baths and hypodermatic injections. A significant 
matter in connection with this lack of disposition to go to the 
hospital is the fact that upon admission to the hospital the pay 
of the employee stops. 

The remedies for this condition of apathy are (a) education 
(entirely too slow for our present purpose), (b) compulsory 
hospitalization, (c) modification of the routine treatment so as 
to be less terrifying to the superstitious native, and (d) con- 
tinuance of pay of employees while in the hospital. 

2. Lack of authority, on the part of the doctors and company 
officials, to compel attendance at the hospital by the sick, and lack 
of adequate after-treatment.—It must be admitted that this con- 
dition seems to be a purely fictitious one. It is difficult to under- 
stand why a vast corporation, owning its land and villages; 
controlling transportation to and from its own estates; furnish- 
ing houses, rent free, to its employees; maintaining timekeepers, 
overseers, sanitary policemen, practicantes, and a hospital sys- 
tem, is powerless to enforce the proper treatment of these 
laborers when they are sick. 

Yet I have for my authority the specific statement of the 
company’s surgeon that he is entirely unable to enforce 
proper hospitalization and out-patient treatment. In reply to 
my question, “What means of enforcing hospital attendance of 
your laborers and detention for a suitable period of time have 
you?” he answered, “Absolutely none.”’ One can only explain this 
alleged condition by two alternative hypotheses. One is, that 
the corporation fears to lose some of its laborers by insisting 
upon proper medical treatment and that it prefers to maintain 
the number at a certain figure, even though many are diseased 
and inefficient (for it must, of course, be aware that efficiency 


162 The Philippine Journal of Science 1914 


goes only with sound, healthy workmen). The second hypo- 
thesis, and the one I choose to accept, is simply this: Neither 
of the companies concerned realizes that it is actually clothed 
with absolute control over the laborers, by virtue of the discharge 
and exclusion power; nor do the companies realize the true ser- 
iousness of the situation and the extent of malarial infection and 
invalidism. This last impression is emphasized by the repeated 
statement of company officials that “in spite of malaria” the 
sugar camps are as healthful as any villages of similar size in 
the Philippine Islands. 

The study of the commission absolutely negatives this con- 
tention, and the death rates settle the matter beyond question; 
there is no need to dwell longer upon this point. 

I have no hesitancy in stating that less than one-half of the 
people who suffer from acute malaria at San José come under 
the observation or treatment of the medical officers. These un- 
treated individuals cannot fail to perpetuate the disease in the 
sugar camps. 

3. Inadequate hospital facilities and medical forces.—There is 
evident need of another hospital at San José. The present plan 
of treating surgical cases, diarrhoeal diseases, skin diseases, 
tuberculosis, and malaria in common wards is a poor one, to say 
the least, and the present hospital is badly crowded. Floor space 
and cubic-air space at least twice as great as the present hos- 
pital offers are needed for the present service and, as I have 
already indicated, the number of patients treated in hospital and 
for the adequate period of detention should be doubled. Malaria 
cases and tuberculosis cases should be separately housed in well- 
screened buildings equipped with modern facilities. Elaborate 
and expensive buildings and apparatus are unnecessary; but the 
hospitals should be of ample size, comfortable, and well arranged, 
having in view utility and modern sanitary administration. A 
laboratory room should be provided. 

The medical force should be doubled at least. The addition of 
another medical officer, who should be a clinician and a microsco- 
pist, is absolutely necessary if good work is to be done. It is 
manifestly impossible for the present staff to handle the hospital 
administration, the clinical material at hand, the laboratory 
work and the out-door sanitary supervision, and antimosquito 
work required at present. Then, too, the demands will be 
greater, rather than less, for some time to come if the general 
plan of hospitalization of all the sick be carried out, as I strongly 
recommend. I can see no escape from this conclusion. Imagine 


——— 


f 
; 
‘‘ 
) 

q 


IX, B, 2 Sanitary Survey in Mindoro 1638 


Fort William McKinley, near Manila, where a similar number 
of persons are cared for under vastly more favorable conditions, 
provided with a medical staff of one American doctor and one 
Filipino assistant, aided by one native trained nurse. While the 
needs of Fort William McKinley, with its staff of ten or more 
surgeons, are not to be fairly compared with those of the San 
José Estate, death rates and sick rates for a similar population 
(numerically) are bound to be compared. The right to live and 
to receive proper care in time of sickness is a common one, and 
is an obligation equally binding upon governments and business 
corporations. 

4, Lack of confidence, on the part of the medical staff, in its 
ability to cure malaria.—With regard to the curability of malaria, 
I can only express my regret that such a lack of confidence 
exists. I have become too thoroughly convinced by experience in 
hundreds of cases of malaria of all kinds, studied clinically and 
microscopically for a long period of time, that malaria is a 
perfectly curable disease—although not always an easily curable 
one—to have this confidence shaken by the present apparent 
failures, especially in the light of the conditions of treatment 
existing at San José. ; 


5. THE MOSQUITO SURVEY IN AND NEAR SAN JOSE 
By CHARLES S. BANKS 


The region comprised in this brief survey is, like many others, 


' namely, Manila, Iloilo, Olongapo, Subig, and Cavite, ideal with 


respect to the conditions under which the malaria mosquito can 
breed. The coast being nearly flat, tidal swamps and esteros 
cut far inland and thereby create an enormous area of semi- 
stagnant salt marsh, in which, unaffected by sudden and com- 
plete tidal washings, alge abound and mosquito larve find ample 
breeding and feeding grounds. 

By reference to the accompanying map, it will be seen that all 
the localities mentioned lie near or on the zone of tidal swamps 
and the people are therefore placed under conditions which are 
perfect from the standpoint of exposure to mosquito attacks. 
It is likewise highly probable that those who were born or have 
lived for long periods of time in these localities have become 
permanent malaria carriers; and it is as a result only of the 
recent attempts to import laborers from other more or less 
malaria-free regions and the consequent infection of nonim- 
munes with acute symptoms that attention has been again 
strongly directed to this locality as a malarial region. 


164 The Philippine Journal of Science 1914 


Since the establishment of the permanent settlements of the 
San José Estate and the Mindoro Company at San José, the 
outbreaks of malaria have been so frequent and of such a per- 
nicious type, both among officials and laborers, that the companies 
have found it very much to their advantage to take active steps 
to eradicate mosquitoes within the inhabited area of their planta- 
tions and to screen the houses of those employees intelligent 
enough to keep screens in repair and screen doors properly closed. 

The area shown on the map, inclosed within solid red lines, 
has been practically freed from both anophelines and culicines 
by the process of drainage and oiling, but outside this area there 
are breeding places of both Myzomyia rossii and Culex ludlowii, 
the two forms being usually associated in the same breeding 
places. (Plate I.) 

Busuanga River irrigation site —This meandering river is an 
ideal place for the best development of Myzomyia and Culex, it 
being, at the time of my visit, somewhat low and its valley 
thickly dotted with isolated pools in which alge grew most 
luxuriantly and mosquito larve bred abundantly. 

Enough larve of Myzomyia were found in the intake site of 
-the company’s irrigation ditch to keep the colony in a con- 
tinuous state of malarial infection. Indeed, several Filipinos 
and Americans who had been sleeping for a number of weeks 
at this place were in hospital suffering from severe malaria at 
the time of my visit. 

The same abundance of anopheline larve was noted at the 
Magbando Camp, some 3 kilometers southwest of San José, and 
where the river had during 1912 carried away about 3 hectares 
of sugar cane. 

Railroad.—The entire line of the railroad, from the wharf at 
the port of Mangarin to the line limit for antimosquito measures 
at San José, lies first through tidal swamp land and then through 
land containing creeks, streams, and more or less stagnant ditches 
and ponds where Myzomyia larve were found without exception 
and in abundance. 

The ease with which mosquitoes can be carried from this 
region to the town of San José will be more apparent when it 
is stated that in a trip made after nightfall from Mangarin 
wharf to San José, in one of the open passenger cars of the 
company, I captured at least 3 females of Myzomyia rossii 
attempting to bite me, who was one of at least 30 passengers. 
It might also be said that in the whole trip of 15 kilometers 
which consumed just seventy minutes, but two stops were made. 
There was absolutely no breeze at this time. 


IX, B, 2 Sanitary Survey in Mindoro 165 


Amindan River.—Amindan River, a small creek southeast of 
San José, was found infested with Myzomyia and Culex larve 
as shown in the map. 

Mangarin.—The village of Mangarin from the standpoint of 
insanitary location could probably not be excelled anywhere in 
the Philippine Islands. This is a well-laid-out town 300 years 
old, with fairly clean streets and well-maintained fences, together 
with houses somewhat above the average of construction and 
cleanliness, set down in a tidal swamp. The effects of the 
proximity of mosquito-breeding places are shown by the ap- 
palling splenic index among the children of 98 per cent and an 
anzmia among the adult population which was absolutely general. 

It is needless to state that malaria mosquitoes were found in 
very great abundance in and around this place and that the only 
possible solution of the malaria problem here would be to abandon 
the place completely. 

This single village is an incubator where the germs of the 
disease are maintained year after year to be sent out in ambulat- 
ing cultures from which every anopheline that bites them can 
draw an abundant supply of the organisms of pernicious malaria 
for the infection of other persons. 

Santa Teresa.—Santa Teresa, situated on Lalaoigan sandspit, 
about 10 kilometers from Mangarin wharf, and having about 
150 inhabitants, is fairly free from malaria mosquitoes; but 
outside of the town, along an ill-defined estero or swamp area, 
were to be found breeding places of both Myzomyia rossii and 
Culex ludlowii with limited numbers of larve at the time of my 
visit. 

This town seemed to be the most promising from the stand- 
point of ease of sanitation of all the towns seen in this portion 
of Mindoro. 

Toong.—About 13 kilometers from Mangarin wharf and 6 
kilometers from the mouth of the Caguray River is situated the 
barrio or rancheria of Toong belonging to the Recoleto Fathers 
and used as headquarters for those laborers who herd the cattle 
belonging to this order. The ground is generally high and fairly 
well drained, but along the river banks are to be found breeding 
places of Myzomyia in fair abundance. 

Children in Santa Teresa and in Toong, examined by Doctor 
Musgrave, showed spleen enlargement indicative of chronic 
malaria infection. 

Caguray.—Caguray, a small, well-laid-out, and fairly clean 
town, is 4 kilometers from the mouth of the river of the same 


166 The Philippine Journal of Science 1914 


name and 11 kilometers from Mangarin wharf. It has the 
same general topography as Toong and the same conditions 
favorable to the breeding of Myzomyia. Larve were found, not 
in abundance, but the spleen index of 44 per cent, higher than 
that of Toong which is only 36 per cent, indicates that at certain 
periods in the year mosquitoes are more abundant than at the 
time of my visit. 

I have no hesitancy in asserting that not a single town, barrio, 
or settlement in the whole region within easy traveling distance 
by small boats of the San José estate is a desirable one from 
which to draw laborers or from which to allow visitors to the 
Mindoro plantation, on account of the great probability that 
they will be malaria carriers and that a little relaxation on the 
part of those charged with keeping the “living area” of San 
José free from Myzomyia rossi will immediately result in these 
laborers or visitors serving as a source of infection to employees 
brought from other places in the Islands. That San José and its 
immediate environment can be freed and kept free from mos- 
quito infection is within the range of possibility, but only at the 
cost of unceasing vigilance and an absolute codperation between 
the engineers in charge of the field operations and the sanitary 
officer and his assistants. 


CONCLUSIONS 


The site for San José was ill chosen, as is admitted by all 
concerned, but such an enormous outlay of money has been made 
by the companies interested that the only alternative now left 
them is the additional expenditures for keeping up preventive 
measures already begun on a small scale. 

Eventual drainage of the area adjacent to the railroad for its 
entire length will be necessary before this means of transporta- 
tion can be freed from the present menace to the inhabitants 
of San José. : 

The surrounding towns and barrios must be permanently 
isolated from communication with San José so long as mosquito- 
breeding conditions in them remain as they are now and such 
a large percentage of the population are carriers of the malaria 
germ. 

Oiling and removal of alge from breeding places of Myzomyia 
are at best a mere makeshift until drainage operations can be 
instituted; but one or the other of these processes must be 
constantly employed so long as fresh or salt water remains 
stagnant or even semistagnant in the neighborhood of San José. 

My experience at San José in connection with the breeding 


——————— | 


IX, B, 2 Sanitary Survey in Mindoro 167 


places of Myzomyia rossii and the prevalence of malaria confirms 
the experiences which I have had at Olongapo,* Cervantes, Bon- 
toc, Subig, Iloilo, Cavite, and Baguio. While other species of 
anopheline mosquitoes occur in these localities in very limited 
numbers, M. rossii is extremely abundant in all of them. It is 
the species known to breed in both fresh and salt water,’ and 
it is the only species in abundance in the Philippine Islands, 
which is a proved carrier of malaria here as well as elsewhere.*® 


6. THE DISEASE INDEX. (A) LABORATORY EXAMINATIONS 


By ERNEST LINWOOD WALKER, ARISTON M. GUZMAN, 
and ISABELO CONCEPCION 


In consideration of the part played by malaria in the mor- 
bidity at the San José Estate, it was thought advisable to devote 
chief attention in the laboratory during this investigation to the 
examination of blood with special reference to malarial parasites. 
The examination of feces to determine the incidence of infection 
with intestinal parasites was also undertaken, but owing to the 
limited time at our disposal this series of examinations was 
small. 

Blood smears were made from 1,095 out of the total population 
of about 3,200 persons at San José. One large smear on a 
1- by 3-inch microscopical slide was made from each person. 
Time permitted the thorough examination of only a few of 
these smears in Mindoro, and the remainder were brought to 
Manila for examination. 

The blood smears were fixed in methyl] alcohol, stained with 
Giemsa’s stain, and examined with a 1/12-inch oil-immersion 
objective. With the aid of a mechanical stage, the preparations 
were subjected to a very thorough examination for malarial 
parasites. Attention was directed also in these examinations 
to the presence of other blood parasites, such as spirochetes, 
Leishmania, and filarial larve, and to any abnormalities in 
the blood cells, such as leucocytes containing malarial pigment 
and changes in the red blood corpuscles resulting from anzmia. 
Time did not permit making red and white blood-corpuscle 
counts, but the hemoglobin was estimated in most of the cases by 
the Tallquist scale, and differential leucocyte counts were made 
of a certain proportion of the blood smears which showed 
malarial parasites. In these counts, owing to their large num- 


* This Journal, Sec. B (1907), 2, 513 et seg. 

"Ibid. (1908), 3, 385 et seq. 

° Theobald, Monograph of the Culicide. London (1907), 4, 47; (1910), 
5, 19 (bibliographical). 


168 The Philippine Journal of Science 1914 


ber, only 100 leucocytes were counted. Therefore, the results 
can be considered only as roughly approximate. The white cor- 
puscles were classified in these counts as polymorphonuclear 
neutrophiles, polymorphonuclear eosinophiles, large mononu- 
clears, and lymphocytes. Among the large mononuclears were 
included the transitionals and myelocytes. 

In Table II are summarized the results of the examinations of 
blood smears from 1,095 persons for malarial parasites. In 
this table these persons are classified as well men, well women, 
well children, and fever patients. By “well” is meant persons 
who were up and about, performing their regular duties. 


TABLE II.—The results of the examination of the blood of 1,095 persons for 
malarial parasites. 


| wel | wen | wen | Fever | 


} men, women. | children. | patients. 
= ka *: see fo Fal} ea ae i | 

Number examined -_____-____________ Be Cee 978 | 63 | 23 31 
Positive: } 

Total £42226. 5) steps tees eee sce ee 333 14 10 16 

Plasmiodiwm via 3. = 32 eee ma | 185 7 5 10 

Plasmodium prxcozx (faleiparum) _._..-.-- 189 9 | 7 ll 

Plasmodvum malaris —--=.--=-=-5=22--- 26-5 9 0} 0 0 

Double ‘infection..¢2-- =. 4". 2... ASS 45 2 2 6 

| oD ethene ean no een ai a ee a ee 645 49 13 15 

Percentage of infected persons_______________. $4.05 22.22 43. 48 51.61 


Malarial parasites were found in 373 or 34.06 per cent of the 
1,095 persons examined. If we exclude the 31 persons who were 
ill in bed with fever, the percentage of apparently well persons 
who were harboring malarial parasites in their blood is not 
materially altered, being 33.55 per cent. Since only one blood 
examination was made of these persons and since malarial para- 
sites are not always present in the peripheral circulation in 
sufficient numbers to be seen microscopically, it is probable that 
the true percentage rate is higher than 33.55. 

This investigation was carried on in the dry season. The lack 
of rain together with the efforts of the sanitary officer in charge 
had reduced the mosquitoes within the boundaries of the estate 
toaminimum. With the advent of the rainy season, it is doubt- 
ful whether this condition could be maintained. Consequently, 
the incidence of infection at the time of these examinations was 
probably at its minimum. 

The parasite rate obtained from the examinations of the blood 
of varying numbers of individuals has been reported from differ- 
ent regions, but in most cases this rate has been for endemic 


IX, B, 2 Sanitary Survey in Mindoro 169 


malaria. The conditions in the Panama Canal Zone are, perhaps, 
the most nearly comparable with those in the San José Estate. 
In both places the population consists largely of imported 
laborers who are more or less nonimmune, and in both places 
prophylactic measures against malaria are practiced. Darling 
reported in 1910 (3) that he found malarial parasites in the 
blood of 13 per cent of 276 persons who were up and performing 
their regular duties. Our findings at San José show a much 
higher percentage of infection. 

Considering them by groups, the individuals sick with fever 
naturally show the largest percentage of infected persons, it 
being 51.61 per cent. Well children, as is to be expected, come 
next with a percentage of 43.48 of infection. The well men 
follow with 34.05 per cent of infections. Lastly come the well 
women who were found infected to the extent of 22.22 per cent. 

Of the total 428 infections with malarial parasites, 157 or 
36.68 per cent were tertian, 216 or 50.46 per cent were sub- 
tertian, and 9 or 2.1 per cent were quartan. Double infections, 
chiefly with tertian and subtertian parasites, were found in 50 
persons, and triple infections, consisting of double tertian and 
single subtertian infections, are recorded in 1 man and in 1 child. 

No spirochetes or Leishmania were found in any of the blood 
smears, although a few of the persons examined were East 
Indians. Spirochzetes have been found previously in the blood 
of an East Indian employee suffering from black-water fever 
on this estate.* In a blood smear from one malarial case, a 
single filarial larva was found. 

Gametes of the malarial parasite were found in only 98 or 
26.27 per cent of the 373 infected persons. With few exceptions 
the gametes, when found, were few. Darling (4) has estimated 
that persons whose blood contains more than 12 gametes per 
cubic millimeter, or 1 gamete to 500 leucocytes, are capable of 
infecting mosquitoes, and consequently must be regarded as 
“malaria carriers.” The blood of these 98 persons, and probably 
others, contained more than 12 gametes per cubic millimeter, and 
was, therefore, dangerous to other persons in the presence of the 
proper mosquito host. 

With reference to the red blood corpuscles, the low hemoglobin 
value and the absence of well-marked regenerative changes are 
noteworthy. 

In 266 of the cases of malarial infection in which the hemo- 
globin value was determined, it was as follows: 


* Ashburn, Vedder, and Gentry, Bull. Manila Med. Soc. (1912), 4, 198. 
124316—3 


170 _ The Philippine Journal of Science 1914 


TABLE III.—The hemoglobin index. 


Hzmoglo- 
bin per- Cases. epee 
centage. F 


40 to 49 2 0.75 
50 to 59 18 6. 76 
60 to 69 88 33.08 | 


70 to 79 96 36. 09 
80 to 89 46 17.25 
90 to 100 16 6.01 


Bates in 1913 and previously others have shown that in mala- 
ria the loss of hemoglobin value corresponds very closely to 
the reduction of red blood corpuscles. Therefore, the low hemo- 
globin index indicates a considerable deficiency in red blood 
corpuscles in most of these persons. However, in view of the 
fact that the majority of the persons in whose blood malarial 
parasites were not found also had a low hemoglobin index, it 
seems probable that the anzemia in these people was not due 
wholly to malarial infection, but in part to their general poor 
physical condition. 

In only 102 or 27.34 per cent of the 873 cases of malarial 
infection were anemic or regenerative changes in the red cor- 
puscles recorded. These changes consisted of polychroma- 
tophile corpuscles, basophile punctate corpuscles, and macrocytes. 
In most cases these were not numerous, and in no case were 
nucleated red cells observed. The changes in the red blood 
corpuscles do not appear to be commensurate with the degree 
of anemia indicated by the hemoglobin value in many of these 
persons. 

The results of differential leucocyte counts in 165 of these 
cases of malarial infection are summarized in Table IV. 


TABLE I1V.—Summary of the leucocyte counts. 


Cases falling into different percentage groups. 
Leucocytes. 


0-5. | 6-10. | 11-15. | 16-20. | 21-26. | 26-30. | 31-35. | 36-40. 


INCHLYOD DUG 250-5 - eo ons ee | ee eee 
OsinGphilesa= case a2eee ee eee 64 $1 


10 
1 
Mononuclears and transitionals_-_._-_- 44 53 34 17 5 1 2 
Ibymphocytes: 264s ee - ees a Sei 1 4 23 | 30 2 | 35 18 12 


1X, B, 2 Sanitary Survey in Mindoro 171 


TABLE I1V.—Summary of the leucocyte counts—Continued. 


| Cases falling into different percentage groups. 


Leucocytes. |= eae aaa | SN AE GRRL AE ST | LL SL Ra ak | 
41-45. | 46-50. | 51-55. | 56-60. | 61-65. | 66-70. | 71-75. | 76-80. 


Neutrophiles’—: <2 - +o en Gs ask oe 15 21 26 24 17 16 13 5 


Hosinophiles oats. ss oe ee ee ee oe 2 
Mononuclears and transitionals--------|-------|-------|-------|------- 
TG VIMDNOCYLER Deena ee eeemen ne ———— 7 2 


The variation from the normal in these leucocyte counts is 
best brought out by another table which shows for each type of 
leucocyte the number and percentages of cases having a normal 
count and those having counts below and above the normal. 


TABLE V.—Numbers and percentages of normal and abnormal leucocyte 


counts. 
| Cases having a Cc havi Cases having a 
count below ases Having a! count above 
normal. normal count. normal. 
Leucocytes. pee a oS poe Oe AEs ee 
Num-| Per | Num-| Per | Num-| Per 
ber. cent. ber. cent. ber. cent. 
Neutrophiles (normal count, 60-75 per cent) ___- 107 | 64.84 52 | 81.51 6| 3.638 
Eosinophiles (normal count, 2-4 per cent) _-___-_ 21} 12.72 82) 19.39 112 | 67.87 
Large mononuclear and transitional (normal 
eotiner4-8 per Cent)) .. =o - oa aac acne ne 20] 12.12 61 | 36.96 84 | 50.90 
Lymphocytes (normal count, 20-30 per cent) -___ 49 | 29.69 72 | 43.68 44 | 26.66 | 


From Table V it is seen that in 64.84 per cent of these cases 
of malarial infection there was a diminution in the proportion 
of polymorphonuclear neutrophile leucocytes. This diminution 
in the neutrophiles appears to be more or less compensated for 
by an increase in the eosinophiles and large mononuclear leuco- 
cytes, 67.87 per cent of the cases showing an increase in the 
proportion of the eosinophiles and 50.90 per cent an increase 
in the proportion of the large mononuclears. The lymphocytes 
have a count below and above the normal in about an equal 
number of cases. 

In only 15 of the 373 cases of malarial infection were large 
mononuclear leucocytes containing malarial pigment observed. 

The conclusion of practical importance to be drawn from the 
blood examinations in these cases is that the blood picture, apart 


2 The Philippine Journal of Science 1914 


from the presence of malarial parasites, was of little aid in diag- 
nosing malaria under the conditions existing in this investigation. 
Only in the minority of cases in which there was a large increase 
of the large mononuclear leucocytes and in the extremely few 
cases in which leucocytes containing malarial pigment were 
found was the blood picture significant. 

Thomson (5) has shown that there exists a considerable fluc- 
tuation in the leucocyte counts made at different stages of 
malarial infection. These fluctuations fnclude a decrease in the 
total leucocytes in the peripheral blood during active malaria, 
varying more or less inversely with the temperature; an inverse 
relation between the number of mononuclear leucocytes and the 
temperature; and transient periodic leucocytosis, chiefly of the 
polymorphonuclear leucocytes, in latent malaria. These fluctua- 
tions would probably explain some of the variable differential 
leucocyte counts obtained in these cases. 

The increase in the proportion of the eosinophile leucocytes 
found in a large number of these cases was probably due for 
the most part to intestinal parasites with which these persons 
were infected. 

That so many apparently well adults show malaria parasites 
in their blood in the dry season when mosquitoes have been re- 
duced to a minimum indicates a serious malarial problem for 
the promoters of the San José Estate; and the facts that the 
population of this estate is an imported one and that it is esti- 
mated that 90 per cent of the individuals remain for less than six 
months at San José and then return to their homes in various 
parts of the Archipelago constitute a grave public-health problem 
for the whole Philippines. This estate may be considered as a 
center of infection to which nonimmune persons are constantly 
coming and from which a constant stream of infected persons 
are going out to all parts of the Philippines. Since the mosquito 
that is incriminated in the transmission of malaria in these 
Islands is of wide distribution, these emigrating cases must 
serve as foci of infection in various parts of the Philippine 
Islands. 


THE MICROSCOPIC EXAMINATION OF FACES FOR INTESTINAL 
PARASITES 


The 58 stool examinations made were all of adult males. The 
results of these examinations are shown in Table VI. 


IX, B, 2 Sanitary Survey in Mindoro 173 


TABLE VI.—Examination of feces for intestinal parasites. 


Examination and infections. | Number. |Per cent. || Examination and infections. | Number. |Per Band 
Persons examined ----_---- BS hp sok sae Person infected with— 
Persons infected ---------- 53 91.37 Onyuris ss. ee Sane 1 V2: 
Persons infected with— Entameba-.------- ene 18 22.41 
Hookworms-.----------- 28 48, 27 Balantidium ---------- 2 8.44 
Tt CHuriae sn ea 26 44, 82 AUN OU) ae ete 2 8.44 
VA SCORIB Rae eee aa 21 36. 20 Trichomonas __-------- i 1.72 
Strongyloides-_-------. cil ieee Total infections_____- 105 | 179.31 
| Dibothriocephalus ____- 2 3.44 


The results of this limited number of examinations show a 
percentage of infections with intestinal parasites not varying 
materially from that obtained by other investigators in the Phil- 
ippine Islands. The only special points of interest in these 
examinations are the relatively high percentages of infections 
with Strongyloides stercoralis, Dibothriocephalus latus, and Ba- 
lantidium coli, but the number of persons examined is too small 
for one to lay any great stress on the results. 


LITERATURE 


(1) ASHBURN, P. M., VEDDER, E. B., and GENTRY, E. R. A spirillum in 
the blood of a case of black-water fever. Bull. Manila Med. Soc. 
(1912), 4, 198. 

(2) Bates, J.P. A review of a clinical study of malarial fever in Panama. 
III. Malarial anemia. Journ. Trop. Med. & Hyg. (1913), 16, 
209-213. 

(3) Daruinec, S. T. Studies in relation to malaria. Gov. Printing Office, 
Washington (1910). 

(4) IpEM. Factors in the transmission and prevention of malaria in the 
Panama Canal Zone. Ann. Trop. Med. & Parasit. (1910), 4, 179-223. 

(5) THomson, D. The leucocytes in malarial fever: A method of diagnos- 
ing malaria long after it is apparently cured. Ibid. (1911), 5, 83-102. 


6. THE DISEASE INDEX. (B) CLINICAL EXAMINATIONS 
By W. E. Muscrave, A VAzQuEz, B. GUTIERREZ, and SetH L. Cox 


Of the total population of about 3,200 employees, together 
with women and children, of the San José Estate, 1,110 persons 
were examined clinically. The examinations were more or less 
superficial in character, but sufficient time was taken to deter- 
mine, with a fair degree of accuracy, any gross pathological 


174 The Philippine Journal of Science 1914 


changes that might be present. The persons examined con- 
sisted of: 


Males 1,035 
Females 75 
Total 1,110 


Tabulated by nationalities, they are as follows: 


Nationality. Males. | Females. 
American! =. 2-62 25s-cee eee 17 4 
Bilipinos 2273325 ee 961 67 
Japanese 22 ose tenets 39 4 
Indian: | - ooo sec2 sot 8 0 
Spanish). oes eee ee ee 3 0 
American Negro ____-___-- 1 0 
Mnglishin. 5.2.3 Le 2 0 
Scoteby.2. .¢2 cae pet. ee 1 0 
German. 2c. ae eee 1 0 
SWISS Gc akon ese 1 0 
Groek: 2.6.2 ae Eee 1 0 

Total 23.4. se ee ete 1,085 15 


It must be understood that the part of Mindoro under con- 
sideration formerly was practically uninhabited and that the 
population of 3,200 people represents emigrants from practically 
the entire Philippine Archipelago, so that the examination of 
these people, except in those diseases definitely contracted after 
arrival at Mindoro, represents, to a certain degree, an index of 
conditions found throughout the country. 

The principal points in the physical examination of this entire 
group of persons is indicated below. 


MALARIA 


By reason of incidence and general morbidity and mortality 
effect upon the population in general, malaria ranks third in the 
sanitary problems in the Philippine Islands, infant mortality 
and tuberculosis holding the first and second places, respectively. 

The distribution of malaria is very general. However, there 
are places in the Islands where the disease is unknown, and 
there are a number of other places where the infection is very 
severe, is constantly present, and where the disease has existed 
as far back as we have any record of the country. 

Of all the infected centers, the west coast of Mindoro is popu- 
larly considered to be the worst. As is shown in other places in 


IX, B, 2 Sanitary Survey in Mindoro 175 


this report, the barrio of Mangarin shows a “spleen index” among 
children of 98 per cent, and it is altogether probable that 100 
per cent of the population of this place is infected with malaria. 
The inhabitants of San José, including the officials and em- 
ployees of the Mindoro companies, are made up of a transient 
population recruited from all parts of the Philippine Archi- 
pelago. The incidence of malaria among these people is exceed- 
ingly high, and the morbidity factor is a most important one. 
In the examination of 1,110 employees and other inhabitants of 
this estate, positive enlargement of the spleen was found to be 
present in 26.21 per cent. 

Examination of blood from 1,095 of these employees showed 
malarial parasites in 373, or 34.6 per cent. For a consideration 
of the relative importance of the “‘spleen index” and examination 
of blood smears in the diagnosis of malaria, see the special report 
by Doctors Walker and Cox, page 181. 

The relation of the “spleen index” to ages of patients is 
shown in Table VII. 


TABLE VII.—Relation of spleen index to age. 


Spleen. | 


Cases 
Age. exam- Palpable. Markedly enlarged. 
ined. 
Number. | Per cent. ; Number. | Per cent. 
Hier ivear ss, Seiad ols bea 2 21 3| 14.80 4} 19.00 
UT UD CGS ee Oe Se eee ee ee eee 10 2 20. 00 5 50. 00 
MELO OU CALS Onn ee ne newton acne en aise soe 494 62 12.50 88 17.80 
CLL UT GU G2 pee ee ee ee 393 53 13.50 44 11.19 
SOD UGA ie ee ee a ee ee a er 119 | 13 10. 90 8 6.70 
LUD ae ee ee 48 3 6.25 4 8.33 
Henn venga te gee ee ee! 14 0 | 0 i| 72 
Vt DIR TES ot Seas eas Se os Eee Ae Eee a 2 0 0 | 0 / 0 
Baers Sihrcertas ye Nis Saeed oe yee N 1 1} 100 | 0 0 


It is, of course, probable that a certain small percentage of the 
enlarged spleens are due to causes other than malaria, but even 
eliminating these we still have a malaria incidence that is so 
high as to require radical measures for its reduction. In look- 
ing into the causes of these conditions, we have three important 
sources of infection; first, outside sources. A study of the 
records of people examined shows that they come from all pro- 
vinces of the Philippine Islands, and the incidence of spleen 
enlargement, by districts and countries, is shown in Table VIII. 


176 The Philippine Journal of Science 1914 


TABLE VIII.—Incidence of spleen enlargement, by districts and countries. 


Total Eee Porm Total | Pos- | po 

| District. eee patos cone District. havea! ae cent- 

amined.| index. ee. amined.| index.| 22°: 

Albay £5-- 255 2.4282 33 i 0 0 Malay States____-__- eeRe 1 0 0 

Ambos Camarines ---- 4! 100.0 || Marinduque_____»__--_- 2 1| 50.0 

An tiques-seeee saa ees 47 4 Su5iN Mindanao sean 4 1} 25.0 

Bataan ees ee ees 2 40507} 0Mindoro: oo a 74 36 | 48.6 

Bataness peo 1 0 OU Waal as fi ov 5 fares 1 0 0 

Batangas 2ch5 262-8 19 7 87.0 || Nueva Ecija _.____.--__ 26 7 
Bulacan_____-_-------- 29 13| 44.9 || Nueva Vizcaya ________ 1 1 
Bohol) 24222. 22. Sees 3 4 1 25.0 || Occidental Negros -____- 32 7 
Caniz).-+ asi Se 112 36 32.1 |, Oriental Negros _______ 2 1 
Cavite a2 432.5-8-23° 21 9 42;.b'|| Palawan = sss = 2S 29 0 
Geboiccss22 ees sees 61 16 31.3 || Pampanga _._-.-_-__--- 37 6 
Ceylon tne eae 1 0 0 Pangasinan ____________ 38 14 
Corregidor ___-_------- 4 0 0 Rizalli+s $5252 22 eee 62 11 
nglanid) e220 oso 1 0 0 Romblonts=-2-2 8225-55-38 5 1 
Guan sons eee 1 0 0 Samar oe se eee 4 1 
PMonGlwlas es ec 1 0 0 Simgapore._......_..._- 3 2 
HocosiSar 2.2 ae ea 15 4 26.6 || Sorsogon ..-<- 5-2 24255 6 1 
llocos Norte-__-_-------- 1 0 0 Surigao n= 1 0 
Hoile/s2c2-- ee 221 38 QT Zi) Deblaa SS ase 8 2222 8 5 3 
Indias 432 se) 3 0 0 dd Eh er eee ete 153 49 
DADA ooo ae 38 10 2950) Wayabas 322 ek 8 ul 
Wagun® 2-25. eee 6 8 50.0 || United States__________ 7 0 
ia Unions. 1 0 0 Zamboanga __---------- 1 0 
tS 1 nr mi ea 9 4 | 1 BONO NOG @iVeR cee! = el 2 0 


These marked variations in the percentage of infection of 
persons coming from different provinces indicate that a certain 
number of these people already were infected with malaria 
before their arrival in Mindoro. This statement is inferen- 
tially corroborated by subsequent statistics made by “preém- 
ployment” examination of people who are going to Mindoro. 
These laborers are recruited from various provinces of the 
Islands, and, roughly speaking, the “‘spleen index” among this 
class of people is averaging about 10 per cent. Therefore, it 
seems logical to conclude that approximately 10 per cent of the 
malaria of Mindoro is imported through the new recruits con- 
stantly arriving from all parts of the Islands. 

The second source of infection is found in the barrios near 
San José. From the extensive infection of these places and 
the close travel association between them and San José, it is 
believed that approximately 5 per cent of the San José malaria 
may be accounted for in this way. Thirdly, the remaining 15 to 
20 per cent of the total infected with malaria at San José are 
accounted for by the spread of the infection in San José and 


—_——_— 


IX, B, 2 Sanitary Survey in Mindoro 77 


the outlying camps of the corporations. The remedies for this 
malarial situation will be found in another part of this report. 


TUBERCULOSIS 


The work of the Philippine Islands Antituberculosis Society 
has revealed a rather startling condition of affairs regarding 
the incidence of tuberculosis among Filipinos. Most of the 
work of this society has been done in the city of Manila, and, 
consequently, deals largely with patients resident in Luzon; very 
little is known, statistically, of the incidence of this disease in 
the provinces. 

Physical examination of 1,110 people, included in this report 
with reference to tuberculosis, gives the following result: 


Positive 331 
Doubtful 10 
Negative 768 
Not recorded 1 

Total 1,110 


In as much as these patients come from a large number of 
provinces as shown in a previous table, the figures quoted in 
the above statistics show a markedly high incidence and a re- 
markable general prevalence of tuberculosis. As the average 
residence of these people in Mindoro is, approximately, three 
months, it is assumed that the majority of them contracted this 
disease before they emigrated to this island. The enormous 
prevalence of tuberculosis is a very important factor both in 
the economic and in the health problem of Mindoro. With the 
present barrack system of quartering the employees in San José 
and the overcrowded condition of most of these barracks, such 
an enormous percentage of tuberculosis infection means disaster 
unless the situation is relieved in some manner. It is a well- 
known fact to those who have been studying the tuberculosis 
problem of this country for many years that the average laborer 
will not acknowledge that he is ill and will continue his ordinary 
vocation while suffering from this disease until hemorrhage, 
excessive fever, or general weakness makes it impossible for 
him longer to make an effort to perform his work. The 
tuberculosis situation may very readily be controlled by a careful 
preémployment examination of applicants for positions at Min- 
doro, excluding all those who have definite signs of the disease, 
and the condition of those already on the ground may be markedly 
improved by the establishment of a different residence system 


178 The Philippine Journal of Science 1914 


from the one now in use. It is altogether probable that tuber- 
culosis is a more important factor in the decreased capacity for 
labor among employees at San José than is malaria or any other 
one physical abnormality of the people. 


HEART AND BLOOD VESSELS 


Physical examination of the circulatory system of the 1,110 
people gives the following result: 


Valvular disease: 


Positive 78 
Doubtful 4 
Negative 1,028 
Total 1,110 
Hemic murmur 20 
Chronic myocarditis : 4 
Tachycardia Lol 


There are no available statistics of the incidence of cardiac 
diseases among supposedly healthy people of the laboring classes 
in other countries, and it is, therefore, impossible to say whether 
the rather striking picture shown in these statistics is an unusual 
one. 

The question of etiology and incidence of diseases of the heart 
and blood vessels in the tropics has received practically no con- 
sideration in medical literature. These conditions are among 
the important considerations in the medical work in this country. 
Whatever the difference in incidence between valvular diseases 
in the tropics and in the temperate climates may be, it seems 
very likely that the etiological factors are somewhat different. 
Statistical work dealing with this subject, based upon findings 
in our clinics of the College of Medicine and Surgery and the 
Philippine General Hospital, now are being made, and sufficient 
work has already been done to justify the statement that there 
exist in the tropics destructive valvular diseases of the heart that 
are not explained by the usual etiological consideration in these 
diseases. At this time, only two of the causative agents will 
be referred to; namely, environment and malnutrition. 

The rather constant low blood pressure found among natives 
of the tropics as well as among foreign residents in the tropics 
is, in all probability, environmental in origin. Whatever is the 
exact cause, there is in the tropics a relaxed condition of mus- 
cular tone, with dilated peripheral blood vessels, slight tendency 
to cedema in dependent parts of the body, and a sluggishness of 
gastrointestinal metabolism and absorption as well as in the 


: 


: 


1X, B, 2 ; Sanitary Survey in Mindoro 179 


function of the internal organs of secretion. This condition 
of affairs is intimately associated with myocardial changes, and 
it is altogether probable, although not an experimentally sub- 
stantiated fact, that toxic conditions result from this general 
stasis which have positive effect in myocarditis in addition to 
the mechanical effects on the valves as a result of some cardiac 
dilatation. 

Applicants for employment who have valvular diseases of the 
heart or show circulatory incompetency in any form should 
be excluded and not allowed to emigrate to Mindoro. 


BERIBERI 


Examination of 1,110 persons with reference to the incidence 
of beriberi gives the following result: 


Positive 3 
Doubtful 16 
Negative 1,091 

Total 1,110 


In addition to the findings in this tabulation, we are creditably 
informed that at no time in the history of this estate has beriberi 
assumed an important rdle among the disease conditions. This 
is rather striking in view of the modern conception of the rela- 
tionship of polished rice to this disease. 

The development of Mindoro being in a very elementary stage, 
the local supply of fish and other foodstuffs is mostly imported 
from Manila and adjacent islands. 

The diet of the people of the San José Estate consists almost 
entirely of polished rice, fish, and a few vegetables and condi- 
ments. In other words, the diet is about as poor as that of any 
class of Filipinos in the Islands. 


HOOKWORMS 


Stool examinations were made in only 58 cases, and 28 or 
48.27 per cent of these were found infected. Most of these 
infections were light in character, and clinical inefficiency in 
these patients was but little, if at all, lower than the general 
average. We feel confident that hookworm infection at the 
present time is a small factor in the clinical inefficiency of em- 
ployees of the San José Estate. 


CLINICAL INEFFICIENCY 


The general physiological efficiency of the laborers and other 
inhabitants of San José is very low, and it is believed that 50 


180 The Philippine Journal of Science 1914 


per cent of the efficiency as compared with the normal ability 
of an average healthy Filipino would be a conservative estimate 
of conditions in this respect found in Mindoro. There are 
several reasons for this low state of nutrition and development 
in addition to that part of it which is explained by the high 
incidence of malaria, tuberculosis, and other diseases. 

The principal factors concerned in the production of this in- 
efficiency are the habits and customs of the people, particularly 
the poor quality and small amount of food consumed by the 
average Filipino of the working classes. The low efficiency 
stated is based on the general appearance of the people, their 
early exhaustion and inability to perform a reasonable amount 
of manual labor, and upon certain physical findings and a series 
of hemoglobin estimates. The relation of the percentage of 
hemoglobin estimates to enlarged spleens is shown in Table IX. 


TABLE 1X.—Relation of the percentage of hemoglobin estimates to 
enlarged spleen. 


—— 


Hemoglobin 
estimates. 

Cases of 
enlarged 

Per cent] spleen. 

Persons. jof haemo- 
globin. 

91 100 13 

81 90 47 

71 80 78 

61 70 | 81 

51 60 82 

| 41 50 6 

i 81 | 40 2 

Not recorded_-_-_-_- 32 

291 


Examination of this table indicates that malaria as determined 


by enlarged spleen is not the principal factor in the reduction of | 


hemoglobin percentage, neither is this true with regard to tuber- 
culosis. The condition is more far reaching and has to do, as 
stated above, with the poor nutrition and under development of 
the people as a whole. 

Intimately associated with the lowered vascular tone mentioned 
above, either as cause or as effect, is the high incidence in mal- 
nutrition; and the results of this malnutrition, obviously, are 
both mechanical and toxic. The causes of malnutrition are two. 
First, an economic condition which makes it impossible for 
people of the working classes properly to nourish themselves and 


Ow 


IX, B, 2 Sanitary Survey in Mindoro 181 


their families; and, secondly, the inability properly to assimilate 
satisfactory food products, provided these are obtainable. 

The general question of clinical inefficiency is largely an eco- 
nomic one, but nevertheless it is one of the most important 
phases of the complex problem of health and sanitation in Min- 
doro, as well as in other parts of the Philippine Islands. 


7. A COMPARISON OF THE SPLEEN INDEX WITH THE MICRO- 
SCOPIC EXAMINATION OF THE BLOOD FOR MALARIAL PARASITES 
IN 1,003 PERSONS 


By ERNEST LINWOOD WALKER and SETH L. Cox 


There are two methods in general use for obtaining the index 
of malarial infection in any region. One of these is by micro- 
scopic examination of the blood of a certain portion of the 
inhabitants for malarial parasites; the other is by the determi- 
nation by palpation of the number of the individuals having 
enlarged spleens. Both methods have their application and their 
limitations. The microscopic examination of the blood is espe- 
cially applicable for determining the incidence among nonim- 
mune persons. The advantages of this method are that it is 
more apt to discover recent infections and that its results are 
direct and. unequivocal; its disadvantages and limitations are, 
first, the time required to make an adequate research of the blood 
of a sufficient number of individuals; and, secondly, the fact 
that the parasites cannot always be found in the peripheral 
blood, especially in chronic cases. The spleen index is especially 
applicable to the determination of the endemicity in a malarial 
region. It is a convenient and quick method; but it is subject 
to the errors that enlargement of the spleen in the tropics may 
be due to causes other than malaria, that it is considered reliable 
only when applied to children between the ages of from 1 to 
10 years, and that it may not indicate recent infections in which 
the spleen had not yet become enlarged. 

In the laboratory examinations of about one-third of the 
population of San José, a relatively large proportion of indi- 
viduals was found to harbor malarial parasites in the blood, and 
in the physical examinations of these same persons a nearly 
equal proportion was found to have enlarged spleens. There- 
fore, it has seemed worth while to compare the two series 
in order to determine how far they coincided. This could 
readily be done, as the two series of examinations were made 
on the same persons and the individuals of each series could be 
identified by their company number. 


182 The Philippine Journal of Science 1914 


Certain peculiar conditions must be taken into consideration, 
however, in comparing the spleen rate and the parasite rate of 
the inhabitants of San José. 

The population of San José consists chiefly of adult males and 
of relatively few women and children. Consequently, both the 
blood examinations and the spleen palpations in these series 
have been made chiefly of adult males. Neither the spleen rate 
nor the parasite rate is considered reliable in diagnosing en- 
demic malaria unless applied to children between the ages of 1 
and 10 years. 

Moreover, it is to be noted that the population of San José 
is for the most part not a permanent but a shifting one. The 
laborers of this estate are practically all imported, both from 
all parts of the Philippine Islands and from abroad. They are 
chiefly Filipinos, but include some Japanese and East Indians. 
A few Americans and Europeans are employed to superintend 
the work of the laborers and to perform the skilled work on 
the plantation. The Filipino laborers, for one reason or another, 
usually remain only for a short time at San José. After work- 
ing for a few months, they return to their homes and are 
replaced by freshly imported men. A certain proportion of 
them return from time to time to be reémployed on the estate. 
The company estimates that 90 per cent of their laborers remains 
in their employ less than six months, that 9 per cent remains 
more than six but less than twelve months, and that 1 per cent 
remains more than one year. 

This circumstance modifies the interpretation to be placed on 
the results of these examinations. The population being con- 
stantly shifting, the new arrivals, unless coming from a malari- 
ous region, would be nonimmunes. Therefore, the blood and 
spleen examinations of the inhabitants of San José would show 
not the endemic index of that region, but the incidence of infec- 
tion with malaria among the more or less nonimmune adults. 

Of the 1,064 persons, exclusive of those who were ill with 
fever, of whom microscopic examination of the blood was made, 
357, or 33.55 per cent, showed malarial parasites; and of the 
1,110 persons whose spleens were palpated 291, or 27.11 per 
cent, showed enlargement. Therefore, if the two series should 
coincide, approximately equal results would be obtained by the 
two methods of diagnosis. 

Considering first the total number of persons of whom both 
spleen palpation and microscopic examination of the blood were 
made, Table X shows the spleen rate, the parasite rate, and the 
percentage of agreement between the two series. 


1X, B, 2 Sanitary Survey in Mindoro 183 


TABLE X.—The relation of splenic enlargement to malarial parasites in the 


blood. 
ohare Boren: 
alaria’ age o 
Spleen. nach ea. Cases. eres 
ment. 
Palnablen 26 sere ree erste ne eet, ema ene Ma ew a Pee Fe + 39 } 
42.29 
ID Fey oR A ee ee Si See aN eee et ae oe ees re a 0 53 
Muchienlarcedesssee ean Da Ee ae + 66 } 
40. 24 
DED cp eye Re De HO Ih a ee 0 98 y 
otalteniar@ed eee ee eee ce We eo ok + 105 } 
1.01 
TC) eee ee eee a SRO as Oe ie oki 2 SR 0 161 ; 
INF CUM eet |e Ee ee I pee + 224 | 
01 
LION coyote lt ai ee ore ee ne a ee a er 0 523 iW 


From Table X it appears that the agreement between spleen 
rate and the parasite rate is not very close. The percentage of 
agreement is greatest in those cases that showed no splenic 
enlargement, being 70.01 per cent; it is markedly less in those 
cases that show a splenic enlargement, being 41.01 per cent; 
and it is slightly less in those cases which have a much enlarged 
spleen than in those where the spleen is only palpable, being 42.24 
and 42.39, respectively, in the two series. This relationship 
between spleen index and microscopic findings may be expressed 
in another way. Of 256 persons who had enlarged spleens 
only 105, or 41 per cent, showed malarial parasites in their 
blood; and of 329 persons who showed malarial parasites in 
their blood only 105, or 31.91 per cent, had enlarged spleens. 

It is true that 50 of the persons who had enlarged spleens 
but in whose blood malarial parasites were not found gave a 
history of attacks of malaria within a year, and it is probable 
that others, who gave no history of malaria, had suffered from 
previous attacks of malaria. It is further probable that 
some of the persons who were negative microscopically for 
malaria were nevertheless infected. Taking these facts into 
consideration, the percentage of agreement between those who 
had enlarged spleens and those who were or had previously been 
infected with malaria would be increased. 

Since both the spleen rate and the parasite rate are generally 
considered to be more trustworthy when applied to children 
between the ages of 1 and 10 years, it is of interest to compare 
the two methods of diagnosis in these patients grouped accord- 
ing to ages. For reasons previously stated, the majority of 
these people were adult males; therefore, the number of children 
between the ages of 1 and 10 years included in these examina- 
tions is small. 


184 The Philippine Journal of Science 1914 


TABLE XI.—Relation of splenic enlargement to the presence of malarial 
parasites in the blood in persons grouped according to age. 


Parasites. Spleen. 
Fescent> 
Age. age o: 
Much agree- 
Infec- Palpa- |Not en- 
. Cases.| en- ment. 
| tion. fareedt ble. |larged. 
Cases. | Cases. | Cases. 
Ab tov 10) Semen ee eet SAI Ra { 3k 11 a) 1 8| 72.70 
0 16 3 Pe ll 68. 75 
11 to. O0ivears on ee tee ee | os 163 40 17 106 34.96 
0 275 46 18 211 76. 72 
21 te SOlyenes aie 2c: Saw ea | ete 113 17 18 78 30. 97 
0 254 35 23 196 77.16 
31:t640 years. co ee ee ee | i 21 2 2 17 19. 04 
0 a 9 7 66 70. 48 
41''to) 60) years oS. o 4 Fo ee eee a 1" |--------|-------- 12 |---------- 
0 29 3 2 24 82.75 
BL tov60 years: - oe ect ae a ae ee ee 3 5 |--------|-------- 5 |---------- 
Ole Oiundes oe | 9} 81.81 
GLt0) TiyeGaras 225.5) ee ee eee + 0 |--------]-------- eae! Deere 
0 | eR (teen ee 2) 100.00 
+ Ui hs i ok | a O |-----. --|--------]---~ ----]---------- 
0 Woe a Bit epee ee 100. 00 
Agé unknowns. 22. i> 2 eS eee ee { at 4 |-------- 1 3 20. 00 
| 0 4 loo sqoe| eases 4} 100.00 


From Table XI it appears that the percentage of agreement 
between parasite rate and spleen rate in those persons who 
showed malarial parasites in the blood is greatest between the 
ages of 1 and 10 years and constantly decreases in the older 
age groups; while, on the other hand, the percentage of agree- 
ment in those persons who were negative for malarial parasites 
is lowest in the age group of from 1 to 10 years and progressively 
increases in the older age groups. Nevertheless, the percentage 
of total agreement between the spleen rate and the parasite rate 
is greater in children than in adults. 

Therefore, the results of this comparison of the spleen index 
and microscopic examination of the blood for malarial parasites 
indicate that, under the conditions of this investigation, the 
microscopic examination of the blood is the more reliable method 
of determining the incidence of infection, first, because more 
cases were discovered by this method; and, secondly, because 
the microscopic diagnoses of the positive cases are unequivocal. 
These conclusions are applied only to the determination of the 
incidence of malarial infection at San José and not to the endemic 
index. 


= wae = 


IX, B, 2 Sanitary Survey in Mindoro 185 


8. SUMMARY AND CONCLUSIONS 


The following extract from the preliminary report of the 
commission is a brief summary of the health conditions of the 
west coast of Mindoro, with the important recommendations 
for their improvement. 


STATEMENT OF CONDITIONS 


The sanitary problems of the Mindoro properties are a part 
of, and inseparable from, similar problems of the municipality 
of Pandorocan and the whole west coast of Mindoro. The 
municipality has a population of some 5,500 people, of which 
about 3,200 are employees or otherwise inhabitants of the camps 
of the corporation located at San José. 

Both the sanitary and the social and economic problems 
must be solved by experimentation, because pioneer work 
is required under conditions which have no precedent. To give 
an idea of the magnitude of these problems, it is only necessary 
to state that they are equal to, or greater than, those encountered 
in the construction of the Panama Canal. 

The San José interests have an area of between 210 and 260 
square kilometers of territory, which is bounded on the east 
by the mountains of Mindoro, the inhabitants of which are 
wild men who appear to be very extensively infected with malaria. 
The property is continuous along the coast line with a number 
of the most insanitary barrios, whose inhabitants are very 
extensively infected with malaria, tuberculosis, hookworms, and 
other tropical diseases. In the Canal Zone, with a slightly larger 
geographical area than the San José Estate, all efforts are 
centered on the digging of the canal, while in San José the 
naturally more complex problem is amplified, because of efforts 
to cultivate the land with irrigation. Finally, very unwise 
administration in the early part of the development work of this 
location has added materially to the amount of infection, partic- 
ularly malaria, which must be eradicated before satisfactory 
health conditions can be established. 

The studies of the commission may be briefly summarized as 
follows: 

The incidence of malaria among the inhabitants of San José 
is approximately 35 per cent of the total population, and this 
infection is kept up by recurrence in old malaria cases and by 
constant introduction of new cases from without and from 
outlying camps and by new infections among the inhabitants 
of San José. 

124316——4 


186 The Philippine Journal of Science 1914 


The incidence of tuberculosis is about 30 per cent; hookworm 
infection from 45 to 50 per cent; clinical anemia about 90 per 
cent; with a large share of other less important diseases and 
conditions. 

Most important of all, the physiological efficiency of the em- 
ployees and the inhabitants of San José may be comparatively 
estimated at 50 per cent, using as a basis for this calculation 
the normal efficiency of the average healthy Filipino. 

The social and economic conditions of the inhabitants of San 
José are not as satisfactory as they should be. The opinion is 
advanced that the barrack system of housing employees and 
their families is a mistake in the tropics among tropical races, 
and it undoubtedly is a mistake in the absence of careful and 
exacting police and sanitary protection. The construction of 
these barracks is of a fairly satisfactory character, but under 
present conditions they are dirty, badly kept, and overcrowded, 
and show a complete absence of any attempt in the establishment 
of home life among the inhabitants. 

The management of the kitchens and mess halls is not con- 
sistent with elementary sanitary requirements. For example, 
one kitchen within 60 meters of the hospital contains myriads 
of flies; it is dirty; and it is used as a storeroom for food and for 
the soiled clothing of the cooks and assistants and as a bath- 
room by muchachos and children. Conditions in the mess hall 
proper are but very little better. 

The caloric value of the foodstuffs eaten by the majority of 
the employees, whether in the mess halls or in their own houses, 
is far below the minimum physiological requirement consistent 
with manual labor. This condition, as is well known, is general 
among the lower classes of Filipinos. It is one of the as yet 
unsolved problems of this country. 

The medical department, as a whole, is doing splendid work, 
particularly in its mosquito-suppression work. However, with 
the enormous amount of malarial infection now prevalent in the 
camp, the methods of work now in use will not begin to meet 
the situation during the rainy season with the consequent in- 
crease in the number of mosquitoes. The entomologic survey 
shows that the inhabited portion of the properties is very well 
policed regarding mosquitoes, but on all sides just without this 
zone are innumerable breeding places filled with anopheline 
mosquitoes. With the advent of the rainy season, it will be 
impossible to maintain the present mosquito-free zone without 
the expenditure of an unreasonable sum of money. 


IX, B, 2 Sanitary Survey in Mindoro 187 


9. RECOMMENDATIONS 


Successfully to cope with the sanitary problem connected with 
the Mindoro properties demands a definite organization of a 
sanitary department with its personnel, equipment, and sup- 
plies; the delegation of a definite authority; and sufficient ap- 
propriation to carry on the work. The head of this department, 
whether he be an employee of the company or of the Government 
or of both, unquestionably should have the authority and perform 
the duties of a district health officer, not only for the Mindoro 
properties but for all outlying barrios. This department should 
direct its efforts along the following lines: 


1. The prevention of reinfection by the importation of malaria-free laborers 
’ and by restricting the intercourse between the inhabitants of the 

protected zone and infected persons in near-by territory. 

2. Efficient hospitalization for sick people. 

3. Satisfactory free dispensary service for those who are not ill enough 
to require hospital care. 

4. A visiting service for the location of carriers and other infected persons. 

5. The systematic treatment of all persons (compulsory if necessary) harbor- 
ing the malarial parasite. 

6. Constant application of approved methods for the reduction of anopheline 
mosquitoes. 

7. A sanitary corps whose duties should be the cleaning up of new camps 
before such camps are occupied by laborers. 

8. Supervision of the housing and feeding of employees, their families, and 
all other inhabitants of the zone. 

9. Carefully planned and systematic reports from all divisions of this 
department. 


As is well known, the propagation of malaria depends upon the 
integrity of a three-link chain, and the breaking of any link 
or the reduction in the strength of all three links embodies 
the whole principle in combating this infection. The three links 
are: 


1. The infected person. 
2. The proper variety of anopheline mosquitoes. 
8. The nonimmune person. 


The campaign against this infection, therefore, must be along 
three principal lines and along all of them at the same time: 


1. The suppression, as far as possible, of the propagation of anopheline 
mosquitoes. 

2. The protection, as far as possible, of nonimmune persons from mosquito 
bites, particularly at night. 

3. Most important of all, the prevention of the introduction of new cases 
from without and the thorough treatment of all infected cases whether 
new or recurrent, within the inhabited zone, together with the protection 


188 The Philippine Journal of Science 1914 


of these persons during their period of infection against the bites of 
malaria-carrying mosquitoes. 


Every case of malaria that applies for treatment and every 
case that can be found in quarters or in the field should be treated 
actively and aggressively until the patient no longer is a carrier 
of the malarial parasite. The character of this treatment is a 
detail subject to discussion, but the main point is that it should, 
and if success is to be attained must, be carried out as indicated. 

For the prevention of the introduction of malaria from without, 
two important things are necessary: 


1. There should be arrangements for proper physical examination of all 
laborers at their points of embarkation and before their services are 
accepted by. the company. 

2. Communication with the badly infected barrios adjacent to the pe ee 
must cease, and in the enforcement of this phase of the question 
generous Government support will be necessary. As will be indicated 
in the final report of the commission, a number of these barrios should 
be transferred to more healthful locations, and aggressive and per- 
sistent municipal effort should be employed in cleaning up the other 
infected places. 


SANITARY DEPARTMENT 


The following outline of a sanitary department was recom- 
mended by the commission and, in part, has been placed in 
operation by the directorate of the Mindoro properties: 

Chief physician.—Directly responsible to the manager. To be 
in general charge of all sanitary matters, including the admin- 
istration of hospital, dispensary, mosquito suppression, general 
camp sanitation, and supervision over water supply, food supply, 
etc. All employees of the sanitary department to be subject to 
the chief physician. 

Superintendent.—Directly responsible to the chief physician. 
His duties should be as follows: Administrator—in charge of 
records, property, employees, cooks, commissary, etc. 

There should be one bookkeeper, two clerks, one cook, and as 
many muchachos as the work calls for. 

Chief nurse.—Directly responsible to the chief physician. 
To be in charge of housekeeping, hospital, dispensary, nurses, 
nurses’ home, employees’ quarters, kitchen, commissary, and 
linen. Two female and two male nurses would be required 
on the staff. 

Resident physician.—Directly responsible to the chief phy- 
sician. His duties to consist in professional work of hospital 
and dispensary. 

Visiting physician.—Directly responsible to the chief phy- 


1X, B, 2 Sanitary Survey in Mindoro 189 


sician. To visit officials and families, employees sick in quarters, 
and all sick persons. In charge of mosquito brigade and sani- 
tary corps. 

Chief sanitary inspector.—Directly responsible to the visiting 
physician. To have charge of the carrying out of all rules 
relating to general sanitation. Five sanitary inspectors should 
assist him in the performance of his duties. 

Because the work of the sanitary department is so compre- 
hensive and the work to be done is of such magnitude and so 
urgent, it is recommended that the chief physician organize 
his department into four individual divisions, namely: (a) Ad- 
ministration division; (b) hospital and dispensary ; (c) mosquito- 
suppression division; and (d) sanitary division. 

Administration division.—This should be in immediate charge 
of the superintendent of the hospital, and should consist in the 
institution and maintenance of a satisfactory system of records, 
professional as well as those of accountability of property, etc.; 
these records, of course, to include the preparation of the neces- 
sary pay roll, requisitions, receipts, etc. connected with the run- 
ning of the department. It should have charge of the general 
questions of employees, housing, property, supplies, commissa- 
ries, mess hall, cooks, kitchens, and dormitories for attendants. 


. The superintendent must, of course, have the necessary book- 


keeper, clerks, and other employees for the proper administration 
of the division. 

Hospital and dispensary.—This division should be directly 
under the resident physician for his professional work, the 
chief nurse for her professional work, and the superintendent 
for the administrative work—all these three, in turn, being 
responsible to the chief physician. The division of the sanitary 
department needs some strengthening in personnel, as is in- 
dicated above, and its duties should be increased successfully to 
care for all sick people of the community. It is not possible to 
conduct this division with any degree of efficiency under the 
present construction and arrangement of the hospital. A rough 
sketch outline of suggested additions to the hospital has been 
furnished. The committee is fully aware of the fact that ad- 
ditional construction is expensive, but the most important con- 
dition to be met in Mindoro at the present time is to get rid 
of as much of the malaria now there as is possible. To do this 
will require expansion of the hospital, the establishment of a 
fairly large free dispensary service, and good codperative work 
on the part of the sanitary department. With increased hospital 
space and increased personnel, the hospital days of the average 


190 The Philippine Journal of Science 1914 


case of malaria should be reduced from two to four days, the 
patients, of course, being required to come to the free dispen- 
sary daily, every other day, or twice a week, as the case may be, 
for subsequent treatment until the disease has been completely 
eradicated. 

Mosquito-suppression division.—This division should be im- 
mediately under the direction of the visiting physician who, in 
turn, is responsible to the chief physician. 

Mosquito-suppression measures of economy and of approved 
value should be carried out by the chief mosquito inspector 
and such additional employees as are necessary. Modification and 
method of procedure in this division as well as in all others 
should be indicated by written circulars of instruction from the 
chief of the department. Such detailed rules and regulations 
as will insure daily inspection of breeding places of mosquitoes, 
location of infected mosquitoes, infected patients, etc. should 
be made very explicit and be very rigidly enforced. This 
division of the work in San José is already in a very satisfactory 
condition. The chief mosquito inspector might well be a male 
trained nurse who has had special instruction in this branch 
of work. 

The mosquito-suppression division and the sanitary division 
should be the advance guard in opening any new camp or new 
spot of civilization or in any other method of extension of the 
activities of the company. 

The sanitary division.—This division is a very important one. 
The chief sanitary inspector should be a male trained nurse 
who has had special instruction in general sanitary work. He 
should be directly responsible to the visiting physician and have 
such additional help as may be necessary. He should inspect 
daily every habitation and every camp and report on every 
case of illness, particularly persons suffering with fever and 
requiring hospital or free dispensary attendance. He should 
see that sleeping quarters are not overcrowded, are kept with 
at least a semblance of cleanliness, and that they are properly 


ventilated. He should enforce rules regarding the distribution © 


and use of water; the collection and disposal of sewage and 
garbage; should locate and destroy the breeding places of flies; 
and should carefully inspect and control the sanitary condition 
particularly of kitchens, mess halls, and other places where food 
refuse of any kind is apt to accumulate. 

It hardly seems necessary to indicate further the numerous 
minor details connected with the efficient administration of these 
two divisions. 


4 
: 
\ 
‘ 


it 


IX, B, 2 Sanitary Survey in Mindoro 191 


The above outlined organization will give a unit that should 


’ be sufficient to meet the requirements of the situation, provided 


the necessary governmental coéperation can be secured in con- 
trolling the external problem. Recommendations covering this 
phase of the subject are that: 


1. The municipal government of Pandorocan be moved to Santa Teresa. 

2. The barrio of Mangarin be transferred to Santa Teresa. 

8. The chief physician of the San José Estate be given the official designa- 
tion and authority of a district health officer. 

4, On account of the increasing incidence of malaria fever in various parts 
of the Philippine Islands, the Director of Health be requested to 
declare malaria a dangerous communicable disease within the meaning 
of the law, in order that the usual methods of eradication may be 
enforced when necessary. 

5. The Government make arrangements, either by conferring authority upon 
the officers of the San José Estate or otherwise, to provide satisfactory 
police and legal protection for the inhabitants of Pandorocan. 


10. APPENDIX 


The following series of questions submitted in letters addressed 
to the attorney and chief physician, respectively, of the Mindoro 
properties, together with the answers to these questions, are 
so important in understanding the social, sanitary, and economic 
problems of Mindoro that they are given in full: 


1. What was the average population during the years 1911 and 1912; 
separate figures for men, women, and children? 

The Mindoro Company: No census was kept during the years 1911 and 
1912. 

San José Estate: 1911, men about 2,000. Very few women and children 
here this year; 1912, men about 2,200, women about ‘200, and children 150. 


2. The average number of laborers on your pay rolls during the above 
period? 

The Mindoro Company: 628 per month. 

San José Estate: 1911, about 500; 1912, about 2,000. 


38. The average number of days worked per month by these employees? 
The Mindoro Company: Sixteen days. 
San José Estate: 1911, about fifteen days; 1912, about seventeen. 


4. Average weekly income per person based upon the total adult male 
population? 

The Mindoro Company: 5.14 pesos. 

San José Estate: Population too variable to give this as asked, but the 
average weekly income for the worker in 1911 was 3.85 pesos and in 1912 
4.50 pesos. 


192 The Philippine Journal of Science 1914 


5. Average daily wage for unskilled labor; average daily wage for 
skilled labor; what percentage of employees are classed as skilled labor? 

The Mindoro Company: The average daily amount actually earned per 
man on roll was 73.5 centavos. About one-third of all employees on rolls 
were skilled workmen. 

The San José Estate: In 1911 the labor was practically unskilled and 
the average daily wage was 51 centavos. In 1912 we had about 10 per 
cent skilled labor, the average wage for which was about 1.80 pesos; for 
unskilled labor it is about 60 centavos per day. 


6. What percentage of your employees remains in your employ for less 
than six months? What percentage for more than six months and less 
than one year? What percentage remains more than one year? 

The Mindoro Company: No reliable data available. 

San José Estate: 90 per cent; 9 per cent; 1 per cent. 


7. What was the total number of new employees engaged during the 
years 1911 and 1912, by years? 
' The Mindoro Company: No reliable data available. 

The San José Estate: Approximately 1,800 each year. 


8. What was the total number of employees leaving your service during 
the same period of time? 

The Mindoro Company: No reliable data available. 

San José Estate: About 125 a month. 


9. What percentage of the wages of unskilled labor is paid back to 
the company: (a) For food, (6) for rent, (c) for water, (d) for light, (e) 
for all other purposes? 

The Mindoro Company: No reliable data available. During the years 
1911 and 1912 the Mindoro Company operated a store which catered to 
the employees of the San José Estate, Mindoro Company, and Manila 
Construction Company. The last company employed some 300 men in 
construction work on the sugar mill from July to December, 1911, who were 
not carried on the estate or company rolls. The store not only handled 
foodstuff, but clothing and household necessities; therefore, any attempt 
to give figures on money paid back for the specific commodities would be 
idle guess work. No charge was made for water or houses. 

San José Estate: (a) about 25 per cent; (b) none; (c) none; (d) about 
3 per cent; (e) none. 


10. Approximately what percentage of the wages paid unskilled labor do 
you estimate as being paid to private tiendas? 

The Mindoro Company: It is estimated that considerably less than 10 
per cent was paid to private tiendas. 

San José Estate: About 10 per cent. 


11. Based upon the sales through the company’s stores and agents, what 
would you consider the average daily money value of subsistence per capita 
for the total population, including men, women, and children? 

The Mindoro Company: Owing to the absence of census figures and as 
no store record along these lines was kept, it is impossible even to approxi- 
mate daily money value of subsistence per capita of total population. 

San José Estate: For adults about 25 centavos per day. 


a 


IX, B, 2 Sanitary Survey in Mindoro 198 


12. Is the quality of rice (red and white rice) purchased optional 
with inhabitants, and approximately what percentage of the quality of 
this staple article of food is used and what is the average price of each? 

The Mindoro Company: Nothing but white rice was sold in store at an 
average price of 35 centavos per ganta. 

San José Estate: Optional. The prices are graded according to prices 
ruling the rice market in Manila. 


13. What is the average price at which fish is sold? 

The Mindoro Company: 35 centavos per kilogram. 

San José Estate: 40 centavos a kilogram in the store. In the private 
tiendas, when fish is scarce, the natives, unless very closely watched, will 
run the price up to any figure they can get. 


14. Is the supply of green vegetables kept as complete as consistent with 
the market, and how do the prices compare, for these articles, with prices 
for similar articles in Manila and other parts of the Islands? 

The Mindoro Company: Green vegetables were handled when possible 
and sold at practically Manila prices. 

San José Estate: Yes, about the same as throughout the provinces. 


15. What class of fuel is used by the majority of your unskilled labor; 
what is the source and what price is paid? 

The Mindoro Company: The fuel used consists mainly of scrap lumber 
for which no charge is made by the company. 

San José Estate: Coal and wood free, picked up about the place. 


16. What is the average expense for each unskilled laborer up to the 
time he begins work? ‘ 

The Mindoro Company: Each laborer costs the Company approximately 
10 pesos to land on premises. 

San José Estate: About 13 pesos. 


17. What precautions are taken to avoid the introduction of the various 
contagious diseases with the importation of new laborers? 

The Mindoro Company: No medical examination was made of new 
laborers up to the first of the year. 

San José Estate: None. 


18. What are your age-limit requirements, minor and major? 

The Mindoro Company: No age limit was observed up to the first of 
the year. 

San José Estate: None. 


19. Approximately how much was spent for sanitary improvement 
including personnel, equipment, supplies, mosquito brigade, etc. during the 
year 1911 and during the year 1912, and what is your monthly expense 
at the present time? 

The Mindoro Company: Approximately 40,000 pesos were expended in 
sanitary work during 1911 and 1912, exclusive of the hospital equipment 
and maintenance. About 2,000 pesos are expended per month at the present 
time for sanitary work, exclusive of hospital expenses. 

San José Estate: From the beginning of the San José Estate, April, 
1911, to December of the same year, over 7,000 pesos were expended. For 
the year 1912, over 65,000 pesos were expended. At the present time the 
average monthly expense is 3,500 pesos. 


194 The Philippine Journal of Science 1914 


20. Please itemize your present monthly sanitary expenses under the 
following headings with such additions as you deem advisable: (a) Salaries 
and wages; (6b) equipment and supplies; (c) construction; (d) medicine, 
dressing, etc.; (e) subsistence; (f) miscellaneous. 

In this connection, I should like to have a list of your personnel connected 
with the sanitary department, with salaries and wages of all employees 
except those of unskilled labor. 

Mindoro Company: The sanitary and hospital work is at present per- 
formed by the San José Estate force. 

San José Estate: (a) Salaries and wages about 3,500 pesos. (6) Equip- 
ment and supplies about 150 pesos. (c) Construction referred to the 
Mindoro Company under whose auspices this work is carried on. (d) Med- 
icines, dressing, etc. It is not possible to answer this question as these 
goods are bought in large quantities and it is difficult to say how much 
is used per month. (e) Subsistence about 450 pesos per month. (f) Mis- 
cellaneous expenses amount to 38 pesos per month. Appended is a list of 
the personnel with their salaries: Physician etc., 850 pesos; nurses, 300 
pesos; attendants, 363 pesos; sanitarians, mosquito chasers, etc., 623 pesos; 
miscellaneous, 120 pesos. The difference between these figures and the 
3,500 pesos quoted above is paid to the police, the workers on the incinerator, 
and the daily labor employed in cleaning camp and performing various 
duties of a like nature. 


21. What charges, if any, are made against employees for medical and 
hospital attention: (a) For skilled labor, (b) for unskilled labor? 

The Mindoro Company: No charges are made against workmen for hos- 
pital and medical attendance. 

San José Estate: No charges are made to either skilled or unskilled 
labor, and time goes on during accident. 


22. Is the time lost on account of illness a charge against the employee 
or against the company, and, in the latter event, during what period of 
time does the company continue to pay wages when the employees are ill? 

The Mindoro Company: All laborers are on a daily basis, and are paid 
for actual work performed. 

San José Estate: Time lost from accidents in the company’s service is 
paid by the company. 


23. What charges, if any, are made for professional and hospital 
attendance of the families of the employees: (a) Of skilled labor, (6) of 
unskilled labor? 

The Mindoro Company: No charge is made for hospital cr medical 
attendance in the cases of family of employee. 

San José Estate: None. 


24. Is medical attendance for employees reported absent on account of 
illness compulsory, and what methods for investigating such reported illness 
are in practice? 

The Mindoro Company: Medical pendants for employees absent on 
account of reported illness is not compulsory. 

San José Estate: Yes. The camps are visited daily by police and 
sanitarians, and all cases are reported to the medical authorities and 
treated either at home or in hospital as the judgment of the doctor in 
charge directs. 


og 


IX, B, 2 Sanitary Survey in Mindoro 195 


25. What methods are employed for the supervision of sleeping and 
residence accommodations of employees (with particular reference to the 
prevention of overcrowding) ? 

The Mindoro Company: This service is at present performed by the 
San José Estate. 

San José Estate: The same course of instruction as outlined in answer 
to question 24, but, in spite of all precautions, the natives will overcrowd 
and it is almost impossible to stop them from doing so. 


26. What regulations are in force regarding the importation and sale 
of (a) alcoholic beverages, (b) patent medicines? 

The Mindoro Company: No regulations are in force regulating im- 
portation of alcoholic drinks or patent medicines. Beer and native wines 
are sold at the San José store only. 

San José Estate: (a) Open market. (b) Not kept on the place. 


27. What methods, if any, are employed to improve sanitary conditions 
along educational lines? Is there any school system available for the 
children of the community? 

The Mindoro Company: See answer to question 20. 

San José Estate: None. School building but no teacher. 


28. What is the nature of available facilities for religious worship? 
Mindoro Company: There are no facilities for religious worship. 
San José Estate: The padre of Caguray makes periodical visits. 


29. What are the available forms and customs regarding recreation 
and amusement? 

The Mindoro Company: The company has a baseball diamond, tennis 
court, and moving picture show available to all. 

San José Estate: Cockpit and dance hall for the natives. 


30. What requirements and provisions regarding the collection and the 
disposal of sewage and garbage, and what methods are employed to 
enforce the regulations? 

The Mindoro Company: See answer to question 20. 

San José Estate: Garbage cans are placed at each house, and the contents 
are emptied daily by carts and burned. Violation of any sanitary law 
is punished by heavy fine. 


31. In your opinion, what are the causes for the high morbidity rate 
of the employees of your corporation? 

The Mindoro Company: Referred to Mr. George H. Fairchild. 

San José Estate: As it is very probable that the diseases responsible 
for the high rate of morbidity here were in a large measure brought to 
this place, it seems to me that an inquiry into the sanitary condition of 
the places where these natives come from would best answer this question. 


32. What recommendations, if any, have you to suggest to the com- 
mission that if embodied in this report would be of assistance to you in 
improving sanitary conditions? 

The Mindoro Company: Referred to Mr. George H. Fairchild. 

San José Estate: Remove the surrounding sources of infection, such 
as Mangarin. 


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ILLUSTRATIONS 


PLATE I 


Mangarin and San José, Mindoro, showing areas found to be infested by 

Myzomyia rossii Giles. Crosses in red indicate actual breeding place of 

Myzomyia rossi. Area inclosed in solid red line indicates region practi- 

cally free from breeding places of this mosquito. Area inclosed between 

_ dotted red lines and coast line is that which from its appearance seemed 

favorable to mosquito breeding. The most dangerous area lies at present 

along the railroad. 

ae TEXT FIGURES 

___-Fig. 1. Mindoro Island, showing the location of San José Estate. 

2. The townsite of San José Estate, Mindoro. 
: 197 


Pes une 


* 


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SANITARY SURVEY IN MINDORO.] [Puin. Journ. Sci., IX, B, No. 2. 


LEGEND 
---{ limits within which Malaria 
Mosquitoes were found. 
Limits of area freed from 
Malaria Mosquitoes. 
+ { Ploceswhere Myzomyia Mosquitoes 
were found breeding in abundance 


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MANGARIN AND SAN JOSE \ 
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PLATE I. Mangarin and San José, Mindoro, showing areas found to be infested by 
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Considered from the viewpoint of prac ') 
3] utility, Mr. Walker's gar’ Industry 
in the Island of Negros is one of the most 
important papers published by the Bureau’ 
of Science, This volume is a real ocontribu- 
tion to the subject; it is not a mere com 
pilationy, for the author was. in the field and 
Smead the conditions of whith he 
writes. 


a 


A MANUAL OF PHILIPPINE SILK 
CULTURE | 


By Cartes S. BANKS 


Order. No. 413. Paper, 53 pages, 20 
plates, $0.75, postpaid, 


In/A. Manual. of Philippine Silk Culture =. 
are presented the results of severe aes , 
actual work with silk-produoing Lox 
gether with a. description of the new Pri 
Dine race. i 


ry 
' 


THE PHILIPPINE 


JOURNAL OF SCIENCE 


B. TROPICAL MEDICINE 
Vou. IX JUNE, 1914 No. 3 


SOME DATA CONCERNING THE MEDICAL GEOGRAPHY OF 
THE PHILIPPINES + 


By FERNANDO CALDERON 


(From the College of Medicine and Surgery, University of the Philippines, 
and the Philippine General Hospital) 


The literature on the medical geography of the Philippines 
is very scant, and any investigator who would search in the 
archives and libraries for treatises concerning the demography 
of the different regions of the Archipelago would perhaps find 
an article here and there about a tropical affection and its 
predominance in certain localities or an essay about the diseases 
most frequently observed in certain regions of the Philippines, but 
never would he find any work which deals with the demography 
and sanitary condition of the Archipelago as a whole nor would 
he find any article on the geographic distribution of the diseases 
that afflict the people of the Philippines in the different regions 
of the Archipelago. The only work that pretends to approach 
the medical geography of the Philippines was written by Antonio 
Codorniu,? a member of the Spanish Medical Corps in these 
Islands, and this work is at the most not more than a tentative 
one on the “description of the Philippine conditions which have 
an influence over the human body under the special circumstances 
of individuality with respect to the inhabitants of the Islands.” 
His more or less authentic observations, however, are worth 
repeating here. He says that— 

1. The northern provinces are more healthful than those situated south 


of Manila, and the Visayan Islands, in spite of their being in a low latitude, 
» are the most healthful place of all. 


*Read before the Congress of Filipino Physicians, held in Manila, 
February, 1914. 
? Medical Topography of the Philippine Islands. Madrid (1857). 
126334 199 


200 The Philippine Journal of Science 1914 


2. Aside from the Province of Misamis, whose salubrity can only be 
determined from doubtful data, Abra is the most healthful and Nueva 
Vizcaya is the most mortiferous of all. 

8. The provinces are classified according to their health status as follows: 
healthful, moderately healthful, and most healthful; and under each class 
are mentioned the different provinces. 


Later on, in 1889, José Solis? and others edited in Manila 
a pamphlet written by the professors of the medical corps, and 
in this work the authors show that the diseases most prevalent 
in the Jolo Archipelago are typhoid fever, dysentery, and malaria. 
Other contributions worthy of mention are by Manuel Rogel 
Lebres * and Pedro Robledo Gonzalez.° The first author posi- 
tively affirmed that in the Visayan provinces alone are found 
10,000 lepers, while the latter asserted that the above figures 
represent the number of lepers in the whole Philippines in 1880, 
and explains the Japanese origin of the disease by the fact 
that in the latter part of the eighteenth century about 100 
Japanese lepers who were banished by the Mikado reached Luzon 
and since their arrival leprosy spread rapidly all over the Islands. 
In the three centuries of Spanish dominion, the above are the 
only available data concerning the medical geography of the 
Philippines. 

In the fifteen years of American régime, numerous and varied 
articles have been published about the matter. 

Washburn ° says: 

In the classification of climates based on the size and extent of masses 
of land, oceanic, insular and continental, the climate of the Philippine Is- 
lands is largely that of the first two cases, oceanic and insular. Nowhere 
is the land distant from the seacoast more than 60 miles. The moderating 
influences of the great bodies of sea water are therefore operative. On 
account of climatic influences, the climate of the Philippines is widely 
different from those of tropical Africa, South America and Asia in the 
same latitude. As a rule, the smaller the island the more equable the 
climate throughout the day and the year. The climate of the greater part 
of the Philippine Archipelago is for this reason comfortable and hygien- - 
ically favorable for the treatment of many diseases. 

In the temperate zone an insular, mild, or equable climate is frequently 
a health resort. 

Observations in the Manila Observatory show that while the temperature 


*Contribucién al estudio estadistico-demografico-higiénico de Jol6. Ma- 
nila (1889). (Contribution to the study of vital statistics and hygiene of 
Jolo.) 

“Leprosy in the Visayan Provinces (1897). 

* Leprosy in the Philippines (1902). 

‘The relation between climate and health with special reference to Amer- 
ican occupation of the Philippine Islands. Read before the second annual 
meeting of the Philippine Islands Medical Association in 1904. 


1x,B,3 Calderon: Medical Geography of the Philippines 201 


as indicated by the thermometer at sea level is practically the same 
throughout the Philippine Archipelago, it is higher in some regions than 
in others during the months of April, May and June which cover the 
periods. There exists a considerable difference in climate between the 
coastal regions of the islands of the Archipelago and the elevated regions 
of Luzon and Mindanao. Mountain climate of these two islands is similar 
to that in Baguio, Luzon, whose altitude is 4,777 feet and whose average 
temperature and humidity are relatively low during the hot months of 
March, April, May, and June owing to the prevailing winds blowing from 
the China sea during that season. The climate of this region is ideal from 
November to June. 


The article of Jerome B. Thomas’ describes the principal 
diseases found in 1903 and 1904 among the Igorots and Ilocanos, 
who live in Baguio and the neighboring mountain regions. 
These diseases are: Beriberi in benign form, bronchitis which is 
most common during the months of January and February, 
influenza, malaria, rare cases of amecebic dysentery, gastro- 
enteritis due to indigestion, some cases of cerebrospinal menin- 
gitis, various kinds of skin diseases, and parasitism. 

In regard to the intestinal parasites, we should mention the 
interesting article of Garrison.2 This investigator examined 
microscopically the feces of 4,106 inmates of Bilibid Prison of 
Manila, and found that 3,447, or 89 per cent, had one or more 
varieties of intestinal parasites. The author suggests that, 
although these prisoners come from all parts of the Islands, it 
would be interesting to make a minute investigation in the dif- 
ferent provinces of the Archipelago and without doubt it could be 
demonstrated that in certain regions there is an excess of intes- 
tinal parasites. In the same year, 1908, Ross S. Rissler and 
Liborio Gomez presented a paper about the campaign they made 
against the intestinal parasites in Las Pifas and other towns of 
Cavite and Rizal; of 6,018 individuals they found that 5,406, or 
89.84 per cent, harbored intestinal parasites. In another paper® 
these authors, having examined the feces of 10,000 individuals, 
arrived at the conclusion that parasitism is a universal affection 


"Notes on diseases encountered in Baguio, Benguet, P. I., and the adja- 
cent highlands of central Luzon, including revised excerpts from the advance 
sheet of a report to the Secretary of the Interior. Read before the Manila 
Medical Society on February 6, 1905. 

* The prevalence and distribution of animal parasites of man in the Phil- 
ippine Islands, with a consideration of their possible influence upon the 
public health. Read before the fifth annual meeting of the Philippine 
Islands Medical Association in 1908. 

* Prevalence of intestinal parasites in Rizal and Cavite and in Cagayan 
Valley. Read at the assembly of the Far Eastern Association of Tropical 
Medicine in Manila in March, 1910. 


202 The Philippine Journal of Science 1914 


in the Philippines. This conclusion may seem too radical, but 
it is confirmed by the fact that the examination of the feces 
of all the patients admitted in the obstetrical department of 
the Philippine General Hospital has shown that about 80 per 
cent of the women have intestinal parasites. 

Two articles that deserve mention are Filariasis and elephan- 
tiasis in southern Luzon’? and Notes on the distribution of 
Filaria nocturna in the Philippine Islands by Phalen and 
Nichols. In the first paper, the authors arrive at the conclu- 
sion that filariasis is not, as is generally believed, an un- 
common affection in the Philippines; that in the Bicol provinces 
exists the largest focus of filariasis known with the exception, 
perhaps, of Davao where there is even a larger focus; and that 
what they reported as Microfilaria bancrofti, generally known 
as Filaria nocturna, is the most common form of filaria in the 
Philippines. In the second paper the authors show that in the 
valley of Kilani River in Albay Province there seems to exist 
an endemic center of filariasis, while the surrounding regions 
are not as highly infected. Probably small foci of infection may 
also be found in Samar, Leyte, and Mindanao. 

Lastly, I shall mention the interesting article by Willets’? in 
which, after describing the topography and the inadequate sani- 
tary conditions that prevail in the Batanes Islands, north of 
Luzon, as well as the habits and customs of the inhabitants, he 
alleges that the diseases most commonly registered in that 
isolated region of the Archipelago are: Tuberculosis; pneumonia; 
pleurisy; skin diseases, especially a chronic ulcer of the thigh; 
rheumatism; Bright’s disease; and malaria, most commonly of 
the pernicious type, which appears usually from July until 
December. 

The above is all I could glean from the literature about the 
medical geography of the Philippines which exists in the archives 
and libraries I had the opportunity to examine. However, due 
to the kindness and spirit of codperation of Dr. Victor G. Heiser, 
the Director of Health, it has been possible for me to present 
here the interesting vital statistics of health as written below: 

The Director of Health states that, among the diseases having 
an important influence upon the mortality of the Philippine 
Islands and which science is able successfully to combat, are the 
following. 


” This Journal, Sec. B (1908), 3, 293. 
“ Tbid. (1908), 3, 305. 
* Ibid. 1913), 8, 49. 


i i a Se te aml las Reet ae 


1x,B,3 Calderon: Medical Geography of the Philippines 203 


BERIBERI 


Beriberi, in round numbers, causes approximately 5,000 deaths 
per annum. There is much evidence available to show that 
beriberi in mothers affects the nutritive value of their milk, and 
this is believed to be one of the causes for taon among children. 
That the mother’s milk is closely associated with the high infant 
mortality in the Philippines is further borne out by the fact that 
three times as many breast-fed infants die as bottle-fed infants, 
which is directly contrary to the experience in Europe and 
America. If it is admitted that taon in the child is caused by beri- 
beri in the mother, this one disease alone is responsible for at 
least 25,000 deaths per annum. There is now much experimental 
and practical evidence that the continued consumption of white 
rice as a staple article of diet is responsible for beriberi. In 
Government institutions in the Philippines, during the time 
that white rice was used as a Staple article of diet, there 
were on an average more than 600 deaths per annum; whereas, 
since unpolished rice has been used, there have been no deaths 
from beriberi in those institutions. Such evidence is further 
strengthened by the fact that at Culion, for instance, after the 
use of polished rice was again begun, beriberi appeared soon 
afterward and disappeared again when unpolished rice was sub- 
stituted for it. In Cebu Island, where corn is the staple article 
of diet, there is practically no beriberi and taon is exceedingly 
rare. In Manila, on the other hand, where the use of polished 
rice is very common, the highest mortality from beriberi is found. 
Here, then, is a disease, a method of avoiding which is ready for 
trial, and only the codperation of the medical profession is 
needed in order to bring the matter to a satisfactory test. 


MALARIAL FEVER 


Malarial fever is another disease which the statistics show 
to be responsible for at least 25,000 deaths per annum. That 
malaria is transmitted only by the bite of a mosquito that has 
previously bitten a person suffering with malarial fever is now 
universally admitted. Furthermore, it is well known that if 
quinine be promptly administered an attack of malarial fever can 
be avoided. Here, then, are two methods by which malaria in 
the Philippines could be combated: First, by eliminating the 
breeding places of mosquitoes; and, secondly, by the prophylactic 
use of quinine in communities in which malaria prevails and by 
the prompt treatment of those who have been infected with this 
disease. Geographicaly, malaria is most common in the follow- 


204 The Philippine Journal of Science 1914 


ing provinces, where the death rate from this disease during 
1912 was as follows: Cagayan, per 1,000 of population, 7 deaths; 
Oriental Negros and Ambos Camarines, each 6; and Pangasinan, 
Laguna, Batangas, and Ilocos Norte, each 5 deaths per 1,000. 
It was least common in Zambales, Surigao, and Samar, each 
of which had 1 death per 1,000 of population during this 
year. Recently, the Bureau of Health, with the approval of the 
Governor-General, has made arrangements for the distribution 
of quinine through municipal treasurers, from whom 0.3 gram 
quinine tablets may be obtained for 1.5 centavos** each. As there 
are over 700 municipal treasurers in the Islands, this means that 
there will be 700 agencies for the more general distribution of 
quinine. 
SMALLPOX 


Smallpox is a disease that in former times was responsible 
for at least 40,000 deaths per annum. Since systematic vaccina- 
tion has been carried on throughout the Islands, the mortality 
has been reduced to a comparatively few hundreds, but the 
necessity for constant vigilance in this disease has been most 
forcibly demonstrated by the events of the past two years. On 
account of the fact that revaccination and the vaccination of 
newborn children were not carried out in some of the municipal- 
ities, an outbreak of smallpox occurred which in one province 
alone caused over 700 deaths. One of the principal difficulties 
in the Philippines is to preserve the ordinary glycerinated vaccine 
virus, and by actual experience it has been found that it is of 
very doubtful value after it has been removed from an ice 
box for a period longer than ten days. Much experimenting has 
been done, and recently a powdered vaccine has been developed 
which it is hoped will keep for a much longer period of time, but 
the danger from infection is greatly increased by the use of.this 
kind of vaccine, so that the glycerinated virus used before a 
week has expired since it left the ice is still the safest. A careful 
review of the statistics of the Philippine Islands shows that 
smallpox infection apparently exists everywhere in the Islands, 
and it will make its appearance in any community in which there 
are unvaccinated persons. 

LEPROSY 


Leprosy is a disease that has been responsible for at least 
1,000 of deaths per annum and probably for the new infection 
of a like number of persons. Practically all of the lepers of 


" One centavo equals 0.005 dollar United States currency. 


1x,B,3 Calderon: Medical Geography of the Philippines 205 


the Philippines have now been segregated at Culion, and new 
cases that appear are transferred to that colony as rapidly as 
practicable. Apparently the number of new cases of this disease 
has already been reduced to less than 700 per annum, and by 
persistently carrying out the policy of isolation there is much 
hope that in time to come this terrible scourge can be entirely 
eliminated from the Philippine Islands; or, at least, its ravages 
reduced to proportions that will compare with the more advanced 
countries of the world. In this connection, it is interesting to 
observe that, while Cebu Island has approximately one-tenth of 
the population of the Philippine Islands, yet it furnished over 
one-half the lepers that have been collected. 


AMCBIC AND BACILLARY DYSENTERIES 


Ameebic and bacillary dysenteries still prevail to a very great 
extent in the Philippines. The latter occurs almost annually in 
epidemic form, and is most prevalent in those provinces in 
‘which there is a poor water supply. For instance, on Catan- 
duanes, a small island with a population of 39,288 and which 
is noted for its poor water, there were several hundred deaths in 
1912 from bacillary dysentery ; whereas, in towns in which there 
is good artesian water, bacillary dysentery is becoming less and 
less common. This one disease alone affords most concrete proof 
of the necessity of improving the water supply of the Philippines, 
and if one active doctor in each community would constantly 
keep this matter before the attention of the residents no doubt 
great improvement could be brought about in the course of a 
few years. Ameebic dysentery up to comparatively recent times 
was believed to be a disease which mostly afflicted Americans 
and Europeans, but the autopsies made at the city morgue showed 
that 33 per cent of the persons afflicted with this disease were 
Filipinos. There is already much evidence accumulating, how- 
ever, to show that améebic dysentery is becoming less frequent. 
This is probably somewhat influenced by the improved hygiene 
which is taking place among the masses, making it a criminal 
offense to use human excrement as a fertilizer or insecticide. 
Owing to the fact that diagnoses can only be verified in a few 
isolated instances in the provinces, there are as yet no reliable 
data available as to the localities in which this disease is most 


mmon. 
cera FILARIA 


Another disease, which as yet has not attracted great attention 
in the Philippine Islands but which is of more and more im- 
portance, is filariasis. For instance, an examination of 2,629 


206 The Philippine Journal of Science - 1914 


prisoners at Bilibid showed that 402 were afflicted with filaria. 
The highest percentage of those affected was among the pris- 
oners from Leyte Province, which had 35 per cent. Sorsogon 
follows with 31 per cent, Bohol with 27 per cent, Albay with 27 
per cent, and Ambos Camarines with 24 per cent. The smallest 
percentage occurred among the prisoners from Pangasinan, this 
being 0.94 per cent; next is Pampanga, with 2 per cent; and 
Bataan with 3 per cent. The factors which influence these in- 
fections are not yet well understood. It is, of course, known 
that the disease is transmitted by mosquitoes, and probably the 
same measures which would succeed in eradicating malarial mos- 
quitoes would greatly reduce the incidence of filariasis. 


TYPHOID FEVER 


Typhoid fever has been styled the disease of modern civiliza- 
tion. In the Philippines, it is becoming of more and more im- 
portance from year to year. Many positive cases, the diagnoses 
of which have been verified by laboratory examination, have 
occurred in the following provinces: Ambos Camarines, Bulacan, 
Cavite, Ilocos Sur, Iloilo, Mountain, Pampanga, Pangasinan, 
Rizal, Tayabas, La Union, and Zambales. The mortality rate 
from diseases like typhoid, ameebic and bacillary dysenteries, 
cholera, and other intestinal diseases clearly points the way to 
the fact that if proper disposal of human excrement could be 
brought about thousands upon thousands of lives could readily 
be saved. 

TUBERCULOSIS 


It is estimated that there are at least 40,000 deaths annually 
from tuberculosis in the Philippine Islands. General educational 
measures have been taken against this disease almost since the 
beginning of American occupation. In 1910 the Philippine 
Islands Antituberculosis Society was organized, and an appro- 
priation was made by the Legislature. The Bureau of Health 
has established hospitals for chronic tuberculosis patients in 
Manila and a hospital in Baguio for incipient cases. The sub- 
ject has been actively taught in the schools, and the newspapers 
have disseminated much useful information, but so far the desired 
result has not been brought about. If the Philippine Islands 
desires to be considered in the rank of progressive countries, a 
campaign much more active than that of the past must be un- 
dertaken. The best methods of combating this disease are now 
the same in all countries, and the success that has been had in 
the United States and Europe in reducing the ravages of this 


1x,B,3 Calderon: Medical Geography of the Philippines 207 


disease should encourage us in the Philippine Islands to resume 
our efforts to bring tuberculosis under control. In a country 
that is blessed with an equable climate like that of the Philip- 
pines, it should be possible by a general educational campaign 
to bring about open-air sleeping, and this alone would no doubt 
prove a great factor in reducing the incidence of this disease. 

This brief review of a few of the preventable diseases in 
the Philippine Islands shows clearly that, if modern knowledge 
which is already available could be successfully applied, a hun- 
dred thousand deaths in the Philippine Islands could be prevented 
annually. The physicians of the Philippines have a wonderful 
opportunity before them. 

These are the data furnished by the Director of Health. But 
to make a more thorough investigation, I personally addressed 
the following questions to a great majority of the physicians 
located in the provinces. 

A. Which are the diseases, whether medical or surgical, that predominate 
in your province? 

B. In your locality which diseases predominate in certain months of the 
year and what are their causes? 

C. Could you indicate the local and general causes of the predominance of 
such disease or diseases in your district? 

D. What treatment could be practiced in your district to correct the prepon- 
derance of such affections? 

Ninety-five physicians from the provinces had the kindness 
to respond to the questions sent to them, and I cannot let this 
occasion pass without expressing my profound gratitude for 
their codperative spirit. 

To synthesize the accumulated data as a result of the above 
questions, I have made a tabulation of the answers received, and 
I here submit to your indulgent consideration the following 
tables. 

In Table I we can see that the three diseases which predom- 
inate in the whole Archipelago are pulmonary tuberculosis, 
malaria, and dysentery. It can be said that in each region, 
each province, each town, and even in each barrio these three 
affections are the most terrible diseases that afflict the Filipino 
race. Next in frequency and predominance are beriberi, the 
intestinal parasites, gastroenteritis, bronchitis, and broncho- 
pneumonia. From Table I we can see, also, that the diseases of 
the different provinces of the Archipelago are identical. 

There is no doubt, however, that a closer study of the medical 
geography of the different provinces would reveal the presence 
of some unknown local and typical affections that characterize 


208 The Philippine Journal of Science 1914 


certain regions of the Archipelago. To prove this assertion, I 
may cite as examples, what I personally observed; namely, in 
Calbayog, Samar Province, the rarest clinical forms of asca- 
riasis; in Ormoc, Leyte, a disease called kolo-kolo which I had 
not observed in any other locality; and in Jaro, Leyte, many 
cases of simple exophthalmic goiter in women who inhabited 
that mountainous district. Furthermore, we have near us in 
the town of Parafiaque, Rizal, the disease locally known as buvas, 
or yaws, which is a common affection of the inhabitants. 

If we look at Table II which condenses the answers to question 
B (which diseases predominate in your locality in certain months 
of the year and what are their causes?), we can readily see that 
as a whole there are no seasonal diseases in the different parts 
of the Archipelago, but that the same pathological entities with 
slight variations prevail during the entire year. The truth of 
this assertion, however, seems to me questionable. Neverthe- 
less, we notice that in Ilocos Sur, Iloilo, Leyte, and Occidental 
Negros, and perhaps in the rest of the Archipelago, smallpox, 
measles, varicella, and all forms of gastrointestinal affections 
predominate during the hot months—February, March, April, 
and May—while cases of grippe and bronchitis are more numer- 
ous during the months of November, December, and January— 
the coldest months of the year. 

The answers to questions C and D are not included in this 
paper, as they contribute no additional information on the causes 
and remedies of the known diseases in the Philippine Islands. 

All agree in admitting that the wide distribution of pulmonary 
tuberculosis in the Philippines is due to the ignorance of the 
masses in questions of hygiene and that such ignorance is 
responsible for the overcrowding of the churches, the giving of 
banquets in houses where tuberculous persons have just died, 
the abuse of alcoholic drinks, the lack of personal cleanliness, 
the habit of sleeping with closed windows, the lack of care in 
isolating tuberculous patients, as well as that the inevitable 
sequele of poverty is the insufficient nutrition of the body. With 
these observations, it is clear that in combating this terrible 
disease there should necessarily be an energetic antituberculosis 
campaign along educational lines. 

It is believed by the majority of thinkers that the wide distri- 
bution of malaria is due to the abundance of mosquitoes in pools 
and in other stagnant waters found all over the Philippines and 
the best remedy for this disease consists in drainage, the drying 


1x,B,3 Calderon: Medical Geography of the Philippines 209 


of all marshy places, the education of the people in the use 
of the mosquito net, and the gratuitous distribution of quinine. 

Dysentery in its bacillary and amcebic forms is the necessary 
consequence of the imbibition of the contaminated waters con- 
sumed by the majority of our people, and this affection may be 
prevented by the construction of artesian wells. 

Beriberi and all forms of gastroenteritis so common in our 
children may be diminished by the establishment of milk stations 
in the provinces, and the wide spread of skin diseases among 
our masses may be controlled by personal cleanliness and 
hygienic modes of life. 

The above is a brief summary of the diseases which on account 
of their geographical importance occupy a prominent place in 
the pathology of the diseases found in these Islands. 

I fully appreciate the difficulty of the task of making a medical 
geography of any region. In civilized countries more advanced 
than ours, every enterprise of this nature has met with serious 
difficulties and many obstacles; but these difficulties are even 
greater in our country where the data of vital health statistics 
are deficient and inaccurate, as you may have observed in the 
tables which show that provinces have not more than two or 
three diseases. The reason for this is not because these prov- 
inces are more healthful than others, but because there is a lack 
of data in the answers received to the questions distributed all 
over the Islands. As long as we allow 80 per cent of our munic- 

-ipalities to exist without qualified physicians to direct the sanita- 

tion of the towns where the illnesses of the inhabitants are not 
properly diagnosed and as long as we do not succeed in estab- 
lishing an efficient public-health service whose beneficial in- 
fluence may extend all over the Archipelago, we can never 
expect the present conditions of affairs to improve, for the 
diagnosis in the death certificates is made by the municipal 
secretary or in his absence by the municipal police, and such 
certificates form the material available to anyone who would 
undertake the difficult task of writing a medical geography of 
the Philippines. 

Let us have faith, however, in the wisdom and patriotism 
of our legislators, and let us hope that this condition of affairs 
will not last forever, but that before long we shall have a 
reorganization of the present sanitary system by which, by 
means of a correct knowledge of the true medical geography 
of the Philippines, we may perceive the effective diminution 
of the mortality in these Islands. 


210 The Philippine Journal of Science 1914 


TABLE I.—Geographical distribution of diseases in the Philippine Islands. 
[Very incomplete records from the reports by municipal physicians.] 


Albay: Bronchitis; cystitis; convulsion, infantile; dysentery; eclampsia; 
erysipelas; gastroenteritis; gonorrhea; herpes; leucoderma; malaria; 
metritis; parasites, intestinal; rheumatism; scabies; scrofula; syphi- 
lis; tuberculosis, pulmonary. 

Batangas: Beriberi, infantile; bronchitis; dengue; dysentery; gastroenter- 
itis; malaria; parasites, intestinal; tetanus, umbilical; tuberculosis, 
pulmonary. 

Bohol: Abscesses; arthritis, tubercular; asthma; blennorrhagia; bronchitis; 
chlorosis; cloriza; convulsion, infantile; cystitis; dysentery, amcebic; 
epilepsy; furunculosis; grippe; hemorrhoids; malaria; rheumatism; 
scabies; tuberculosis, pulmonary; thrush; urethritis. 

Bulacan: Abscesses; anthrax; beriberi, acute and chronic; beriberi, infan- 
tile; bronchitis; convulsion, infantile; dysentery; erysipelas; furuncu- 
losis; gastroenteritis in children; herpes; infections, gastrointestinal; 
malaria; phlegmon; scabies; tuberculosis, pulmonary. 

Capiz: Abscesses; bronchitis; dengue; dysentery; furunculosis; malaria; 
metritis; rheumatism; tuberculosis, glandular; tuberculosis, pulmonary. 

Cavite: Beriberi; colitis; convulsion, infantile; dysentery; gastroenteritis 
in children; malaria; rickets; tuberculosis, pulmonary. 

Cebu: Abscesses; anemia; beriberi; dermatosis; dysentery; eclampsia; en- 
teritis, acute; gastroenteritis; grippe; leprosy; malaria; measles; men- 
ingitis, simple; rheumatism; smallpox; syphilis; tetanus; tuberculosis; 
typhoid. 

Ilocos Sur: Anemia; anthrax; bronchitis; furunculosis; hemorrhoids; in- 
fections, gastrointestinal; malaria; measles; parasites, intestinal; rheu- 
matism; tuberculosis, pulmonary; varicella. 

Iloilo: Abscesses; dysentery; eclampsia; fevers, gastrointestinal; furuncu- 
losis; gastroenteritis in children; grippe; harelip; hemorrhoids; hernia; 
malaria; measles; parasites, intestinal; Pott’s disease; tonsillitis; tuber- 
culosis, pulmonary. 

Isabela: Malaria. 

Jolo: Dysentery; malaria. 

Laguna: Abscesses; beriberi; bronchitis; convulsion, infantile; cysts; dys- 
entery; eczema; gastroenteritis; gastrointestinal disorders; pleurisy; 
tuberculosis. 

La Union: Gastroenteritis; malaria; skin diseases; parasites, intestinal; 
tuberculosis. 

Leyte: Beriberi; bronchitis; convulsion, infantile; dysentery; enteritis; 
typhoid; gastrointestinal disorders; malaria; measles; parasites, in- 
testinal; tuberculosis, pulmonary; ulcers. 

Mindanao: Abscesses; appendicitis; autointoxications; beriberi, adult; beri- 
beri, infantile; dengue; dysentery, amcebic; enteritis; gastroenteritis; 
gonorrhea; grippe; hydrocele; malaria; parasites, intestinal; phleg- 
mon; rheumatism; skin diseases; syphilis; tabes dorsalis; tuberculosis, 
pulmonary; typhoid; yaws. 

Mindoro: Abscesses; bronchitis; dysentery; hemorrhoids; hydrocele; mala- 
ria; measles. 

Nueva Ecija: Cancer; malaria; rheumatism; tuberculosis. 


1x,B,3 Calderon: Medical Geography of the Philippines P11 


Nueva Vizcaya: Abscesses; bronchitis, acute; convulsion, infantile; furun- 
culosis; malaria; rheumatism; tuberculosis, pulmonary. 

Occidental Negros: Congestion, pulmonary; coryza; debility, congenital; 
dysentery; gastroenteritis in children; malaria; parasites, intestinal; 
pemphigus; psoriasis (of mountain people); pulmonary diseases in 
children; trachoma; tuberculosis; ulcers. 

Oriental Negros: Adenitis; anemia; blennorrhagia; bronchitis; broncho- 
pneumonia; chlorosis; convulsion, infantile; dactylitis; dysentery, bacil- 
lary; eclampsia; epilepsy; erysipelas; epithelioma; fever, septic; fever, 
typhoid; fibroma; fistula; gastroenteritis; grippe; hematocele; hemor- 
rhoids; hernia, inguinal; hydrocele; hysteria; jaundice; lipoma; lupus; 
malaria; measles; meningitis; necrosis; nephritis; neuralgia; onychia; 
parasites, intestinal; polyps; rheumatism; scurvy; tetanus; tonsillitis; 
tuberculosis, pulmonary; ulcers. 

Palawan: Dysentery; malaria; skin diseases; tuberculosis. 

Pampanga: Abscesses; adenitis; anthrax; beriberi, infantile; bone diseases, 
tubercular and syphilitic; bronchitis; bronchopneumonia; calculus; 
convulsions; dengue; dysentery; enteritis; grippe; malaria; measles; 
meningitis; parasites, intestinal; rheumatism; tuberculosis, pulmonary; 
typhoid; varicella; yaws. 

Rizal: Anthrax; beriberi; convulsion, infantile; dysentery; enteritis; furun- 
culosis; malaria; phlegmon; typhoid. 4 

Romblon: Diseases, cardiorenal; diseases, cardiovascular; dysentery; ma- 
laria; tuberculosis, pulmonary; typhoid. : 

Samar: Malaria; syphilis; tuberculosis. 

Sorsogon: Anthrax; bronchitis; bronchopneumonia; convulsion, infantile; 
dactylitis; dysentery; enteritis; fever, typhoid; fevers, eruptive; furun- 
culosis; gastritis; grippe; malaria; measles; phlegmon; rheumatism; 
septicemia, puerperal; tuberculosis; ulcers (foot and leg); yaws. 

Tarlac: Convulsion, infantile; malaria; tuberculosis. 

Tayabas: Beriberi; bronchitis; diseases, uterine; dysentery; eclampsia, in- 
fantile; gastroenteritis; goiter, simple; herpes; malaria; pneumonia; 
rheumatism; tetanus, umbilical; tuberculosis; whooping cough. 

Zambales: Convulsion, infantile; dysentery, bacillary; gastroenteritis; ma- 
laria; tuberculosis. 


TABLE II.—Monthly distribution of diseases in the Philippine Islands. 


Albay: 
January: Bronchitis; eclampsia, infantile; rheumatism; tuberculosis. 
February: Bronchitis; eclampsia, infantile; rheumatism; tuberculosis. 
March: Bronchitis; dysentery; eclampsia, infantile; rheumatism; 
tuberculosis. 

April: Eclampsia, infantile; tuberculosis. 

May: Dysentery, epidemic; eclampsia, infantile; malaria; tuberculosis. 
June: Dysentery, epidemic; eclampsia, infantile; grippe; malaria; 


tuberculosis. 

July: Dysentery, epidemic; eclampsia, infantile; grippe; malaria; 
tuberculosis. 

August: Dysentery, epidemic; eclampsia, infantile; grippe; malaria; 
tuberculosis. 


212 The Philippine Journal of Science 1914 


Albay—Continued. 
September: Bronchitis; dysentery, epidemic; eclampsia, infantile; ma- 
laria; tuberculosis. 
October: Bronchitis; dysentery; eclampsia, infantile; malaria; tuber- 


culosis. 
November: Bronchitis; dysentery; eclampsia, infantile; malaria; tuber- 
culosis. : : 


December: Dysentery; eclampsia, infantile; malaria; tuberculosis. 
Batangas: 
January: Beriberi, infantile; bronchitis; dengue; laryngitis. 
February: Beriberi, infantile; bronchitis; dengue; laryngitis. 
March: Bronchitis. 
April: Gastroenteritis; parasites, intestinal. 
May: Dysentery; enteritis; gastroenteritis; malaria; parasites, intes- 
tinal. 
June: Dysentery; enteritis; gastroenteritis; malaria; parasites, intes- 
tinal. 
July: Dysentery; enteritis; gastroenteritis; parasites, intestinal. 
August: Dysentery; enteritis; malaria. 
September: Malaria. 
October: Malaria. , 
November: Beriberi, infantile; dengue; laryngitis; malaria. 
December: Beriberi, infantile; bronchitis; dengue; laryngitis. 
Bohol: 
January: Bronchial diseases; coryza. 
July: Dysentery, amebic. 
August: Dysentery, ameebic; grippe; rheumatism. 
September: Grippe; rheumatism. 
October: Grippe; rheumatism. 
December: Bronchial diseases; coryza. 
Bulacan: 
January: Bronchitis; malaria; tuberculosis, pulmonary. 
February: Bronchitis; malaria; tuberculosis, pulmonary. 
May: Dysentery; gastroenteritis. 
June: Beriberi, infantile; dysentery; gastroenteritis. 
July: Beriberi, infantile; bronchitis; dysentery; gastroenteritis. 
August: Beriberi, infantile; bronchitis; dysentery; gastroenteritis. 
September: Beriberi; bronchitis; dysentery. 
October: Beriberi; bronchitis. 
November: Malaria; tuberculosis, pulmonary. 
December: Bronchitis; malaria; tuberculosis, pulmonary. 
Capiz. 
January: Dengue; dysentery; enteritis; tuberculosis; measles; vari- 
cella. 
February: Dengue; dysentery; enteritis; tuberculosis; measles; vari- 
cella. 
March: Dengue; dysentery; enteritis; tuberculosis; measles; varicella. 
April: Dengue; dysentery; enteritis; tuberculosis; measles; varicella. 
May: Dengue; dysentery; tuberculosis; measles; varicella. 
June: Dengue; dysentery; tuberculosis; measles; varicella. 
July: Bronchitis; malaria; rheumatism. 
August: Bronchitis; malaria; rheumatism. 


1x,B,3 Calderon: Medical Geography of the Philippines 913 


Capiz—Continued. 
September: Bronchitis; malaria; rheumatism. 
October: Bronchitis; malaria; rheumatism. 
November: Bronchitis; malaria; rheumatism. 
December: Bronchitis; malaria; rheumatism. 
Cavite: 
January: Bronchitis; malaria. 
February: Malaria. 
March: Dengue; dysentery; enteritis; tuberculosis; measles; varicella. 
April: Dengue; dysentery; enteritis; tuberculosis; measles; varicella. 
May: Dengue; dysentery; tuberculosis; measles;. varicella. 
June: Dysentery; malaria. 
July: Bronchitis; malaria; rheumatism. 
August: Bronchitis; dysentery; malaria. 
September: Bronchitis; malaria; rheumatism. ‘ 
October: Dysentery; malaria. 
November: Malaria. 
December: Bronchitis; malaria. 
Cebu: 
January: Dysentery; enteritis, acute; fever, intermittent; meningitis, 
simple. 
February: Enteritis, acute; fever, intermittent; meningitis, simple. 
March: Enteritis, acute; meningitis, simple. 
April: Dengue; dysentery; enteritis; tuberculosis; measles; varicella. 
May: Dengue; dysentery; tuberculosis; measles; varicella. 
June: Dysentery; malaria. 
July: Enteritis, acute; grippe; meningitis, simple. 
August: Bronchitis; dysentery; malaria. 
September: Bronchitis; malaria; rheumatism. 
October: Enteritis, acute; meningitis, simple. 
November: Dysentery; enteritis. 
December: Bronchitis; malaria. 
Tlocos Sur: 
March: Measles; varicella. 
April: Measles; varicella. 
May: Dysentery; gastroenteritis. 
June: Dysentery; gastroenteritis. ° 
July: Dysentery; gastroenteritis. 
November: Malaria; pulmonary diseases; rheumatism. 
December: Malaria; pulmonary diseases; rheumatism. 
lloilo: 
February: Measles. 
March: Measles. 
April: Measles. 
May: Dysentery; enteritis. 
June: Dysentery; enteritis. 
July: Dysentery; enteritis. 
August: Dysentery; enteritis; malaria. 
September: Dysentery; enteritis; malaria. 
October: Dysentery; enteritis; malaria. 
November: Dysentery; enteritis; malaria. 
December: Malaria. 


214 The Philippine Journal of Science 1914 


Isabela: 
January: Malaria. 
February: Malaria. 
March: Malaria. 
April: Malaria. 
May: Malaria. 
June: Malaria. 
July: Malaria. 
August: Malaria. 
September: Malaria. 
October: Malaria. 
November: Malaria. 
December: Malaria. 
Jolo: 
July:, Dysentery; malaria. 
August: Dysentery; malaria. 
September: Dysentery; malaria. 
October: Dysentery; malaria. 
November: Dysentery; malaria. 
December: Dysentery; malaria. 
Laguna: 
January: Convulsions, infantile; dysentery. 
February: Bronchopulmonary diseases; convulsions, infantile; dys- 
entery. 
March: Bronchopulmonary diseases; convulsions, infantile; dysentery. 
April: Bronchial diseases; dysentery; malaria. 
May: Bronchopulmonary disorders; dysentery; enteritis; malaria. 
June: Bronchopulmonary diseases; dysentery; enteritis; malaria. 
July: Bronchopulmonary diseases; dysentery; enteritis; malaria. 
August: Bronchopulmonary diseases; dysentery; enteritis; malaria. 
September: Bronchopulmonary diseases; convulsions, infantile; dysen- 
tery. 
October: Convulsions, infantile; dysentery. 
November: Convulsions, infantile; dysentery. 
December: Convulsions, infantile; dysentery. 


La Union: 
August: Beriberi; dysentery; fevers, catarrhal; gastrointestinal dis- 
orders. } 
September: Beriberi; dysentery; fevers, catarrhal; gastrointestinal 
disorders. 
October: Beriberi; dysentery; fevers, catarrhal; gastrointestinal dis- 
orders. : 
November: Beriberi; dysentery; fevers, catarrhal; gastrointestinal dis- 
orders. 
December: Beriberi; dysentery; fevers, catarrhal; gastrointestinal dis- 
orders. 
Leyte: 


January: Malaria; pulmonary diseases; scabies. 
February: Malaria; scabies. 

March: Dysentery; measles. 

April: Dysentery; measles. 


s dn 


1x,B,3 Calderon: Medical Geography of the Philippines 215 


Leyte—Continued. 
May: Diarrhea; dysentery; measles. 
June: Enteritis. 
July: Enteritis. 
August: Enteritis. 
November: Pulmonary diseases. 
December: Malaria; pulmonary diseases. 
Mindanao: F 
January: Bronchitis; dysentery; gastroenteritis; malaria; measles; 
rheumatism, articular; typhoid. 
February: Bronchitis; dysentery; gastroenteritis; malaria; measles; 
rheumatism, articular; typhoid. 
March: Bronchitis; dysentery; gastroenteritis; malaria; measles; rheu- 
matism, articular; typhoid. 
April: Malaria. 
May: Dysentery; gastrointestinal diseases; malaria; parasites, in- 
testinal. 
June: Dysentery, bacillary and amebic; malaria. 
July: Dysentery, bacillary and amecebic; malaria. 
August: Dysentery, bacillary and ameebic; malaria. 
September: Dysentery; malaria. 
October: Dysentery; malaria. 
November: Dysentery; malaria. 
December: Dysentery; malaria. 
Mindoro: 
January: Malaria. 
February: Malaria. 
March: Malaria. 
April: Malaria. 
May: Malaria. 
June: Malaria. 
July: Malaria. 
August: Malaria. 
September: Malaria. 
October: Malaria. 
November: Malaria. 
December: Malaria. 
Nueva Ecija: 
January: Malaria. 
February: Malaria. 
March: Malaria. 
April: Malaria. 
May: Malaria; typhoid. 
June: Malaria; typhoid. 
July: Bronchopulmonary diseases; rheumatism. 
August: Bronchopulmonary diseases; rheumatism. 
September: Pulmonary diseases; rheumatism. 
October: Pulmonary diseases; rheumatism. 
November: Malaria; rheumatism. 
December: Malaria; rheumatism. 
1263842 


216 The Philippine Journal of Science 1914 


Nueva Vizcaya: 
January: Bronchitis, acute; malaria. 
February: Malaria; rheumatism, articular. 
March: Malaria; rheumatism, articular. 
August: Malaria. 
November: Bronchitis, acute; rheumatism, articular. 
Occidental Negros: 
March: Gastroenteritis in children. 
April: Gastroenteritis in children. 
May: Malaria; gastroenteritis in children. 
June: Malaria. 
July: Malaria. 
August: Malaria. 
September: Malaria. 
October: Malaria. 
December: Coryza. 
Oriental Negros: 
March: Dysentery; gastroenteritis; measles; smallpox; typhoid. 
April: Dysentery; gastroenteritis; measles; smallpox; typhoid. 
May: Dysentery; gastroenteritis; measles; smallpox; typhoid. 
June: Dysentery, bacillary; gastroenteritis in children; malaria. 
July: Dysentery, bacillary; gastroenteritis in children; malaria. 
August: Dysentery, bacillary; gastroenteritis in children; malaria. 
September: Dysentery, bacillary; gastroenteritis in children; malaria. 
October: Malaria. 
November: Bronchitis; dysentery; grippe; malaria; whooping cough. 
December: Bronchitis; dysentery; malaria; whooping cough. 
Palawan: 
May: Malaria. 
June: Malaria. 
July: Malaria. 
August: Malaria. 
September: Malaria. 
Pampanga: 
January: Malaria. 
February: Dysentery; malaria. 
March: Dysentery; measles; varicella; typhoid. 
April: Convulsions; dysentery, bacillary; malaria; measles; typhoid; 
varicella. 
May: Convulsions; dysentery, bacillery; malaria; measles; varicella. 
June: Bronchitis; convulsions; dysentery, bacillary; gastroenteritis; 
malaria; measles; varicella. 
July: Bronchitis; gastroenteritis; measles; rheumatism; varicella. 
August: Beriberi; bronchitis; gastroenteritis; rheumatism. 
September: Beriberi; rheumatism. 
October: Beriberi; malaria; rheumatism. 
November: Malaria; rheumatism. 
December: Malaria; rheumatism. 


: 


1x,B,3 Calderon: Medical Geography of the Philippines P17 


Rizal: 


January: Anthrax; furunculosis; phlegmon; typhoid. 
February: Anthrax; furunculosis; phlegmon; typhoid. 
March: Typhoid. 

April: Typhoid. 

August: Dysentery; enteritis. 

September: Dysentery; enteritis. 

October: Dysentery; enteritis. 

November: Typhoid. 

December: Anthrax; furunculosis; phlegmon; typhoid. 


Romblon: 


January: Dysentery; measles. 
February: Dysentery; measles. 
March: Dysentery; measles. 
October: Malaria. 

November: Malaria. 
December: Malaria. 


Samar: 


March: Tuberculosis. 

April: Tuberculosis. 

May: Malaria; tuberculosis. 

June: Malaria; syphilis; tuberculosis. 
July: Malaria; syphilis; tuberculosis. 
August: Malaria; syphilis; tuberculosis. 


Sorsogon: 


February: Diarrhoea; dysentery; fever, typhoid; fevers, eruptive. 

March: Diarrhoea; dysentery; fever, typhoid; fevers, eruptive. 

April: Diarrhcea; dysentery; fever, typhoid; fevers, eruptive. 

June: Bronchitis; malaria; pleurisy; pneumonia; rheumatism; tonsil- 
litis. 

July: Bronchitis; malaria; pleurisy; pneumonia; rheumatism; tonsil- 
litis. 

August: Bronchitis; malaria; pleurisy; pneumonia; rheumatism; tonsil- 
litis. 

September: Bronchitis; malaria; pleurisy; pneumonia; rheumatism; 
tonsillitis. 

October: Bronchitis; malaria; pleurisy; pneumonia; rheumatism; ton- 
sillitis. 


Tarlac: 


June: Dysentery. 
July: Dysentery. 
August: Dysentery. 
September: Dysentery. 


Tayabas. 


August: Bronchitis; malaria; tuberculosis; typhoid. 
September: Bronchitis; malaria; tuberculosis; typhoid. 
October: Bronchitis; malaria; tuberculosis; typhoid. 
November: Bronchitis; malaria; tuberculosis; typhoid. 
December: Bronchitis; malaria; tuberculosis; typhoid. 


218 The Philippine Journal of Science 1914 


Zambales: 
January: Tuberculosis. 
February: Tuberculosis. 
March: Tuberculosis. 
April: Tuberculosis. 
May: Tuberculosis. 
June: Tuberculosis. 
July: Dysentery; tuberculosis. 
August: Dysentery; tuberculosis. 
September: Tuberculosis; dysentery. 
October: Tuberculosis. 
November: Tuberculosis. 
December: Tuberculosis. 


THE ETIOLOGY OF TRICHOMYCOSIS PALMELLINA IN THE 
PHILIPPINE ISLANDS 


By Otto ScHOBL 
(From the Biological Laboratory, Bureau of Science, Manila, P. I.) 


One plate 


This pathological condition of hair was first described by F. J. 
Pick in 1875,(1) who considered it to be of microbial origin. 
It is most frequently restricted to the axillary regions. The 
disease presents itself as a more or less marked thickening of 
the individual hair. In the early stage, small nodular thicken- 
ings of whitish color are to be found on the hair. In the 
later stage, the thickenings of the hair become confluent and 
a sheath, evidently of foreign matter, frequently surrounds the 
hair along its entire length. At this stage, it is more likely 
to be pigmented than in the early stage. Pasty matter can 
be easily scraped off the diseased hair. The hair tears easily, 
and its epilation is scarcely felt by the patient. Partial and 
temporary loss of hair in the affected region may occur. Besides 
the typical location in the axillary grooves, the condition has 
been found in the pubic region. One case of trichomycosis 
palmellina capilitii was observed by Winternitz. (2) 

The etiology of the disease being early recognized as microbial, 
investigations were commenced to identify the particular micro- 
organisms, as well as experimentally to support the theory that 
the bacteria found to be present on the diseased hair were the 
actual cause of the condition and not merely casual saprophytes 
as several authors had claimed. It was not until thorough 
bacteriological examinations were made that some light was 
thrown on the question. 

Behrend (38) found a coccus which liquified gelatin, and formed 
white and yellow colonies when cultivated on agar plates. 
Eisner (4) describes a Gram-positive coccus which he isolated 
from a case of trichomycosis. As this coccus showed cultural 
characteristics similar to those described by Behrend, Hisner 
considers both microbes as identical. Similar findings were 
made by Sonnenberg (5) and Colombini (5) the latter author 
being successful in transmitting the disease. Winternitz(2) found 
in a case of trichomycosis palmellina capilitii a nonmotile bacillus 
which showed chromatic granules and formed pronounced involu- 
tion forms in old cultures. The author believes his bacillus to be 
different from the microbe isolated from trichorhexis nodosa 
by Hodara and Spiegler. (6) 

219 


220 The Philippine Journal of Science 1914 


The disease being evidently common among Caucasians living 
in the Philippine Islands, it seemed of interest to compare the 
bacteriological findings made in Manila with those made else- 
where, particularly in temperate climates, especially as the 
bacteria found to be connected with the condition in question 
were not sufficiently classified to warrant their identification. 
Furthermore, it is evident from the brief outline of previous 
examinations that the findings made by the various authors do 
not refer to identical microbes. Therefore, it remains doubtful 
whether or not trichomycosis palmellina as a disease is of one 
etiology. 

It is the consensus of opinion that the disease occurs more 
frequently in blonds than in brunets. The pigmentation of 
the skin seems to be of significance, as the albinos of the 
dark races are also susceptible. (Compare Wisner’s(4) case 
of an albino Negro showing trichomycosis palmellina.) This rule 
evidently holds true also in the tropics as the disease seems 
to be absent among the natives. Still other factors must be 
taken into consideration, such as the lack of hair in the places 
of predilection among the Orientals, as well as the increased 
perspiration of the whites in the tropics as compared with 
the colored races of the Orient. There is no doubt that perspi- 
ration causes the seasonal occurrence and recurrence of the 
disease. During the cooler months or during the sojourn in a 
region of high altitude it was noticed that the disease was 
reduced to a minimum, while during the hot season the disease 
reached its climax, sometimes in spite of scrupulous cleanliness. 
In one case under observation, folliculitis with corresponding 
lymphadenitis was observed. Staphylococci were evidently a 
secondary invader in this instance. 


MICROSCOPICAL EXAMINATION 


The foreign matter which causes the thickening of the diseased 
hair was scraped off by means of a sterile scalpel. Smears 
were prepared therefrom and stained by the usual methods; 
unstained preparations were also examined. The material was 
found to be composed of bacteria clumped in zodgloeal masses. 
The great majority of the organisms were Gram-positive, non- 
motile, rather short bacilli of the Corynebacterium type; that is, 
rod-shaped bacteria with rounded ends, one end of the rod being 
thicker than the other. Elongated forms were also present. 
Chromatic granules were evident in smears stained with Loffler’s 
methylene blue. Large cocci of the staphylococcic type were 
also found. They were more numerous in the latter stage of 


IX, B, 3 Schobl: Etiology of Trichomycosis Palmellina 291 


the disease, while in the beginning very few were found or none 
at all. The cocci showed no tendency to form zoéglea. The 
examination of an unstained preparation revealed no motile 
organisms, although Brownian movement was very pronounced. 


MICROSCOPICAL EXAMINATION OF THE DISEASED HAIR IN TOTO 
STAINED AND UNSTAINED 


Unstained preparation.—The diseased hair was placed on a 
slide, suspended in a drop of salt solution, and examined micro- 
scopically under a cover glass. The foreign matter was found to 
be scattered along the shaft of the hair in isolated spots or in the 
latter stage of the disease surrounding the whole length of the 
shaft, but leaving always the root and, as arule, the tip of the hair 
free. It consists of transparent finely granular matter, the 
outline of the hair being visible within the granular masses. 
Under high power, distinct nonmotile rod-shaped bacteria were 
distinguishable on the periphery of the granular aggregations. 

Stained preparation.—The method of Hodara and Spiegler was 
employed. The hair was first treated with ether to free it 
from fat, then bleached with peroxide of hydrogen, and stained 
by Gram’s method followed by counterstaining with Van Gie- 
son’s stain. 

On account of the double stain, the bacteria being stained 
blue in contrast to the yellow-stained hair, the conditions of 
the hair as already described were much more conspicuous. 
Small groups of not over 20 bacteria could be seen scattered 
among the large microbial aggregations. The outside layer 
of the hair was split off in several places, and streaks of bacteria 
could be followed into the deeper layers of the hair. This 
anatomical change of the hair, which could be noticed on 
specimens untreated with chemicals, would seem to corroborate 
the theory that mechanical injury precedes the infection. 

Repeated examinations of specimens, taken from time to time 
from the same patient, revealed all the stages of the pathological 
process under question. Small groups of bacteria could be 
found on the still intact cuticula of the hair, while the next 
focus showed defects of cuticula and bacteria penetrating in 
the substantia pili. In the latter stage, masses of bacteria 
were found in the intercellular spaces of the cortical substance. 
This led to splitting of the hair and to its final disintegration. 
It is evident that the effect of the bacterial invasion upon the 
hair is purely mechanical. Primarily, the bacterial growth takes 
place on the surface of the hair. After the cuticula has been 
destroyed the bacteria grow in the preformed cavities of the 


222. The Philippine Journal of Science 1914 


substantia corticalis. Against the invasion of the bacteria the 
hair is defenseless, not being equipped with direct blood supply 
and hence lacking all means of defence against infection. The 
anatomical conditions of the radix pili are different from those 
of the shaft.(6) The cellular elements of the root of the hair 
being less differentiated than those of the shaft, the intercellular 
spaces are solid and the blood supply is nearer at hand. Hence 
the radix pili remains normal. Needless to say, mechanical 
injury of the hair followed by loss of the cuticula of the hair 
might facilitate the bacterial invasion, although the scalelike 
arrangement of the cornified cuticular cells would possibly allow 
the bacteria to penetrate without injury into the hair substance. 
At any rate, the injury might be of minimal extent to bring 
about the loss of the cuticula. 


EXAMINATION BY CULTURES 


The diseased hair was epilated by means of sterile forceps, 
and streak culture was made on agar plates. The cultures were 
incubated at 37°C. The cultural findings as compiled from 
repeated examinations are as follows. 

Index of cultures found in cases of trichomycosis palmellina. 
I. Bacteria found constantly in any stage of every case. 
1. Small round granular colonies of Corynebacterium. 
2. Small umbilicated coarsely granular flat colonies, with irregular 
margin, of Corynebacterium. 
II. Bacteria found frequently: 
3. Large round homogeneous white colonies of a large micrococcus. 
4, Large round lemon-yellow-colored colonies of a large micrococcus. 
III. Bacteria found occasionally: 
5. Large white umbilicated colonies of a large micrococcus. 
6. Orange-yellow soft colonies of a small micrococcus. 
7. Round concentric olive-green colonies of Sarcina. 
8. Round faint yellow-colored colonies of a micrococcus. 
9. Colonies of a staphylococcus resembling Staphylococcus aureus. 


TABLE I.—Distribution of bacteria in the cases studied. 


Bacteria culture 
| Case. No. 


1 0 BS cae ae 2 1, 2, 8, and 4. 
255 Bo SD sete 1, 2, 4, 5, and 9. 
Bok. Wee 1, 4, and 5. 
ASMoM 202-825 1, 2, 3, 4, and 6. 
Nil D ee ee ae 1, 3, and 7. 

Ch a ae | 1, 2, 3, and 6. 
Mee = 2 Sema 1, 2, 3, 5, 6, and 7. 
Bdeark’ 22 520-=- 1, 2, and 3. 


T/L one ae 1, 3, 4, 7, and 8. 
10) Gree. ues - = 1, 3, and 4. 


1X, B, 8 Schobl: Etiology of Trichomycosis Palmellina 293 


It is evident from bacteriological examinations of ten cases 
of trichomycosis as summarized in the table that bacteria 
from the pseudodiphtheria group were found in every case to 
predominate over the rest of the bacterial flora. In the early 
stage, these organisms were present in practically pure culture. 
Two types of colonies were encountered, the morphology of the 
organisms being practically the same. The phenomenon of 
mutation was thought of, and great care was taken to secure 
pure subcultures of the two varieties of colonies. The types 
of colonies being indistinguishable by the naked eye in young 
cultures, Barber’s method for isolation of a single organism 
was applied in order to secure guaranteed pure cultures. This 
method as applied to transplanting minute colonies from plates 
being done under the low power of the microscope, any possible 
contamination due to the inclusion of one colony by another 
could be safely avoided. I have failed to ascertain any change 
in the shape of colonies in pure cultures obtained in this way. 

The cultures of the pseudodiphtheria group which were isolated 
from the cases of trichomycosis acidified glucose, maltose, and 
saccharose. Dextrin, inulin, galactose, mannite, raffinose, ery- 
thrite, inosite, and dulcite remained unchanged. The acidifica- 
tion was very slight, and occurred after several days’ incubation. 
Gelatin was not liquified. In bouillon, the growth occurred at the 
bottom of the tube in the shape of a flaky sediment. Deep 
agar stab culture on top of which melted agar was poured 
developed scanty growth. On agar slant cultures, the growth 
of isolated colonies was of the shape already described. Agar 
stab culture showed arborescent growth. No hemolysis occurred 
on human blood agar. White mice survived subcutaneous in- 
oculation of 1 cubic centimeter of a bouillon culture which was 
several days old. 

The bacterium second in frequency found in trichomycosis was 
a large micrococcus forming white colonies and an organism 
morphologically identical with the former but forming yellow 
colonies (Nos. 3 and 4). These micrococci correspond to the 
description of the organism found by others as far as mor- 
phology and growth on the usual culture media are concerned. 

The large white coccus acidified glucose, maltose, and raffinose. 
Mannite, dextrin, dulcite, amygdalin, galactose, and lactose 
remained unchanged. The large yellow coccus acidified glucose 
and saccharose. The rest of the sugars mentioned above 
remained unchanged. 

Besides the bacteria already mentioned, other chromogenic 
organisms were found. Their number was very limited, as a 


224 The Philippine Journal of Science 1914 


rule two or three colonies on a plate. They are marked in the 
index of cultures by numbers 5, 6, 7, 8, and 9. It is more than 
probable that these bacteria are mere casual contaminations from 
the skin and play no part in the condition under question. 

As the disease is so widespread among the white people 
in the Philippine Islands that 10 out of 11 whites picked at 
random showed well-developed trichomycosis, it was thought use- 
less to attempt transmission experiments on man, on account 
of the objection that the person chosen for such experiment 
already had the disease at the time of inoculation. Indeed, at 
times the condition might be of such a minute extent that the 
hair appears to be normal, but microscopical examination, par- 
ticularly that of the stained hair in toto, reveals its presence. 
Recourse was, therefore, had to experiments in vitro. Hairs 
that were found by microscopic examination to be perfectly 
healthy were placed in tubes of bouillon and sterilized; the tubes 
remained sterile after an incubation period of two days. Various 
cultures isolated from different cases of trichomycosis were 
planted in the bouillon containing the hairs and incubated at 
37°C. The experiments were not successful. 

The constant presence of Corynebacterium and its prevalence 
in the flora of trichomycosis palmellina in every stage of the 
disease, as found by repeated microscopical examination and 
by cultures, as well as the direct connection of these bacteria 
with the lesions of the diseased hair, as seen on microscopical 
examination of the hair in toto, leaves but little doubt that, what- 
ever the primary “causa lesionis” may be, the above-mentioned 
class of bacteria, which are so widespread and so commonly 
found on the skin and surface mucous membranes, are responsible 
for the pathological condition known as trichomycosis palmellina. 


REFERENCES 


(1) Pick. Vierteljahrschrifte fiir Dermatologie und Syphilis (1876), 3, 
625. 

(2) WINTERNITZ. Arch. f. Derm. u. Syph. (1908), 66, 81. 

(3) BEHREND. Berl. klin. Wochenschr. (1890), 27, 464. 

(4) E1rsner. Arch. f. Derm. u. Syph. (1897), 41, 59. 

(5) SONNENBERG and COLOMBINI. Referred to by Winternitz (2). 

(6) Sprecuer. Arch. f. Derm. u. Syph. (1897), 41, 67. 

(7) RauBer-KopscH. Lehrbuch der Anatomie. 8 Auflage (1914). 

(8) BarBer. Phil. Journ. Sci., Sec. B (1914), 9. 


a ae 


ILLUSTRATIONS 


PLATE I 


Zodgleal masses of bacteria forming a continuous sheath surround- 
; ing the hair. Case O.S. Zeiss Ocular 4 Obj. AA. 

2. The end of the hair where it was clipped off in vivo. Bacteria 
growing within the split hair. Case O. S. Zeiss Ocular 4 Obj. 


DD. 
ace section through the diseased hair. Bacteria penetrating 


i 


within the hair. Case O.S. “Homm. immer.” 2 Ocular 2. 
225 


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ScHOBL: TRYCHOMYCOSIS. ] [PHIL. JourRN. Sci., IX, B, No. 


Fig. 1. Zoogleal Fig. 2. Bacteria growing within 
masses of bacteria the split end of a hair. 
forming a continuous 
sheath surrounding 
a hair. 


o\ 


Fig. 3. Cross section through a diseased hair, showing bacteria penetrating within the hair. 


PLATE |. THE ETIOLOGY OF TRICHOMYCOSIS PALMELLINA. 


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A CONTRIBUTION TO THE BACTERIOLOGY OF LEPROSY 
PRELIMINARY NOTE 


By JoHN A. JOHNSTON 
(From the Biological Laboratory, Bureau of Science, Manila, P. I.) 


One plate 


Since Hansen (1) in 1872 announced his discovery of ‘the 
specific cause of this disease, many men in different parts of 
the world have endeavored to cultivate Bacillus lepre with varied 
success. Twenty-eight cultivations have been reported and 
cited,(2) of which 20 were organisms of the diphtheroid type, 
5 were rods, and 3 were Streptothrix. None of these organ- 
isms has received a general recognition as the real Bacillus 
lepre. Only two workers, Kedrowski and Bayon, report any 
results of animal inoculation with cultures that bear any patho- 
logical similarity to the lesions of human leprosy. Probably 
the earliest reported isolation was that of Bordoni-Uffreduzzi 
in 1888, who succeeded in growing an acid-fast diphtheroid 
organism. Since this date there has gradually crept into the 
literature, and now is apparently generally accepted, the term, 
acid resisting, for an organism which shows any tendency to 
hold stain against acids. Bacillus leprxe, as we find it in the 
tissues and juices of the body, is distinctly acid fast, although 
not quite so acid fast as the tubercle bacillus; but to term a 
bacillus, which, after staining by carbol fuchsin, readily de- 
colorizes when treated for a few seconds with 20 per cent nitric 
or 25 per cent sulphuric acid, Bacillus lepre is decidedly un- 
warranted. Excluding these so-called acid-resisting organisms, 
the reported cultivations of Bacillus lepre are reduced to 6. Of 
these organisms, 4 are of the diphtheroid type and 2 are rods. 

How may these discrepancies be explained? We know that 
in cultures of the tubercle bacillus a filamentous form of growth 
has frequently been observed. This has also been noted in 
sputum of tuberculous persons, where hyphzlike filaments are 
branched and often have swollen ends. In accordance with 
these findings, some bacteriologists class the tubercle bacillus 
with the trichomycetes or with the true molds of which the 

227 


228 The Philippine Journal of Science 1914 


streptothrix is a type, while others place the tubercle bacillus 
and closely allied organisms in a special group holding an inter- 
mediate position between the streptotricheze and the ordinary 
bacilli. I believe, with Jordan,(3) that the tubercle bacillus and 
its near allies will eventually prove to be parasitic forms of the 
higher molds. Foulerton,(4) in 1910, in his Milroy lectures 
before the Royal College of Physicians, London, mentions 

the apparent affinity between certain undoubted species of Streptothrix and 
the parasite, or parasites, of tuberculosis. A comparison of the biological 
characteristics of the recognized streptothrix organisms on the one hand 
and of different “strains” of the parasite of tuberculosis on the other 
leaves no doubt as to the correctness of the opinion held by certain earlier 
pathologists who maintained, within a few years of Koch’s announcement 
of his discovery of the cause of tuberculosis, that the reputed bacillus was 
not, in fact, a fission fungus at all, but rather belonged to a higher group 
of mold fungi. 

There is, undoubtedly, a very close relation between leprosy 
and tuberculosis, and if we accept Foulerton’s conclusions as 
correct in regard to tuberculosis why may not similar ones be 
true of the organism known as Hansen’s bacillus? 

From the spleens of two lepers who died at San Lazaro Hos- 
pital I have succeeded in cultivating an absolutely nonacid-fast 
streptothrix. These two strains are apparently identical, and 
now grow readily on the ordinary glycerin agar, but the original 
isolations were slow in growth. They were made on placental 
agar and fish-juice agar, and required from three to four weeks 
for growth to become apparent. The growth of this organism 
may be described as spreading, with a tendency toward the 
formation of small islets with an elevated center. These even- 
tually coalesce, and the surface growth is dull and more or less 
rugose. It is pearly white in young cultures, becoming brown 
in cultures more than four months old. There is a tendency 
to spore formation in old cultures, as evidenced by the forma- 
tion of white patches which appear at the uppermost part of 
the stroke and gradually extend downward to the butt of the 
tube. The growth is very adherent to the medium, and cannot 
be removed without bringing medium along with it. 

In bouillon with or without glycerin there is a very scanty 
growth, and this has a tendency to creep up the sides of the 
tube and also to collect at the bottom as a powdery sediment. 
Stained preparations at this stage show long and short threads 
with a well-marked tendency to branch as shown in Plate I, a. 
In older cultures, three to four months old, there is tendency for 
the filaments to break up into coccoid and rodlike forms as seen 


| 


1X, B, 3 Johnston: The Bacteriology of Leprosy 2239 


in Plate I, b. These are, at first, nonacid fast; after six to 
seven months a few will be noted as retaining the fuchsin 
slightly, others will not stain at all either with the original 
stain or the counterstain, and after a year there will be found 
scattered clumps of distinctly acid-fast bacilli occurring as 
isolated individuals and rods still inclosed in the parent hypha 
(Plate I, c). 

In the latter part of February, 1913, several 48-hour cul- 
tures of this streptothrix were rubbed up in a mortar and a 
number of guinea pigs and rabbits were inoculated with this 
suspension. The rabbits were all inoculated with 1 cubic centi- 
meter intravenously; the guinea pigs, with 0.5 cubic centimeter 
subcutaneously in each groin. Of this series, 1 animal died 
while 3 animals are still alive; the others, numbering 16, 
were killed at varying periods from a week up to six months. 
No lesions were discovered post mortem in the series of animals 
killed. The guinea pig dying in September, however, showed a 
slightly enlarged liver with a few scattered nodules on the under- 
surface. There was no glandular enlargement. Smears from 
the cut surface of several of the nodules showed no organisms, 
but many small rounded masses from 3 to 6 microns in greatest 
diameter were visible; these were distinctly acid fast. I have 
noted similar masses in the juice expressed from a leproma, in 
scrapings from a nasal ulcer in a leper, and also in old cultures 
of the streptothrix. Some of these bodies had frayed-out edges, 
and resembled blood plates as we sometimes see them. 

Nine days after, cultures made from these nodules showed in 
two tubes of placental agar a slight whitish growth of about 
the consistency of cream cheese. It was spreading, did not 
grow in the water of condensation, and at the end of four weeks 
covered the surface of the slant. Stained preparations from 
these cultures showed long and short rods which were distinctly 
acid fast. Growth in bouillon was slow, and mostly at the 
bottom; there was occasional pelicle formation. Marked club- 
bing occurred, as seen in Plate I, d, e, and f. In bouillon the 
bacillary forms show a tendency to lose the acid-fast property; 
this is regained, however, on transfer to either placental agar 
or Dorset egg plus 1 per cent glycerin. All of the clubbed 
forms are distinctly acid fast. 

In April a second series of animals was inoculated as before. 
One of the guinea pigs developed a pussy discharge from its 
left eye eight days after inoculation. Stained smears showed 
enormous quantities of acid-fast bacilli. Cultures from this 


230 The Philippine Journal of Science 1914 


guinea pig were all negative at the end of two months. This 
animal died some nine months later, and the liver, lungs, spleen, 
and some of the lymphatic glands showed marked nodular in- 
volvement. Smears from the different organs showed a few 
acid-fast bacilli. Sections were examined by Dr. B. C. Crowell 
of the Bureau of Science, who reported that the lesions would, 
ordinarily, be described as tubercular. Cultures made from the 
spleen on placental agar showed after two weeks a growth prac- 
tically identical with the first isolation. Efforts to identify the 
organisms isolated by means of agglutination and deviation of 
complement tests have been so far unsatisfactory. 

So far as I know, but two other investigators have injected 
a streptothrix and recovered an acid-fast rod; these were Bayon 
of London and Kedrowski of Moscow. Bayon also recovered a 
streptothrix after inoculating a rat with this acid-fast rod. At 
the present time I have a series of animals under observation to 
test whether or not the acid-fast rod becomes a streptothrix in 
the animal body. I have noted, however, a decided tendency 
for the cultures of the acid-fast rod form to throw back, as it 
were, and show decided streptothrix forms. In conclusion I 
can only say that at this stage of my work I am quite convinced 
that Bacillus lepre is but the acid-fast stage of a markedly pleo- 
morphic streptothrix. 


BIBLIOGRAPHY 


1. HANSEN, A. Norsk. Mag. f. Laegevidensk (1872), 2, 1. 

2. WOLBACH, S. B., and HoNeEIJ, JAMES A. Journ. Med. Research (1914), 
29, 367. 

3. JORDAN, E. O. A textbook of General Bacteriology. W. B. Saunders 
Company, Philadelphia and London. 3d ed. (1918), 347. 

4. FouLerTon, A. G.R. Lancet (1910), 1, 551, 626, 769. 


ILLUSTRATION 


(a) Hyphz, showing branching—commencement of breaking- 
down stage; (6b) coccoid and bacillary forms; (c) clump 
of acid-fast bacilli; (d, e, f) clubbed forms. 


231 


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Medi Bong 
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Leo OD salt 
eet aren pili 
Fhigynd badsA reriess 


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beneek, haodelaton UY by AU. : 


At SOR ay 


seve ttt | i an a i ee 
NW ie ie et 1% ste 


JOHNSTON, J. A. BACTERIOLOGY oF LEPROSY.] [Puim. Journ. Scr., 1X, B, No. 3. 


PLATE |. (a) HYPHA, SHOWING BRANCHING—COMMENCEMENT OF BREAKING-DOWN 
STAGE; (b) COCCOID AND BACILLARY FORMS; (c) CLUMP OF ACID-FAST BACILLI; 
(d, e, f) CLUBBED FORMS. 


fake ae 


Se 
‘. 


INTESTINAL HELMINTHIASIS IN THE PHILIPPINE ISLANDS AS 
INDICATED BY EXAMINATIONS OF PRISONERS UPON 
ADMISSION TO BILIBID PRISON, MANILA, P. I. 


By Davin G. WILLETS 
(From the Biological Laboratory, Bureau of Science, Manila, P. I.) 


The results obtained in examinations of inhabitants of the 
Philippine Islands for evidences of intestinal parasitism have 
shown that the percentage of persons infected with any one 
species of organism has varied considerably. For instance, 
infections with Trichuris and with hookworms have ranged 
from 6 to 88 and 8 to 54 per cent, respectively. The investiga- 
tions yielding these results have been made in distinctly circum- 
scribed localities and chiefly among laborers. An accurate idea 
of the average prevalence of intestinal helminthiasis in the 
Archipelago could be obtained only by examining persons from 
all parts of the Islands, of all ages, of both sexes, of all occupa- 
tions, of all social classes, and of all races. Opportunity for the 
examination of individuals from all parts of the Islands is 
offered more or less completely at Bilibid Prison. The inmates 
are, however, mostly of the lower classes of society and children 
are rarely admitted. The examinations here reported were made 
upon 7,843 adult males upon admission to the prison. All cases 
which had been in the institution previously were excluded 
from the series for the reason that some of their infections 
may have been contracted in the prison. The Filipinos of the 
series number 6,416; the Chinese, 1,427. About 300 females 
were examined, but this number is too small to permit their 
inclusion in the report. It was intended originally to extend 
this investigation over several years, in order that a large series 
of cases might be secured. The work has been interrupted 
permanently, however, and therefore a report is made of the 
results at hand with the realization that they must be interpreted 
_with caution. Three phases of helminthiasis among adult male 
Filipinos and Chinese in the Philippine Islands are considered. 
First, the average prevalence of helminthiasis in Filipinos; 
secondly, its geographic distribution among Filipinos; and, 
thirdly, its relative frequency among Chinese and Filipinos. 

233 


234 The Philippine Journal of Science 1914 


The conditions limiting the work were as follows: 

1. One cover-slip preparation was examined of the one spec- 
imen obtained from each case. 

2. The majority of the examinations were made by Filipino 
“trusties” under the supervision of various members of the 
biological laboratory, Bureau of Science (Drs. P. E. Garrison, 
Y. K. Ohno, F. B. Brown, Liborio Gomez, and D. G. Willets). 
The findings are, therefore, not as exact as they would have 
been if the examinations had been made by more experienced 
individuals. 

3. Examinations were made within seventy-two hours of 
admission. Since travel from sections of the Islands which are 
most remote from Manila practically never requires more than 
three weeks and since about one month after the date of infec- 
tion with the several species of parasites concerned is necessary 
for evidences of them to appear in the feces, it may be considered 
that the positive results obtained indicate infections which the 
individuals examined possessed when they started for the 
prison. Protozoan findings were excluded because they may 
have been contracted en route. 

4. None of the individuals examined had been an inmate 
of the prison previously. All of them were adult males. 

The results obtained are given in Tables I, II, III, and IV. 
They may be compared with those of similar investigations in 
the Philippines by consulting tables prepared by me.: In making 
comparison, it should be noted that the majority of the series 
of cases studied are composed of males and females, as well 
as adults and children of given communities. 

Garrison ? examined Bilibid prisoners, and a summary of his 
helminthic findings is given in Table V. The percentage of 
persons harboring Trichuris, Ascaris, and hookworms differs 
greatly in his series of cases and mine. 

The cause of these variations cannot be due to any great 
extent to the personal equation entering into the examinations, 
because the ova of Trichuris, Ascaris, and hookworms are easily 
recognized by one seeing them daily,? nor can it be due to any 
change outside of the prison, for a greater percentage of the ad- 
mission cases of 1910 were infected than those of 1908. It is 
probable, therefore, that the variations are due to conditions 
within the prison at the time Garrison made his examinations 


1This Journal, Sec. B (1911), 6, 77. 
*Tbid. (1908), 3, 1911. 
* For instance, the “trusties” already mentioned. 


1X, B, 3 Willets: Intestinal Helminthiasis 235 


or prior thereto and that these conditions were favorable to 
the propagation of hookworms and Trichuris and unfavorable 
to that of Ascaris. The differences in the Trichuris and Ascaris 
percentages, however, may be explained in part by the fact 
that Ascaris infections are treated at the prison hospital as a 
routine procedure, whereas Trichuris infections are not treated 
unless they are heavy. Such a state of affairs would naturally 
lead to carelessness in observing light Trichuris infections in 
making examinations to detect cases for treatment, such as 
those here recorded. Garrison examined some admission cases 
but chiefly those which were already in the institution, and his 
findings represent an endemic condition.* Improved sanitary 
conditions in the institution have been efficacious in markedly 
lowering the incidence of intestinal helminthiasis as shown by 
the results obtained in the examinations of 930 prisoners in 
1910 (Table VI). As indicated in the table, these individuals 
had been prisoners for periods varying from a few months to 
four years. The only species of helminthic parasites which gave 
an increased percentage upon length of residence was the seat- 
worm (Oxyuris). 

A map showing the distribution of infections with the several 
species of parasites found in the examinations of the Filipinos 
of the series was prepared. It showed that the distribution 
of the various parasites is extremely irregular. Adjacent north- 
ern and adjacent southern provinces gave in some instances 
very different results. Former investigations predicted this 
result. Indeed, it is on record that Garrison, Leynes, and 
Llamas ° and Rissler and Gomez °® found quite different percent- 
ages of infection with Trichuris in Rizal Province. 

The low percentage of Chinese infected in general and with 
each common species of parasite, as shown in Tables II and 
III, is remarkable. The explanation of the differences in infec- 


‘It is well known that the sanitary conditions at the prison were deplor- 
able prior to their being placed in the hands of the Bureau of Health, 
shortly before Garrison’s investigation was begun. It is further known 
that these conditions were changed, so that within a period of a few months 
sound sanitary regulations were in force. Since that time, routine examina- 
tion of stools for evidences of intestinal parasitism has been regularly made 
by the Bureau of Science, and infected cases have been treated. Prisoners, 
new ones or those who have been detailed for outside work, are held in 
quarantine for five days upon admission. If an individual is found to 
harbor intestinal parasites other than Trichuris or monads, he is treated in 
the prison hospital until apparently cured. 

*Tbid. (1909), 4, 257. 

*Ibid. (1910), 5, 267. 


236 The Philippine Journal of Science 1914 


tion percentage among the Chinese and Filipinos must be that 
the former are exposed less than the latter to the sources of 
infection with intestinal helminthic parasites. It is to be noted 
in this connection (1) that Chinese use chopsticks when eating, 
whereas lower class Filipinos use their fingers; (2) that Chinese 
drink a great deal of tea (boiled water), whereas Filipinos 
do not; (8) that Chinese eat less uncooked food than Filipinos; 
(4) that many of the Chinese were shopkeepers or clerks, whereas 
most of the Filipinos were laborers, and consequently the former 
were less liable to soil infection than the latter. 


TABLE I.—Summary. 


| 
Examinations and infections. | Number. | Per cent. 


| *Personia"exam ined oe ee eee ee eee een ere een See ee oe eres H (ict 4 jl hontai Bac 
Perbona infected sso225 36. 2 ee Se ES ee ee ee eee 5, 421 69.1 
Persons infected with— a Tra 
RP RCBA US: oe ce ten SO a es Me ie Pee a ee St 3,690) 47.1 
A SCUTAS Soe Se ee ee ee ee ee ee ee eee eee eee eee 3, 211 40.9 
Hookwormi3 42. = ees! oot. ie ee ee ee eee 1, 737 22.2 
Stronquloides.. 2a = es ok OC ie, CS eh eee 65 0.8 
Ona Urt eo a oi a a ecb a fe ae ee reer 5T 0.7 
Tse ryil Soe EE Sa ee ee ee ee ee eee 57 0.7 
Clonorchisige Mes acne eS ee ee Sees 8 0.1 
ETOP ONGLE TAN FURIE Be ae te 8 I ek oe 4 0.05 
SCHISLOSONI JODONMIOCWIN = ~~ nn foe ee eee oe sth 8 ie ta Dah 3 0. 05 
Total no Fe ease tee ee ee ee ae ed ew oe Oe 8,832 | 112.60 
Soper TL is SAP EE z mt ete Se 


TABLE II.—Race distribution. 


Race. 
Total. 


Number. | Per cent.; Number. | Per cent.. Number. | Per cent. 


7: 
| 

Examinations and infections. | Filipinos. Chinese. 
| 


i 


ica ined .si04 5 eee <Bae See | MG ANG) eee Soe iP ate ADT, |Nee Skee 1,943) |5c_ 2 teen 


MORTHVE) eo-2- oo oe oe eS ee | 4,940 77.0 481 33.7 5, 421 69.1 
TROT TLS ee ene ec ee 3, 447 | 53.7 243 | 17.0 | 3,690 47.1 
ASCOrtR 2m. Cece. Seba eee ot | 2, 946 45.9 | 265 | 18.6 3, 211 40.9 
Hookworm -2-en ss soe ee sore | 1,648 24.0 94 6.6) 1,787 22.5 
SETORGUIOULES ee nee ee 60 0.9 | 5 0.4 65 0.8 
ORG UTE TT ere ene ae 55 0.9 2 0.1 57 0.7 
Vania... Sern ees SS 53 0.8 4 0.3 57 0.7 
Miscellaneous -..-2.-4-2-2-4--.-- 10 0.2 | 5 0.4 15 0.2 

pt | | nn | 
Total infections’ =-----+---2=-=- 8, 214 128.0 618 43.3 8, 832 112.6 


| sol Su 


IX; B, 3 Willets: Intestinal Helminthiasis 


TABLE III.—Distribution, Manila versus provinces. 


| 


237 


Manila. Provinces. 
Total. 
Examinations and Filipinos. Chinese. Filipinos. Chinese. 
infections. | 
i] 
Num-| Per |Num-| Per |Num-} Per |Num-| Per |Num-| Per 
ber. | cent. | ber. | cent ber. | cent. | ber. | cent. | ber. | cent. 
eva Eo 
Examined ______-------- , 8,253 |------- IS Eo Peer Sh eee fh 2B Lt ema WERAS, | eit 2 
Positive ees 2,571; 79.0} 360| 30.6 | 2,369} 74.6 121 | 48.2 | 5,421) 69.1 
| { = ————— : =| —————w 
Trichuris ---------- 11,978 60.7| 187) 15.9) 1,474| 466 56) 22.3|3,690| 47.1 
PASCUA =? 2 Lot. 2. | 1,591 | 48.9 188 | 16.0 | 1,355| 42.8 | 77 30.7 | 3,211; 40.9 
Hookworm --_---__--- 619 | 19.0; 12 6.1 1,024] 32.4) 22; 8.8] 1,737) 22.2 
Strongyloides___-_-. i eee 0.9 4 0.3 31 1.0 | a 0. 4. 65 0.8 
Ozyuris Bh ewebtcoted i 24 0.7 2 6.2 31 BU ee [eee ook 57 0.7 
WT eee eee | 13} 0.4 2 0.2 | 40 1.3 2 0.8 57 0.7 
Miscellaneous --_-__- j a 5] 0.4 4 BM RONAN Sees e ast ee 15} 0.2 
Total infections = 4, 254 | 130.8 460 | 39.1 | 3,960 | 125.2 | 158 | 62.9) 8,832 | 112.6 


1914 


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240 The Philippine Journal of Science 1914 


TABLE V.—Summary of Garrison’s helminthic findings. 


Examinations and infections. Number. | Percent. 

Dp é: Xai) 3 (os lament eel ee a Nee cates A ue een la Eee ye) See Neer eS eee er eo 4° 106) }22en 2-86 

| Prichwuriss2-=-—-= | 2, 426 | 59.0 | 
Hookworm’ 22:2. -..2-=-2-5--7. 2, 135 | 52.0 
Asearis: 2.2) 2 253 ee eee 1, 052 26.0 
Strongyloides 182 | 3.0 
Ony aria oe os ee ee ee eee S25; 0.8 

Yio Ty a Me LS a fe MNT 30 | 0.7 | 

Paragonimus 18 | 0.4 | 
Schistosoma. = :_< 3-5-2 ae ee ee ae ae 16 0.4 
Opisthorchis [Clonorchis] ___- ll 0.3 
Higenén lapis 25. Ge i oe ee eee eee ee | 5 | 0.1 


TABLE VI.—Reéxamination of 930" prisoners for intestinal helminthiasis. 


Examinations and infections. Number. | Per cent.| 

Examined ...22< -2225.5 202. su oe 5 ee eee 80 aes cae 
Prichwrte ss: 3-252 ot heh ek, Fe Se a ee 248 26.7 
AS Caria. 22-6 ot oe ie ie Bs eR ee A ee ee 60 6.5 
Hoolkworm \.925 2225 oop os. 2 sees ee ee ee ee 53 5.9 
Onnypurrit’ 2 Soe 535 2 a3 a pe ee ee ee A ee es 41 4.4 
: Sbrivngepbonibe ss os EE Ee he a rn is a ea 8 0.9 
TNE, 22 Paw a oS ae ac a ee es 2 a ee 1 0.1 


“Length of residence in the prison was as follows: Less than one year, 241; from one 
to two years, 150; from two to three years, 356. 


BACILLARY DYSENTERY: THE MOST PREVALENT FORM IN 
MANILA AND ITS TREATMENT 


By W. E. MuSGRAVE and A. G. SISON 


(From the Department of Medicine, University of the Philippines, and the 
Philippine General Hospital) 


The word “dysentery” should not be used in medicine except 
to represent the “clinical complex” of bloody stools, tenesmus, 
etc., for which it was originally employed. In this sense, it 
represents many diseases both pathologically and etiologically. 
In fact, any type of colitis, whether primary or secondary, may 
be expressed clinically as “dysentery.” On the other hand, 
definite pathologic lesions of the colon of specific etiology at one 
time may be associated with “dysentery,” and at other times may 
exist without material disturbance of the bowel evacuations, and 
in still other instances may be associated with constipation. Such 
conditions are so well known that they need not be discussed. 

Scientific nomenclature, at least, should recognize diagnoses 
based upon etiology as far as possible. When the etiology is 
unknown, naming the pathologic condition as a diagnosis is 
acceptable, and only when both etiology and pathology are not 
known is it justifiable to use a “clinical syndrome” as a diagnosis. 

This is particularly true of diseases associated with disturb- 
ances in the evacuation of the bowel contents, whether as “dysen- 
tery,” “diarrhea,” or “constipation.” 

In no other classes of disease are “‘slipshod” methods of diag- 
nosis, which are based upon the obvious clinical symptoms, 
fraught with more danger to the patient; and there are no dis- 
eases where there is less excuse for perpetuating the practice. 

The “bacterial dysenteries’—the subject assigned for this 
discussion—include a large and important group of the colites 
of the large intestine. 

The bacteria concerned are of many species. Some of them 
are rather definite and positive in action, others are influenced 
in their pathogenicity to a material extent by conditions of en- 
vironment and resistance on the part of the host, and still others 
produce definite lesions only under unusual conditions. The 
unusual conditions may consist in intrinsic variations in the 
toxicity of the bacteria, modifications in the resistance of the 
host, or differences in environmental conditions within the intes- 


tine or elsewhere. 
241 


242 The Philippine Journal of Science 1914 


For example, as is well known, staphylococci, streptococci, 
the colon bacilli, and other organisms frequently, if not usually, 
are found in the normal intestine, and it is a fact that under 
certain conditions these organisms may be the responsible agents 
in an acute colitis. Bacillus dysenteriz under ordinary circum- 
stances is found associated with, and is accepted as the cause of, 


one type of acute colitis. Under certain conditions of immunity | 


on the part of the host, decreased virulence of the strain of the 
organism may occur, and under other circumstances which 
probably are not explainable with our present knowledge this 
bacillus may persist for long periods of time in a perfectly 
healthy intestine—the bacillus “carrier.” However, let this 
protected balance be broken in any of its links as by (a) de- 
creased immunity on the part of the host, (b) increased virulence 
or volume of infecting agents, or (c) altered environmental in- 
fluence, and a severe colitis results. 

All of these influences are important considerations in the 
spread of the infection, and in communities where public health 
and personal hygiene are not the most efficient it is largely by 
establishing balanced combinations of these conditions that epi- 
demics are controlled. 


HISTORICAL 


Shiga? described a bacillus occurring in an epidemic of dysen- 
tery in Japan, and concluded that this organism was the cause 
of the disease. Flexner? described a very similar bacillus occur- 
ring in an epidemic of dysentery among American soldiers in 
the Philippine Islands. Strong and Musgrave® isolated a very 
similar organism from cases of acute dysentery in Manila, and 
proved its etiologic relationship to the disease by experiments 
on animals and in one case on man. 

Kruse‘ described a dysentery bacillus similar to Shiga’s as 
being the positive agent in an epidemic of dysentery in Germany. 

Careful laboratory workers very soon began to notice slight 
cultural differences between the various strains of the organisms 
isolated by the different workers, and various classifications of 
the group have been made as a result of studies from the different 
laboratories. 


*Centralbl. f. Bakt. (1898), 23, 599. 

* Bull. Johns Hopkins Hosp. (1900), 11, 281, and Centralbl. f. Bakt. 
(1900), 28, 626. 

* Rep. Surgeon-General U. S. Army (1900), 251. 

* Deutsch. med. Wochenschr. (1900), 26, 637. 


ee 


IX, B, 3 Musgrave and Sison: Bacillary Dysentery 243 


Among the important contributions in regard to this are 
those of Hiss and Russel,®> Martini and Lentz,® Shiga,’ and many 
others. 

During the past decade it has been shown conclusively that the 
dysentery group of organisms is world wide in its distribution, 
that it is the principal cause of epidemic dysentery, “jail” dysen- 
tery, “camp” dysentery, etc., and that various members of the 
group are important causative factors in infantile diarrhoea and 
“‘ileocolitis.”’ 

The types of organisms and the clinical varieties of the infec- 
tions are of importance to all students of tropical medicine, and 
considerable work has been done on the subject in various warm 
countries. 

THE PHILIPPINE ISLANDS 


The influence of these organisms is of special interest to 
practitioners of medicine and students of pathology in this 
country. 

Acute colitis was one of the most serious problems which con- 
- fronted the United States Army during its early days in this 
country. 

The first published work on the subject from the Philippine 
Islands was an article by Flexner and Barker.® 

The authors who represented the Johns Hopkins University 
as a commission had ample material in the epidemic of acute 
dysentery among American soldiers in Manila. They isolated 
and described an organism very closely allied to the one pre- 
viously discovered by Shiga in Japan. They considered the 
bacillus to be the cause of acute dysentery seen in Manila at that 
time. 

Strong and Musgrave® studied over 1,300 cases (with 271 
autopsies) of dysentery, principally among American soldiers, 
and recognized two varieties of the disease: (1) Acute specific 
bacillary dysentery and (2) amcebic dysentery. From the acute 
bacillary form of the disease, they isolated an organism which 
was identical with that of Flexner and Barker and which was 
shown to be very similar to, if not identical with, the original 


5 Med.. News (1903), 82-89. 

*Zeitschr. f. Hyg. (1902), 41, 540. 

*This Journal (1906), 1, 485. 

*Phila. Med. Journ. (1906), 6, 414; also, Bull. Johns Hopkins Hosp. 
(1900), 15, 231. 

*Annual Rep. Surgeon-General U. S. Army for 1900; also Journ. Am. 
Med. Assoc. (1900), 35, 498. 


244 The Philippine Journal of Science 1914 


organism described by Shiga. The authors proved the etiologic 
importance of this organism by experiments on animals and in 
one case on man, and they were the first to isolate the bacillus 
from the lymphatic glands. 

Shiga’® reported extensive comparative studies between 
various strains of the organisms, including the ones from Manila. 
He credited the proof of the etiologic status of the organism to 
Strong and Musgrave. 

Ohno" published an extensive study of all the available strains 
of dysentery bacilli. He elaborated the classification into five 
varieties. 

Bowman? studied an epidemic of infantile dysentery in 
Manila during August, 1907, from private patients selected from 
our service. 

An organism of the typhoid-colon group with certain definite 
distinctions was isolated from all these cases. This organism, 
which Bowman designated as Bacillus “S,”’ appeared to be the 
causative agent in this epidemic. 


In cultural characters, Bacillus “‘S” is smaller and more delicate . 


than B. coli or B. dysenteriz, and it is actively motile. Toxic 
properties and comparative agglutination tests seem to show 
that it is distinct from other organisms of the group. However, 
it is rather closely allied to Morgan No. 1, which was found by 
Phalen and Ashburn in one epidemic of acute dysentery in and 
around Manila. 

The prevalence and influence of the “Y” bacillus of Hiss and 
Russel'® in infantile dysentery and further studied by Hiss™ 
have been studied in the Philippine Islands by Whitmore. These 
organisms are being assigned an important etiologic réle in 
colitis of children in Europe,’® and should be studied partic- 
ularly with reference to the benign dysenteries among children in 
this country. 

Phalen and Kilbourne’® described an epidemic of bacillary 
dysentery in Luzon. 

Bacteriologic studies of 8 cases at Camp McGrath showed 
Morgan’s No. 1 bacillus?” in 5 cases, but no dysentery organ- 


* This Journal (1906), 1, 485. 

“ Tbid. (1906), 1, 951. 

™ Ibid., Sec. B (1908), 3, 31. 

* N.Y. Med. News (1908), 82, 289. 

* Journ. Med. Research (1904), n. s. 8, 12. 

* Arch. f. Kinderheilk. (1918), 60-61, 35, 689. 

* Milit. Surgeon (1910), 26, 433. 

* Brit. Med. Journ. (1906), 1, 908; (1907), 2, 16. 


— 


1X, B, 8 Musgrave and Sison: Bacillary Dysentery 9A5 


ism was obtained in the other 3 cases. Two dysentery cases 
out of 4 from Camp Eldridge, Laguna, yielded Morgan’s No. 1 
bacillus. Of 2 cases examined at Fort William McKinley, Mor- 
gan’s No. 1 bacillus was isolated from one; the other case was 
subsequently diagnosed as ameebic. _— 

Of 3 cases studied in Manila, one was negative, the Shiga 
type of B. dysenteriz was isolated from one, and the Flexner 
type from the third. 

These authors recognize two main groups of dysentery-pro- 
ducing bacilli, the Shiga type including Kruse’s organism and the 
Flexner type which includes those of Strong and Musgrave and 
of Gray and Duval. Other organisms which had been considered 
as probable causative agents at that time were Bowman’s 
Bacillus “S.” 

They conclude that: 


Bacillary dysentery is endemic in the Philippine Islands at all times, 
occasionally assuming epidemic proportions over more or less wide areas. 
The disease is due to at least three types of dysentery organisms, the well- 
known Shiga and Flexner types and the less known organism of Morgan. 
In the region south of Laguna de Bay the Morgan type alone was encoun- 
tered, and this organism appears to be the predominating cause of the recent 
epidemic. In and about Manila, where the cases were more scattered and 
scarcely assumed epidemic proportions, organisms of the Shiga and Flexner 
types were found. * * * 

It does not appear that the disease caused by one type of dysentery 
bacillus differs from that caused by another in its clinical course, its 
pathology, or its gravity, and the treatment is the same for all. The 
identification of the bacillus type, then, in any given case, does not seem 
of importance. The diagnosis of this variety of dysentery must still depend 
upon the clinical course of the disease and the exclusion of amceba by 
microscopical examination. 


Whitmore® studied an epidemic of bacillary dysentery in the 
Philippine Islands during the summer of 1909. Bacilli, resem- 
bling those found in dysentery, were isolated from the stools of 
a number of the patients affected, and the author made a com- 
parative study of the various strains of the dysentery bacilli. 
He concluded that they were principally of the Shiga-Kruse 
type, with a third class less frequently found which includes 
the “Y” bacillus of Hiss. The Shiga-Kruse type of organism 
was the only one found in cases studied from the provinces, but 
the Flexner type was encountered in cases in Manila. The 
author makes no attempt to associate the type of the bacillus 
present with the severity or the clinical variety of the disease. 


“This Journal, Sec. B (1911), 6, 215. 


246 The Philippine Journal of Science - . 1914 


Chamberlain, Vedder, and Barber,’® constituting the United 
States Army board for the study of tropical diseases as they 
exist in the Philippine Islands, reported an unusually severe 
epidemic of acute bacillary dysentery raging around the town 
of Ormoc, Philippine Islands, during the early part of 1912. 
The disease was most prevalent in children and babies, and 
the mortality was extremely high. The observers arrived on 
the scene only toward the end of the epidemic, and were unable 
definitely to isolate the dysentery bacillus. However, they are 
of the opinion that the epidemic was due to some variety of 
the dysentery bacillus. From one case they isolated an organism 
corresponding in all essential details, except that of mortality, 
with Bowman’s Bacillus “S.” 

Barber and Gomez ”° studied an epidemic of acute dysentery 
at Baguio. There were 23 cases in all; 9 Americans with 2 
deaths; 1 Filipino, no death; 11 Igorots with 8 deaths; and 
2 of other nationalities with no deaths. They isolated a dysen- 
tery bacillus from 14 out of the 23 cases studied. Of the 14 
strains of the organism, 10 were of the Shiga type and 4 of 
the Flexner type. All of the native Igorots of the mountains 
showed the Shiga type of organism. In all cases where it was 
possible to trace the source of infection it was found that the 
disease in Igorots or in foreigners was contracted at one of 
the railroad camps. Examination of the pail system of this 
camp showed that there was a large amount of bloody flux 
among the workmen. Laborers in this camp included all nation- 
alities, and were received from all parts of the Philippine Islands 
and from Japan. 

Ashburn and Vedder 7! studied an epidemic of acute dysentery 
prevailing among United States soldiers at Camp John Hay near 
Baguio. The authors stated that during the year 1911 and part 
of 1912 there were 81 cases of acute intestinal trouble among 
the troops in that camp. The clinical symptoms were those 
usually referred to as “mountain diarrhoea” characterized by 
diarrhea with mucus and blood in the stools and by slight fever 
lasting from two days to a week. From one of these patients 
the authors isolated a dysentery bacillus of the Flexner type. 

During 1912 an epidemic of acute intestinal flux among the 
troops of one company was studied. The symptoms were mild, 
consisting of diarrhceaa—some blood and mucus—the disease 


» Milit. Surgeon (1912), 30, 318. 
»” Bull. Manila Med. Soc. (1912), 4, 138. 
™ Ibid. (1912), 4, 139. 


1X) B,-8 Musgrave and Sison: Bacillary Dysentery QAT 


lasting for about a week in each case. By agglutination tests, 
the authors arrived at the conclusion that the epidemic was 
due to bacilli of the Flexner type. 

During the fiscal year 1912-1913, excluding cases of ameebic 
and other extraneous types, there were treated in our service in 
the Philippine General Hospital 266 cases of acute colitis. 

Distributed according to ages, these cases were as follows: 


| Age. Cases. Age. Cases. 
PAS VEATE ee eons se cen OU ean tO AO wears osteo ee Se ee 20 


PEI OVERTS Hse oe dor AO aby Kearse eye ee 4 
ROOM OIYGBYS) 2-25 02-25 cosesak ||| Aor tO DO Vea re)... a Ane ne otk 4 
HENEGAU VY CAY Al se 2 = pee ne eS Sbit|) DO tobS yearaies 2 ean ee ee 2 


ITO GG: ii 4b il 55 bo 6Oigenre 222 o's he Stet COE oF 1 
Mame OU VEAER (20525-2250. 98 eo 37 PAgR otal on Peas Sie, 266 
SUnRO TO Vea ne 50) oe een 28 | 


There were 191 males and 75 females. 

The total mortality in the 266 cases was 49, or 18.4 per cent. 
Autopsy was performed on 29 of the cases. The mortality 
among males was 34, or 17.8 per cent, and among females 15, 
or 20 per cent. 

The mortality according to age is shown in the following 
table: 


| Age. Cases. |Per cent. Age. Cases. |Per = 


Nita'G Vearse— =~ 26. .2==..- 24 9.0 || 30 to 35 years ._....____._--. 2 0.7 
‘ato 10 years... 3...-:-2=. 5 19)}|'35,to 40 years _..<--=-=-2- 5. 2 0.7 
NOjtontb years. .t264. 22-5 0 0 40 to 45 years ___________-__- 1 0.4 
i5 to 20'yeara -.-._....=.... 2 0.7 || 45 to 50 years ______________- 3 it 
20:to 25 years _.....-.--..-- 5 1.9 || 50 to 55 years ___________-_-- 0 0 

| 2opo'S0years,.22—....--_ ... 4 1.5 || 55,to 60 years _........._____ 1 0.4 


In considering the mortality statistics in these cases, it must 
be remembered that they were all hospital cases and consequently 
represented patients with severe clinical symptoms. 

The mortality from this disease in other hospitals is given by 
various authors as follows: 

Japan, 16.5 to 30.2 per cent. Ceylon during 1903, 28.7 per 
cent. British New Guinea in 1902-3 at 22.8 and 26.6 per cent, 
respectively. In Singapore where the dysentery is considered 
mild, the mortality in the hospitals in 1902 was not less than 
25.4 per cent, and in the neighboring state of Selangor it reached 
34 per cent. In Trinidad it stood at 30.7 per cent, in German 

126334——4 


PAS The Philippine Journal of Science 1914 


New Guinea at 33 per cent, and in Hongkong in 1902 it reached 
37.3 per cent. 
ASSOCIATED DISEASES 


In 61.6 per cent of the cases, the colitis was associated with 
other diseases, and the mortality in such double diseases was 
57.1 per cent. 

The principal associated diseases with their incidence is shown 
in the following table: 


| Associated disease. Cases. Associated disease. Cases. 
Phthipis2:-2c- 2c. eemesenee ce ees 18) || VOSyeridsis’= see een eae 2 
Duodenal'uleer —2-22=~ <== = eee ||) ;Atmocebinsieste ss ae Se ee 10 
Aiscariasisis:=:2e8¢ o- seme. ord 48 || Strongyloidiasis -___.---_.-_.---------|. 1 
Trichurigeipeo-se ce cae eee ee 57 || Tubercular intestine________________.- 2 
Ankylostomiasis --_-_-.-_--------.---- S11) Malaria: 3. = eerie en sees eS 
| Monadiasis: .2.25.22552 3) ee eee ll 


COMPLICATIONS 


In 30.1 per cent of the cases other conditions believed to be 
complications of the bacillary infection were encountered, and 
the mortality in the presence of complications was 59.1 per cent. 

The complications noted, with their frequency, are as follows: 


Complication. _ Cases. | Percent. Complication. Cases. | Percent. 
Maniaacute 6-22 sc -_ 84 2 0.7 || Rheumatic fever --_-_-_.--- 2 0.7 
Nephritis, acute___-_.----- 37 13.9 |} Lymphadenitis -.-__.-----_- 8 3.0 
Bronchopneumonia-.----_-_-. 14 bez |) Gisophagitis 22-02-5222 422 1 0.4 
Bronchitise cacs sean conan 9 3.4 || Splenitis, acute_.----.--_--- ; 1 0.4 
Acute cardiac dilatation -- 5 179) Meninortiss..22.-c 225 22o5 0° 2 5 1.9 
Gastroenteritis-___--_------ 6 2.80)\ SLY Grobnorax: 22. 5- 85-5 7ooa6 2 0.7 
Neuritis, multiple -________ 2 0.7 || Fibrinous pleurisy_---_----- 4 1.5 
PROPMONM pone a+ eee ees 6 2.3 || Hydrocephalus -__-.._------ 2 0.7 
iProlapsusiani 2. -------- = 2 O27) |WReritonitif==-.2s--s-2.ee cae 6 2.2 
Hemorrhoids --_---._------ 4 1.5 || Pneumonia, lobar -----__--_- 2 0.7 


These complications, while not all due to the influence of B. 
dysenteriz, must nevertheless somewhat elaborate our previous 
conception of the extensive harm which may result from this 
organism. 

For example, Strong and Musgrave, out of a total of 271 
autopsies performed in 1899, encountered 111 cases of colitis. Of 
these, 21 were classified as acute specific bacillary dysentery, 
11 as subacute, and 79 as ameebic dysentery. The complications, 


| 


——_— 


IX, B, 3 Musgrave and Sison: Bacillary Dysentery 249 


associated diseases, and special anatomic findings in the acute 
bacillary type of the disease are recorded as follows: 


Malaria fever 

Chronic gastric catarrh 

Bronchopneumonia 

Acute pleurisy 

Congestion and cdema of lungs 

Cloudy liver 

Cloudy kidneys 

Fatty liver 

Fatty kidneys 

Enlarged spleen 

More recent writers have included a number of other complica- 

tions of these infections to such an extent that the dysentery 
toxin must be classed with that of other severe infections in its 
action on organs and parts of the body distant from the seat 
of lesion. 


or rR OOO RE wD bd 


TREATMENT 


In spite of our knowledge of the etiology of this disease, we 
have not as yet a treatment that might be termed specific. This 
is explained by our knowledge that different types of micro- 
organisms are causative factors of the disease, and this fact 
together with the different degrees of virulence of the infecting 
microorganisms in part explains the disparity of opinion regard- 
ing the treatment of bacillary dysentery. The treatment recom- 
mended as effective by some clinicians has been a failure in the 
hands of others and vice versa. Many physicians who have had 
experience in the treatment of bacillary dysentery are in favor 
of some special drug or of a manner of treatment that has given 
them the best percentage of cures. 

There is one phase of the treatment, however, where all 
the writers are in accord; that is, the prophylactic treatment. 
With our knowledge of the etiology of the disease, it is not 
difficult to observe the measures that will prevent the spread 
of the disease. Each individual with dysentery must be con- 
sidered as a focus of infection, and his isolation is necessary. 
The stools must be properly disposed of, and general hygienic 
rules must be observed. Overcrowding, uncleanliness, over exer- 
tion, fatigue, and the use of injudicious diet are some of the 
predisposing factors that ought to be avoided, especially during 
dysentery epidemics. All materials that have come in contact 
with dysenteric patients and the hands of those who attend them 
should be disinfected. Flies are the main disseminators of the 
disease, and must be excluded from contact with the patient 
as well as with food supply. 


250 The Philippine Journal of Science 1914 


We have had several infections that have occurred in the 
Philippine General Hospital during the years of 1911-12 through 
the agency of flies, as we had a pest of these insects at the time 
we were taking care of patients suffering from bacillary 
dysentery. So also the epidemic of dysentery in Baguio during 
the summer of 1912 was evidently spread by flies, as is proved 
by the article of Banks read before the Manila Medical Society 
in the same year, entitled The Baguio fly campaign. 


CURATIVE TREATMENT 


Our routine treatment in the Philippine General Hospital which 
has given the most satisfactory results consists in the following: 

First, absolute rest to save the strength of the patient and 
to prevent the involvement of the larger segment of the intestine. 

Secondly, the early administration of some mild laxative, pre- 
ferably sodium sulphate or magnesium sulphate, preceded by 
fractional doses of calomel in order to diminish the presence 
of infecting material in the gastrointestinal tract, as well as to 
get rid of some irritating material that might be present there. 
The administration of simaruba officinalis combined with some 
opiate is highly recommended, for it has given us the most 
satisfactory result in comparison with the use of other drugs. 
As an adjuvant to this treatment, the judicious use of normal 
salt solution as an enema, or given in the form of the drop 
method per rectum in the amount of 1 liter once a day, is some- 
times very beneficial. 

When the acute stage of the disease has subsided, enemas 
of hydrogen peroxide in a weak solution (about 25 cubic centi- 
meters in 500 cubic centimeters of water) once a day are a great 
help toward prompt recovery. 

The use of ipecac, although strongly recommended by some 
writers, has not given us universal satisfaction. While some 
patients are benefited by this drug, there are a great many 
cases in which the use of ipecac becomes another sort of torment 
to the sufferer. There are many persons who abhor this drug 
to such an extent that even the smallest amount of it, though 
combined with opium, is apt to produce in them marked 
emesis. We have seen cases that are not even able to tolerate 
Dover’s powder given in almost homeopathic doses. This is the 
greatest drawback to ipecac, and the deémetinized preparation 
is almost useless as the active principle of the drug, the emetine, 
is the one that has some therapeutic action in the treatment 
of dysentery. The use of astringents and the so-called gastro- 
intestinal antiseptics we have given up as unsatisfactory. Al- 


1X, B, 3 Musgrave and Sison: Bacillary Dysentery 251 


though there is a small percentage of patients who recover 
under this treatment, we should remember that there are cases 
that get well even without any medical treatment, and we con- 
sider it problematical whether or not the usual astringents and 
gastrointestinal antiseptics are really beneficial in the treatment 
of this disease. We must keep in mind the fact that most of the 
astringents and so-called intestinal antiseptics, such as, bismuth, 
tannic acid preparations, salol, beta-naphthol, benzo-naphthol, 
benzoic acid, and others, have an irritating action upon the 
stomach and intestines especially if they are given in large 
amounts and over a considerable period of time. 

The use of the ice bag over the abdomen is a great help in 
diminishing the abdominal pain, and hot turpentine stupes fre- 
quently are useful for the same purpose. 

The essential part of the treatment, however, is dietetic. 
During the first twenty-four hours of the acute stage of the 
disease food must be withdrawn. Pieces of cracked ice may 
be given to allay thirst. At the end of twenty-four hours, we 
allow the patient albumen water or rice or barley water and 
later skimmed or peptonized milk. When improvement has he- 
gun, milk, broth, beef juice, and orange juice may be given. 
The mouth must be frequently cleansed with an antiseptic mouth 
wash to prevent the frequent complications of parotitis and 
gastritis. 

The serum treatment, first recommended by Shiga in 1898, has 
both advantages and disadvantages. If the variety identification 
of the bacillus which is the cause of the infection can be carried 
out with readiness, as well as with accuracy, this scientific treat- 
ment ought to yield a greater percentage of cures than usually 
is obtained. For practical purposes, however, especially in those 
cases that have to be treated in places where the means of 
identifying the infecting microdrganism are not available, it is a 
failure in most instances. The sera of patients suffering from 
one form of bacillary dysentery usually will not agglutinate 
other varieties of Bacillus dysenteriz. 

It is possible that Flexner’s polyvalent serum might be used 
for any acute bacillary dysentery. However, what we have 
already mentioned in the discussion of the etiology of dysentery, 
in regard to the etiologic importance of other microdrganisms, 
such as streptococci, staphylococci, colon bacillus, and others con- 
cerned in the production of colitis under certain circumstances, 
will make the use of even a polyvalent serum unsatisfactory in 
many instances. 


ate EE Te URES, Hae TO ogee 


5 BETTS bod) ¢ yy thet RA ake PR eae fa ae ie 
RAD So eae ES at ek | MERE tae) 
P aioe Pte teat ts Ling isa: Be ae 
WRT toa) Mean oe tai octal deg See 
Ae DARRY ah i oi rale tds hea ey 
vif Breet are 62 ans re st lal th ai 
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rt PORES Pe nero te yt Bernt eh 
OR ie) Rare mena te teas Aiea yhouipaeh a 
eR La) oie wertider ot bity rather : 


D tr haetial | 


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amity Clo S54, Op ae GN och Pauper ut a 
PRS depend iy hee ee TA Re ried Yel Pian vis 19a 
a tah ae i ee ee fiirsebibettis Fear tay 
tne OA che Tpeeaattiaten mi ad Y 
trek! erated Tx) Reva We 
Behewiols > 2 Sapes ‘Ki ‘Savard 
(abies ender ny Wit rttalg | 
‘riteiien sep Siig leis ade me | 
MRT Ns Ta ey iain 24% 
sf Ret RT Tey ee, ‘sas a e inte ay 
PAG) NN) ei) ae ORT ee tra an CA ea ie 
Cinnien musts oY 2h Take Sead bla aw a 
pat Peay iat ‘ity peasant peg ate Maer, 79% eae 
; ‘ty ood vale sh fone of ai Shah ody 
' phatiadioey ie -# Hh De Me TAY Louris wera it 4 
Mlerit cis Tee AR St ae Pe aie SHES Renee ath jada a: 
Noi POPOL thay yg Vat res Parte ne Ba 
; sit eta! YAW ts a bate artis 
‘ 1 kai ith (oe lash yetogy: Petals Bile ‘dial *: 
el eieb aR 16th Paes iy OM we Us oe rarely tsdiltanill 4 it 
MEA tes hs a ea ee eee 
Hie oe hor gedit y ese 
i a ets Watéy Nipiey esk Se i 
milfs rota SAH \ 
nae sie heir iNa iiporie el - i 4 


hile Rae BS 


a 


WIDAL REACTIONS AMONG HEALTHY ADULT FILIPINOS 


By Davip G. WILLETS 
(From the Biological Laboratory, Bureau of Science, Manila, P. I.) 


This work was undertaken for the purpose of determining 
(1) the percentage of healthy adult Filipinos indicating previous 
or present infection with Bacillus typhosus by the presence of 
a positive Widal reaction and (2) the percentage of typhoid 
carriers among this class of individuals. The second phase of 
the investigation was to have been conducted by Dr. J. A. Johns- 
ton of this laboratory, but this work was unfortunately inter- 
rupted.: The results of the Widal tests alone, however, indicate 
the advisability of placing them on record. These tests were 
made on the blood of 300 apparently healthy, young, adult Fili- 
pino employees (nurses, etc.) of the Philippine General Hospital. 
In making the tests, serum only was used. The dilutions and 
time limits are given in the following table. The microscopic 
method was used, because it was desired to apply to these cases 
the same routine test as to suspected typhoid cases. In the midst 
of the investigation, the typhoid culture was examined morpho- 
logically and culturally and found to be quite pure. It may also 
be stated that from 15 to 30 tests were made at a time and that 
these were checked against reactions on the blood of known cases 
of clinical typhoid fever. It will be noted that control reactions 
were performed in a number of instances. These were made 
about ten days after the original tests. 

After cleaning a portion of a finger or of an ear with 95 per 
cent alcohol and drying with sterile cotton, a small quantity 
of blood was collected in a capillary pipette. The smaller end 
of the pipette was closed in a Bunsen flame, and the larger end 
with sterile cotton. The specimen was then centrifuged until 
the supernatant serum was quite free from blood corpuscles, 
whereupon it was placed in a refrigerator for a period of from 
a few hours to one day. The serum was drawn off by means 
of a fine capillary pipette and placed in a watch glass. It 
was diluted 1 to 20 by use of a pipette for counting leucocytes. 


*In the few cases in which cultures of the feces and urine were made, 
B. typhosus was isolated in one instance; this case had given a definitely 
positive Widal reaction. 

253 


254 The Philippine Journal of Science 1914 


Equal portions of it and a suspension of typhoid organisms were 
placed on a cover glass by means of platinum loops of equal size 
and thoroughly mixed. This constituted the hanging drop used 
in connection with a depression slide. All glassware was clean 
but not sterile. In diluting the serum, a separate portion of 
normal saline solution was used for each specimen. ‘The sus- 
pension was prepared from an eighteen to twenty-four hour old 
acid agar slant subculture with normal saline. The hanging 
drop was carefully ringed with vaseline. Microscopic obser- 
vation of the hanging drop was made every few minutes. The 
1 to 80 dilution was prepared by taking equal portions of a 
1 to 40 dilution of serum and bacterial suspension. 
Noteworthy agglutination and loss of motility were present in 
49, or 16.1 per cent, of the cases as indicated in Table I. This 
was of such a degree as to be considered definitely positive in 
12, or 4 per cent, of the series of cases. The positive cases are 
numbers 1, 2, 9, 18, 20, 22, 28, 25, 26, 29, 40, and 49 in the table. 


TABLE I.—Reactions in 49 of the 800 cases examined. 


[Clumping and loss of motility absent (A), complete (C), and partial (P).] 


Original reactions. 
Check reactions, 


Dey Vie athe dilution 1 to 40. 
No. Dilution 1 to 40. Dilution 1 to 80. 
Minutes.} Result. |Minutes.| Result. |Minutes.| Result. 

Se Se Se RE Ne oe eee ae 40 , 0 eae ee erento eee | See eme eel 

VG Se ASA Se Uae Fe Bet) eps se 30 Gees eee cae eae er MC ep eee | Ne 

Bee. sees 5. Re ARE LD See ee Ree 55 On emeriare aee alee EROS E Eee teats ee A 

7S Ree Pee ee ss Se eee ees A 60 Ve 90 Phy rile os Boreas ill = os, cee es 
(7 oe ee Se ec eens Pe oe ee 60 P 90 ip 90 P 

Gea IN ee hed Ce Ss oe 60 P 90 Per) Aeceeceeses| Goce 
/ (ey eee SE ee Ss aes eee 60 Pe 90 iz 90 P 
ilps 2 tle he os et 60 P 90 ip 60 Pp 

ee ce Se ae 30 Cc 60 Co ee cae wad | Se 
a (Jyh a ape hae ee Ea 60 P 90 A 90 A 
rE | eee Oe Ss ee ee 8 ee eR | 60 122 90 y 90 Js 
Use Rect Deed Aen aod 50 Cc 90 A 90 cy 
1 ee Sea Saree ae ores eee ere 20 Cc 25 Cc 60 P 
TY Reta ule ee Se Se ae Se ei Soe 50 Cc 90 A 90 P 
Ws 4. RAK ~ SEE ee 60 iP 90 A 60 P 
MGeo cet. A en ee eee ee le 60 P 90 A 90 P 
DT oa 8 eee ee ae ee ae ea 60 a 90 A 90 1 
18 fos coe Be Je ea eee 60 ib $0 P 60 P 
19.5 [Mw See een 60 P 90 P 90 P 
60 P 
| a ES Rea ee ee aS | 45 Cc 60 Cc | 90 Cc 
QU ae ce hl Se oe 60 P 90 P 60 P 
BO os Ee eg Aer 35 Cc 90 Cc 25 Cc 
V2: eee een apre Seen, emma k BIL e 50 Cc 60 Cc 45 Cc 


See ye 


IX, B, 8 Willets: Widal Reactions Among Filipinos 255 


TABLE I.—Reactions in 49 of the 300 cases examined—Continued. 


Original reactions. 
Check reactions, 
dilution 1 to 40. 
No. 


Dilution 1 to 40. Dilution 1 to 80. 


Result. |Minutes.| Result. 


| Minutes.| Result. | Minutes. 
OSE eT ES ea eee 60 P 90 A 60 A 
i seat ae stl he Eo eal 45] C 90} C 2051. G 
To ee eee Sn eee Se 40 Cc 60 Ct ER cee eae 
pypueenins ecco Beet Sie, a lew 60 P 90 A 90 P 
Fi nt Oe a 2 en ee 60 P 90 P 90 A 
2 wrens Aes ie ee eee 30 Cc 60 c 30 Cc 
cee ese ah 60 P 90 P 90 P 
Pine eee ge 60 P 90 P 60 P 
Foi oa ee ee ee 60 P 90 P 60 A 
Te tee tl 60| P 90 A 60 P 
Pameererees rile UN te. 60 Cc 90 Cc | Bi SE ea ee 
peeeeeee PIT MLS AG 2. 60 Cc 90 Goa VL eee ed 
Te eee ee ee ee 60 Cc 90 Bevr ale eeesoaes fee i 
Reet vee 60 P 90 A Hpeyer. sR ee 
5 ae ee el Peart at eed 60 & 90 pe eles Saree cate Se 
nae lien "tyres: Berth 60} P Sper ph) ath ortels ccee en 
£5, |g 1G SN CY pues oe hk ee 
60 P 90 YM ie A lee fs di Rae 
60 P 90 AEF te RAL: RiGee ARE 
60 P 90 IAG joes. eee Ev 
60 P 90 2 oe et | nee Pa Pe 
60 P 90 id set a Dg 
60 Cc 90 Cc 
60 Cc 90 Cc 
60 Cc 90 P 
30 Cc 30 Cc 


In order to determine the probable significance of the complete 
reactions, the cases diagnosed as typhoid fever in the hospital 
during the year were investigated. It was found that the clinical 
diagnosis was verified by positive blood cultures in 8 cases; by 
anatomical lesions at autopsy in 9 cases; and by both positive 
blood cultures and anatomical lesions at autopsy in 5 cases. 
Widal reactions, performed in a manner similar to those upon 
the healthy adults, excepting that the serum was quite fresh, 
the dilution 1 to 40 or 1 to 50, and the time limit forty or fifty 
minutes, were positive in 20, or 90.9 per cent, of these 22 cases 
as shown in Table II. 


256 The Philippine Journal of Science 1914 


TABLE II.—Fesults of Widal reactions in 22 cases of clinical typhoid fever, 
verified by positive blood cultures, by anatomical lesions, or by both. 


lap Ana Ana: a | 
No. | culture, | fomical |, dction No. culture, | tomical | penetion, 

a aphh elpMgtstcil Ste eee eet + De aa eee kee Orn | eee ede + + 

PAM fet Sa lt NA, + + — (Ge EA ee ae ease = rabies |S ee 2 Eta 

5 ee Res Saayeteie ss) be Sane = 3 0 CEES Se eee 8 + + + 

yh ee ae Balsa SEES ar SP GRR as ee ere SF + + 

1) ee eee ee SSPE Se | eee nee + EG ee ae eer a oe |e ae ee + oF 

Of23 2,2 es Feo en Se = Ly (= apa ee Nt | ai ee + 

Th. Boek Fae i ee + + 18) oo 22 ee a ae + = 

St. 52 ese eles eee + + hCG el Sw eae Soo Wey | eae SI + 

OF eee ee come + ot 20s eee = + a 

108.4 eee oe mens + + Oh Saar e aa eee =— + 
E me allots Abel Bi cineeaboas ek coat 2, 


Only one Widal test was made upon each of the two negative 
cases (numbers 2 and 18 of the table). In the first one, the 
test was made on the fifth day of the disease; in the second 
case, on the eleventh day. Both patients were adults. It is 
highly probable that a positive reaction would have been obtained 
in each of these cases if additional tests had been made. 

In view of the foregoing facts, it seems reasonable to believe 
that the complete reactions among the healthy adults indicated 
an antecedent or a coincident infection with Bacillus typhosus. 

The significance of the partial reactions is problematical. 
Since it is known that the agglutinins which make the Widal 
reaction possible disappear gradually from the blood, some of 
these reactions may also have indicated an antecedent or a coin- 
cident infection with Bacillus typhosus. However, the serum 
used was not inactivated, hence normal agglutinins may have 
played an important role in the production of the partial 
reactions. Furthermore, the possibility of infections with 
organisms closely allied to B. typhosus must also be considered, 
for it is commonly known that the agglutinins produced by these 
bacteria give a positive reaction in low dilutions with Bacillus 
typhosus. 

Chamberlain? reported positive Widal reactions (dilution, 1 
to 50; time, 1 hour) in 9, or 2.9 per cent, of 307 healthy adult 
Filipinos. In the same paper statistics are submitted which 
conclusively prove typhoid fever to be a widely distributed and 
common disease in the Philippines. The records of the Phil- 
ippine General Hospital for the period July 1, 1911, to June 30, 


* This Journal, Sec. B (1911), 6, 299. 


| 
4 
| 
| 


Ix, B, 8 Willets: Widal Reactions Among Filipinos 257 


1913, indicate that the disease is on the increase. In the fiscal 
year 1911, 22, or 1.46 per cent, of 1,509; in 1912, 68, or 1.96 per 
cent, of 3,469; and in 1913, 108, or 3.62 per cent, of 2,985 medical 
cases were diagnosed as typhoid fever. The annual reports of 
the Bureau of Health for 1911 and 1913 likewise tend to prove 
the increased incidence of typhoid or, at least, its increased 
recognition. In a general way, the increased percentage (4 
against 2.9) of positive Widal reactions in our series of healthy 
individuals as compared with that obtained by Chamberlain in 
1910 may be said also to indicate its increased prevalence. 

Table III gives the results of Widal reactions performed upon 
25 employees of the hospital before and two weeks after anti- 
typhoid treatment. If the presence of a positive Widal reaction 
after treatment is an index of protection against typhoid fever, 
it appears that the treatment was effective in 21, or 84.0 per 
cent, of these cases. In this connection it is to be noted that 3 
of the 21 cases were positive before treatment. 


TABLE III.—Widal reactions in 25 employees before, and two weeks after, 
antityphoid treatment. (Serum diluted 1 to 40.) 


Before treatment. | After treatment. Before treatment. | After treatment. 
Nit Smee cases liaa ate ST haere INOS 
Minutes.| Result. |Minutes.| Result. Minutes.| Result. |Minutes.| Result. 


| anes 60 = 60 = 14 os 60 = 30 te 
2a ___ 40 ie 60 So | 55 + 30 ae 
Geese 30 abe 30 aha 16.2522: 60 = 30 oh 
a 60 = 60 = py ee oe 60 = 60 = 
pee 60 = 30 ot ih See ees 60 = 30 5 
Oss —8 60 = 30 3" 1952 eee 60 = 45 + 
(eae 60 = 60 ae CONS Res 60 = 45 + 
LoS 60 — 60 SF jiede se 60 = 45 Se 
1 eee 60 = 30 + he 60 = 40 + 
f0e=—— | 60 — 35 ot pena. 60 = 50 ot 
11> _ 60 = 40 + 60 = 60 =F 
if eaees 60 = DOF a enn 60 = 45 AP 
Use eres 60 = 60 = 


8 Two injections only. > One injection only. 


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uintotiee emobtauen taht hy ting weld uatithg LLY wa 
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views tabi! atertiai Mg cf ie seer DG wath st incerteoe bier 
fev nk dueciepae Nik tedese hi Heeneh: we ph Jthanrters 
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EXPERIMENTS ON THE CULTIVATION OF RINDERPEST VIRUS 
AS DESCRIBED BY BALDREY 


By WILLIAM HUTCHINS BOYNTON 
(From the Veterinary Division, Bureau of Agriculture, Manila, P. I.) 


Baldrey + states that he was able to produce material highly 
toxic for cattle by inoculating broth culture medium with virulent 
blood and allowing it to incubate at 37° C. for twenty-four hours. 

Baldrey conducted his work in accordance with the presump- 
tion that rinderpest serum is an antitoxin and that by injecting 
the toxin obtained by cultural methods, in place of virulent. 
blood, an antitoxic serum could be obtained. In the preliminary 
work, defibrinated virulent blood was inoculated into ordinary 
broth in flasks of 500 cubic centimeters’ capacity in the propor- 
tion of 50 cubic centimeters of blood to 950 cubic centimeters 
of broth. The medium was incubated at 37° C. for twenty-four 
hours. Six immunized animals were injected subcutaneously 
with this, in doses varying from 300 to 700 cubic centimeters. 
Four of these died in from twenty to twenty-four hours, and the 
remaining two were destroyed as a result of the extensive gan- 
grene set up at the seat of inoculation. The post-mortem exam- 
inations revealed evidence of acute toxemia. 

Three animals formerly used for serum production were 
employed to test the possibility of hyperimmunizing animals 
with culture medium and virulent blood after incubation. Mar- 
tin’s broth was employed, but the relative amounts of culture 
medium and defibrinated virulent blood remained as in the first 
experiment. The immunizing properties of the sera produced 
by this method were tested in comparison with other sera. 

Baldrey draws the following conclusions from his experi- 
ments: 

1. That Anti-Rinderpest Serum can be prepared by the inoculation of 
virulent blood diluted with broth. 

2. It appears possible that an active toxin is produced and excreted into 
the Broth by the Rinderpest organisms contained in the virulent Blood and 
by this means the results recorded are obtained. 

3. This material or probable toxin is rapidly excreted and so active that 
it appears to quickly inhibit any further growth of the Rinderpest organism, 
destroying its virulence and finally killing it. 


* Journ. Trop. Vet. Sci. (1911), 6, 251. 
259 


260 The Philippine Journal of Science 1914 


4. The substance so obtained is very much more active than that obtained 
in virulent Blood, so much so that it cannot be given subcutaneously with 
safety on account of the extreme inflammatory condition it sets up. 

5. To use it as an hyper-immunizing agent, doses are better given intra- 
venously and gradually increased so that the “salting” is spread over a 
period of some 2 months or more. 

6. The immune serum so obtained is powerful, but 15-20 per cent. weaker 
than that made by massive injections of virulent blood. 

7. The method is an eminently practical one and shows a great financial 
saving. 

Experiments were performed by me with the view of determin- 
ing whether or not it is possible to obtain the toxin of amie 
by cultural methods, as claimed by Baldrey. 

Both cattle and carabaos were used. A few were immune 
to rinderpest, but the majority were susceptible. 

Experiment 1.—This experiment was performed to deter- 
mine the action of Martin’s broth alone upon an animal. Since 
this medium is highly peptonized, it was thought that possibly 
it might have some effect upon an animal when given in large 
quantities. 

The animal used was Batanes bull 3140, which had previously 
been immunized to rinderpest and had recovered thirty-eight days 
prior to this experiment. On October 4, 1911, this animal was 
injected with 700 cubic centimeters of Martin’s broth, neutral 
in reaction to litmus paper. One hundred cubic centimeters 
were administered intravenously, and 600 cubic centimeters, 
subcutaneously. 

This animal suffered no apparent ill effects from the injection 
of the culture medium. It ate heartily, and was thrifty in ap- 
pearance throughout the experiment. There occurred a slight 
rise in temperature during the first thirty-six hours following 
the injection, which may have been due to the peptone content 
of the broth. 

The result obtained in this experiment would lead one to infer 
that the broth used in the following experiments, by itself, has 
practically no effect upon the animals. 

Experiment 2.—This experiment was designed to duplicate 
the first experiment described in Baldrey’s paper, the only differ- 
ence being the use of Martin’s broth instead of ordinary broth. 

The animal used was Batanes bull 3146, which had previously 
been immunized to rinderpest and had recovered forty days prior 
to this experiment. On October 7, 1911, this animal received by 
subcutaneous injection 500 cubic centimeters of Martin’s broth, 
which had been inoculated with 25 cubic centimeters of virulent 
blood from bull 3217 and kept at 37° C. for twenty-four hours. 


1X, B, 3 Boynton: Cultivation of Rinderpest Virus 961 


This animal showed no ill effects from the injection, and its 
temperature was normal throughout the experiment. 

The results obtained do not coincide with those obtained by 
Baldrey. 

The following experiments were performed with animals 
which had not previously been immunized to rinderpest. The 
object was to determine if possible whether the toxin of rinder- 
pest, if present in the inoculated media, would have any effect 
upon susceptible cattle and also to ascertain if the virus re- 
mained alive after being incubated for twenty-four hours at 
at? CG, 

Experiment 8.—The animal used was Batanes bull 3222, 
susceptible to rinderpest. On October 12, 1911, at 2.30 p. m., 
this animal received by subcutaneous injection 1,000 cubic centi- 
meters of Martin’s broth which had been handled as follows: 
Nine hundred fifty cubic centimeters of Martin’s broth neutral 
in reaction to litmus paper were inoculated with 50 cubic centi- 
meters of defibrinated virulent blood from carabao 3257 and 
kept in the incubators at 37° C. for twenty-four hours. There 
was considerable sediment in the broth at the time it was injected, 
but no microscopical examination was made to ascertain if it 
was free from visible microorganisms. 

On the evening of October 12, this animal ate very little and 
appeared uneasy. On the morning of October 13, it presented 
a slightly accelerated respiration, lay down a greater part of 
the time, and did not eat. During the afternoon of this day, 
the animal appeared very sick, and toward evening lay in a 
sprawled out position. It was observed that several quick res- 
pirations would be taken in succession, after which the animal 
would hold its breath for some time, as if in great pain. It 
could not get upon its feet, and was not able even to hold its head 
up for any length of time. 

On account of the critical condition, the animal was killed at 
5 p. m., which was twenty-six hours and a half after it had 
received the injection. Post-mortem examination revealed 
marked cedema at the points of inoculation, hemorrhagic in 
places. The duodenum was slightly congested; otherwise the 
internal organs presented no appreciable change. 

The temperature had remained stationary at 38°.8 C. through- 
out the experiment. 

Sections were made from parts of the cedematous tissue. The 
tissue spaces were filled with rod-shaped organisms, which in- 


_ dicated that the medium evidently had been contaminated with 


262 The Philippine Journal of Science 1914 


some organism other than that of rinderpest. This contamina- 
tion may have had a marked influence upon the symptoms which 
the animal presented, and it cannot be stated with any certainty 
that toxin derived from the rinderpest organisms, as stated by 
Baldrey, was the cause of the condition of the animal at the 
time it was killed. 

Experiment 4.—As soon as bull 3222 was killed, 50 cubic cen- 
timeters of the heart blood were injected subcutaneously into 
bull 3213. This animal suffered no ill effects from the injection, 
but was afterward proved to be susceptible to rinderpest. This 
shows that whatever agent caused the condition of 3222, it was 
not present in sufficient quantity in the heart blood to produce 
any ill effects upon another animal. 

EHxperiment 5.—The animal used was susceptible Batanes bull 
3286. On October 24, 1911, at 9 a. m., this animal received by 
subcutaneous injection, 700 cubic centimeters of culture broth 
which had been handled as follows: Nine hundred fifty cubic 
centimeters of Martin’s broth of neutral reaction to litmus paper 
were inoculated with 50 cubic centimeters of defibrinated virulent 
blood from bull 3223 and kept in the incubator at 37° C. for 
twenty-four hours. 

On October 27, the animal was noticed to be voiding a large 
quantity of blood in the urine. On the evening of October 28, 
it appeared to be very sick, refused food and water, and stood in 
a hunched up position with staring coat and with the head down. 
Respiration was rapid and catchy, the animal uttering a low 
grunt at the beginning of each expiration. There was a marked 
cedematous swelling along the abdomen and sides of the body. 
At 11 a. m., the animal lay in a sprawled out position with its 
head flat on the floor, presented marked symptoms of asphyxia, 
and frequently struggled. Death occurred at 12.30 p. m. 

Post-mortem examination revealed a slight congestion of the 
duodenum. Other parts of the intestinal tract presented no 
abnormal appearances. The kidneys were enlarged, markedly 
congested, contained numerous small hemorrhages, and were 
very friable in texture. The capsule was easily removed. The 
liver was congested and very friable. The subcutaneous tissue 
was markedly cdematous in the vicinity of the points of in- 
oculation and in the pendent portions of the body. 

This animal had a rise in temperature to 40° C. on the third 
and to 40°.6 C. on the fourth day after receiving the injection. 

Sections made from the cedematous tissue revealed numerous 


| 
} 
f 


mB, 3 Boynton: Cultivation of Rinderpest Virus 2963 


rod-shaped organisms in the tissue spaces similar to those found 
in the cedematous tissue of bull 3222. Hence, the symptoms and 
death may have been due to the products formed by these or- 
ganisms instead of the toxin formed by the rinderpest virus. 

Experiment 6.—This experiment was conducted similarly to 
the former ones with the exception that the broth was faintly 
alkaline in reaction to litmus paper. The animal used was sus- 
ceptible Batanes bull 3211. This animal was injected subcuta- 
neously at 2.30 p. m. on October 27, 1911, with 700 cubic 
centimeters of Martin’s broth which had been treated as follows: 
Nine hundred fifty cubic centimeters of Martin’s broth were 
inoculated with 50 cubic centimeters of defibrinated virulent 
blood from carabao 3258 and incubated at 37° C. for twenty-four 
hours. 

The animal showed a rise in temperature on the morning of the 
fifth day after injection, and the temperature remained above 
40° C. until the day of death. On the seventh day, the animal 
developed a diarrhcea, on the eighth day refused food, and death 
occurred late in the afternoon of the ninth day. Post-mortem 
examination revealed a marked congestion of the fourth stomach 
and duodenum. The cecum was congested and hemorrhagic in 
places. The gall bladder was distended and congested, perito- 
nitis was present, and there were erosions in the mouth. From 
the symptoms and post-mortem findings, one would be justified 
in concluding that this animal died from a typical attack of 
rinderpest. 

The results obtained from this experiment would lead one 
to infer that the toxin had not developed to such an extent 
as to have any vital effect upon the animal and that the virus 
of rinderpest had not been killed by the incubation nor its 
virulence depreciated in any respect. 

Experiment 7.—This experiment was a duplicate of the 
preceding one. The virulent blood used to inoculate the medium 
was taken from carabao 3245. 

The animal used in this experiment was susceptible Batanes 
cow 3294. This animal was injected subcutaneously at 2 p. m., 
October 31, 1911, with 900 cubic centimeters of the inoculated 
medium which had been incubated twenty-four hours at 37° C. 

No abnormal symptoms were present until the afternoon 
of the fifth day after injection, when the animal showed a 
pronounced rise in temperature. The temperature remained 
abnormally high until the day of death. On the eighth day, the 
animal developed a diarrhcea and ate but little. On the ninth 

1268845 


264 The Philippine Journal of Science 1914 


day, it presented a profuse diarrhcea and refused food. On the 
tenth day, bloody mucous casts were passed and the animal 
lay down most of the time, dying in the evening. 

Post-mortem examination revealed erosion ulcers in the mouth; 
marked ulceration and congestion of the fourth stomach and 
duodenum; and congestion, ulceration, desquamation, and 
hemorrhagic areas in the cecum, colon, and rectum. Blood- 
stained mucous casts were present in the lower colon and rectum. 
Peritonitis and distention of the gall bladder were noted, and 
the vagina was markedly congested. 

The incubation period, symptoms, and post-mortem findings 
lead to the conclusion that this animal died of a typical attack 
of rinderpest. Evidently the virus was not destroyed nor its 
virulence attenuated to any appreciable extent by the medium 
or incubation for twenty-four hours. 

The results obtained from this experiment indicate that the 
toxin, if produced in the culture medium, was not present in 
sufficient quantity to have any effect upon either the virus or 
the animal injected. 

Experiment 8.—This experiment was carried on simultaneously 
with the preceding one, and was similar with the exception 
that carabao 3262, supposedly susceptible, was used instead of 
a bull. 

The animal was injected subcutaneously at 2.30 p. m., October 
31, 1911, with 900 cubic centimeters of Martin’s broth, which 
had been inoculated with 50 cubic centimeters of defibrinated 
virulent blood from carabao 3245 and incubated for twenty-four 
hours at 37° C. 

The injection of the incubated broth had no immediate effect 
upon the animal. It did not contract rinderpest, whereas the 
animal in experiment 7, receiving medium inoculated with blood 
from the same animal, contracted the disease. This carabao 
was later injected with 50 cubic centimeters of virulent blood, 
but did not contract rinderpest. This leads to the conclusion 
that this animal was immune to the disease at the time of the 
experiment. 

From these results, it will be noted that the culture had no 
visible effect upon this animal. 

Experiment 9.—This experiment varied slightly from the 
preceding one in that the inoculated medium was incubated 
forty-eight hours, which gave double the time for the elaboration 
of toxin. 

The animal used in the experiment was susceptible Timor 


en 


1X, B, 3 Boynton: Cultivation of Rinderpest Virus 965 


bull 3293. It was injected subcutaneously at 3 p. m., November 
10, 1911, with 900 cubic centimeters of Martin’s broth neutral 
in reaction to litmus. The medium had been inoculated with 
50 cubic centimeters of defibrinated virulent blood from carabao 
3241, and was incubated forty-eight hours at an average tem- 
perature of 36° C. 

On the morning of the fourth day after injection, the first 
rise of temperature occurred, which was followed by a tempera- 
ture of 40° C. in the afternoon. The temperature remained 
febrile until the day of death. Diarrhcea developed on the tenth 
_ day, at which time the animal displayed inappetence. On the 
eleventh day, the diarrhoea was profuse and continued until death, 
which occurred on the morning of the thirteenth day after 
injection. 

Post-mortem examination of this animal revealed erosion 
ulcers in the mouth; ulceration and congestion of the fourth 
stomach; marked congestion and hemorrhages in the duodenum, 
cecum, colon, and rectum; and peritonitis and emphysema of 
the lungs. 

In symptoms and lesions, this animal presented all the ap- 
pearances of having died of rinderpest. Evidently, if any toxin 
was formed after the forty-eight-hour incubation period in 
the culture medium, it was so weak that it had no appreciable 
effect upon the animal injected and did not kill the virus in the 
medium or weaken its virulence. 

Experiment 10.—This experiment differed from the preceding 
one in that the medium used was slightly acid in reaction to 
litmus and susceptible Timor cow 3292 was employed. The cow 
was injected subcutaneously at 3.15 p. m., November 10, 1911, 
with 900 cubic centimeters of Martin’s broth, which had been 
inoculated with 50 cubic centimeters of defibrinated virulent 
blood from carabao 3241 and incubated at approximately 36° C. 
for forty-eight hours. 

The animal showed no immediate ill effects from the injection, 
and did not show a rise in temperature nor display any of 
the symptoms of rinderpest up to the thirteenth day after 
injection. 

The susceptibility of this animal was not proved, since it was 
accidentally killed before being tested. 

From the results of this experiment, it is evident that if there 
was any toxin formed in the medium it was so slight as to 
have no ill effect upon the animal. It also appears as if the 
acid medium killed the virus, although there is no experimental 


266 _ The Philippine Journal of Science 1914 


proof that this animal was susceptible. The fact of its suscep- 
tibility may be taken for granted, since it was one of fifteen 
Timor animals which were used at the laboratory and the other 
fourteen contracted rinderpest and died. ' The virulence of the 
blood used for inoculating the medium was checked up as shown 
in the preceding experiment. 

Experiment 11.—In this experiment, 950 cubic continietend 
of a 5 per cent potassium citrate solution were used instead of the 
Martin’s broth. This solution was inoculated with 50 cubic 
centimeters of defibrinated virulent blood from carabao 3241 
and incubated at approximately 36° C. for forty-eight hours. 

The animal used was susceptible Timor cow 3296. It was 
injected subcutaneously at 3.30 p. m., November 10, 1911, with 
900 cubic centimeters of this culture. 

The animal suffered no immediate ill effects from this injec- 
tion, and did not contract rinderpest. It was inoculated later 
with virulent blood, and was proved susceptible to rinderpest. 

It is concluded from these results that either no toxin was 
formed in the potassium citrate solution or it was formed in such 
a small quantity that it produced no ill effects upon the animal. 
Also, the virus was not able to survive in it forty-eight hours 
under the conditions existing. The virulence of the blood used 
in this experiment for inoculating the medium was checked up 
as the results show in experiment 9. 

Experiment 12.—In this experiment, Martin’s broth was 
strongly alkaline in reaction to litmus paper. The animal used 
was susceptible Timor cow 3297. It was injected subcutaneously 
at 3.45 p. m., November 10, 1911, with 900 cubic centimeters 
of Martin’s broth, which had been inoculated with 50 cubic 
centimeters of defibrinated blood from carabao 3241 and 
incubated at approximately 36° C. for forty-eight hours. 

This animal presented no immediate ill effects from the 
injection. It developed a temperature on the sixth day after 
injection, diarrhoea on the tenth day, inappetence on the twelfth 
day, and died during the following night. 

Post-mortem examinations revealed erosion ulcers in the mouth 
and a slight congestion of the fourth stomach and duodenum. 
The cecum and colon were but slightly changed. The rectum 
was slightly congested and hemorrhagic, and a slight peritonitis 
was present. The lesions were not so pronounced as those noted 
in the preceding autopsies. 

From the results obtained from this experiment it is evident 


1X, B, 3 Boynton: Cultivation of Rinderpest Virus 267 


that if there was any toxin formed in the medium after a forty- 
eight-hour incubation period it was so small in amount as to 
have no immediate ill effect upon the animal in question. From 
the incubation period, together with the symptoms and lesions 
presented upon autopsy, it is evident that the virus was not 
killed and that it had lost practically none of its virulence. 

In addition to the experiments recorded in this article, three 
animals were injected in a similar manner with virulent rinder- 
pest blood which had been incubated for three days. In each 
case, no ill effect followed the injection. It was concluded that 
in none of these cases was a harmful amount of rinderpest 
toxin produced, nor was there evidence that the rinderpest virus 
survived in the medium. 


CONCLUSIONS 


1. In the case of the two animals in experiments 3 and 5, 
that died in less time than the incubation period of rinderpest, 
after injection of the Martin’s broth culture, the autopsy find- 
ings of the tissues indicated death from a bacterial infection 
and not from rinderpest. All evidence points to the conclusion 
that the Martin’s broth employed in these two cases was con- 
taminated by bacteria prior to injection in the animals. The 
results are attributed to poor aseptic technique, and greater 
care in the subsequent inoculations, where no such toxzemias 
were induced in the injected animals, support the conclusion. 

The symptoms, lesions, and other circumstances stated by 
Baldrey resemble the results obtained in the two animals in 
question, and there is justification for belief that his results 
were due to the same cause. 

2. In all the other animals injected with mixtures of blood 
and culture medium after incubation, no immediate ill effect 
followed, in either susceptible or immune animals. 

3. With the exception of the animals noted in experiments 
3 and 5, all those injected with the so-called twenty-four- and 
forty-eight-hour cultures of rinderpest in neutral or alkaline 
Martin’s broth contracted rinderpest after the usual incubation 
period and died. These observations do not support Baldrey’s 
belief that there occurs a rapid formation of rinderpest toxin 
in the broth during the twenty-four hours with resulting death 
of the virus. The experiments have included tests of Martin’s 
broth after incubation as long as seventy-two hours. 

Rinderpest virus does die in Martin’s broth culture after 


268 The Philippine Journal of Science 19u4 


incubation for seventy-two hours, but there is no evidence that 
rinderpest toxin was formed, much less that rinderpest toxin 
caused the death of the virus. 

5. The experiments reveal the fact that rinderpest virus will 
survive in neutral or alkaline Martin’s broth at 37° C. for at 
least forty-eight hours, but not for seventy-two hours. Two 
cases were tested at twenty-four hours, 2 at forty-eight hours, 
and 3 at seventy-two hours. 

6. Rinderpest virus kept in acid Martin’s broth or in 5 per 
cent potassium citrate solution did not survive after forty-eight 
hours at 37° C. 


KIDNEY-WORM INFESTATION OF SWINE IN THE PHILIPPINE 
ISLANDS WITH SPECIAL REFERENCE TO THE 
; PATHOLOGICAL CHANGES 


By WILLIAM HUTCHINS BOYNTON 
(From the Veterinary Division, Bureau of Agriculture, Manila, P. I.) 


Three plates 
INTRODUCTION 


Kidney-worm disease of swine is caused by an infestation with 
Stephanurus dentatus Diesing, 1839.1. The infestation is char- 
acterized by muscular pains, tenderness to pressure over the 
kidneys, weakness, loss of appetite, emaciation, and partial or 
complete paralysis of the hind quarters. The parasites may be 
located in the fat surrounding the kidneys, in the ureters, and en- 
cysted in the kidneys, liver, spleen, lymph glands, and muscles and 
connective tissues in the region of the kidneys; also, they may 
be found free, slightly embedded, or encysted in the connective 
tissue of both peritoneal and thoracic cavities. 

From the reports at hand, kidney-worm infestation appears 
to be more prevalent in subtropical and tropical climates than 
in the colder latitudes. It has been found in the United States, 
especially in the southern part. It is also present in South 
America, Australia, Asia, and in the Philippine Islands. 

Observations upon the pathological changes induced by the 
worm in pigs imported into the Philippines form the basis of this 
paper. 

The following description of the worm is taken from a bulletin 
written by Tayler? on this subject. 

Sclerostoma.—Body cylindrical, tapering but slightly at each end; mottled 
in color; mouth terminal, circular, provided with six papille, of which the 
dorsal and the ventral are more prominent than the four submedian, and 
with numerous cilialike structures, about 35-40 in number. Cuticle thicker 
at extremities than in middle of body; transverse rings broader at extrem- 
ities (9 “) than in the middle (3 “) of body. Opening of cephalic gland 
on ventro-median line near anterior end of esophagus. Alimentary tract 


*Synonyms: Sclerostoma pinguicola Verrill, 1870. Lard worm of pigs. 
*16th Annual Rep. Bur. Animal Ind. for 1899. Washington (1900), 612- 
637. 
269 


270 The Philippine Journal of Science 1914 


with distinct regions; buccal cavity ball-shaped, with ten enlargements at 
base; esophagus distinct, Indian-club shaped; stomach intestine greatly 
convoluted. 

Male: 25 to 37 mm. in length by 14 mm. in width. Anus at caudal 
extremity. Inconspicuous rectangular bursa; lobes 6; dorsal 2, latero-dorsal 
2, latero-ventral 2. Hach half of bursa, 9 rays, arranged: dorsally 3, 1, 
dorso-lateral 3, ventro-lateral 2. Spicules 2, equal, saberform, ca. 0.8 mm. 
long; vesicule seminales 2; vasa deferentia 2, and testicles 2; each vas 
deferens with one testicle forming a long, white, convoluted tube about 
26 mm. in length. 

Female: 37 to 40 mm. in length by 2 mm. in breadth. Tail curved, 
obtuse, except for a conical tip. Lateral cuticular wings at caudal extrem- 
ity. Anus ventral, 0.64 mm. from caudal extremity. Vulva 1 mm. forward 
from anus. Uteri 2, vagina bicornate. Ovaries 2, oviducts 2, each ovary 
with one oviduct forming a long, white, convoluted tube ca. 280 mm. in 
length. Oviparous; eggs oval, 56 » by 100 4; shell thin and smooth. 


THE INFECTION IN THE PHILIPPINES 


I am indebted to Dr. R. W. Newcomb, veterinarian in charge 
of meat inspection in the Manila matadero, for data showing 
the extent of infestation and distribution of the disease among 
swine slaughtered in Manila. About 2,000 hogs were examined, 
of which very nearly 50 per cent were found infested. Of the 
animals slaughtered, approximately 20 per cent came from 
Batangas and Bulacan Provinces, respectively, 15 per cent were 
raised in Manila, while 10 per cent came from Cavite and Rizal 
Provinces, respectively. The remainder came from Zambales, 
Tarlac, Pampanga, and Nueva Ecija. 

Dr. A. S. Shealy examined 43 animals in the same matadero, 
and found 50 per cent to be infected. A collection of lesions 
containing these worms was furnished to me, and has been 
useful in the preparation of this paper. 

I have been afforded the opportunity of observing the effects 
of kidney-worm infestation among imported swine. The group 
of 20 pigs forming the nucleus of the drove consisted of 5 from 
Texas, 138 from Australia, and 2 from New Zealand. The im- 
portation began in 1906 with 5 from Texas. 

The first definite diagnosis of kidney-worm infestation was 
found in a Berkshire sow imported from Australia on December 
38,1909. This animal died on December 8, 1911, two years after 
its arrival at the farm. A Berkshire boar (No. 10) imported 
from Australia in 1907 died on March 30, 1910. Upon inquiry, 
I was informed that this animal became paralyzed in its hind 
quarters shortly before death, which is a symptom of kidney- 
worm infestation. 

From December 3, 1911, to April 22, 1912, covering a period 


ee ae — ————_ —— --— 


— 


IX, B, 3 Boynton: Kidney-worm Infestation of Swine Val 


of four months and nineteen days, 6 Berkshire sows died, and 
all showed marked lesions of kidney-worm infestation, which 
was undoubtedly the cause of their death. 

The animals were placed under the best sanitary conditions, 
being housed and kept on cement floors which were washed 
daily. The refuse was carried away through a sewer. Each 
compartment had a small runway for the animals to get out 
on the ground. They were not crowded, for very seldom over 
5 or 6 animals were kept in the same pen at one time. None 
but the boar came in contact with native pigs. Shortly before 
a sow was ready to give birth to young, she was placed in a pen 
alone and kept there until the litter was old enough to wean. 
The only time during which a large number of pigs were together 
was after they were weaned. Those of nearly the same age 
were placed in a large pen by themselves. When they reached 
about 5 months of age, the animals to be kept for breeding pur- 
poses were taken out and the rest were disposed of. 


SYMPTOMS 


The first noticeable symptom is a stiffness of the movements 
of the animal as if it were suffering from muscular pains. This 
is especially perceptible in the hind limbs. As the disease pro- 
gresses, the animal becomes lame and weak in the loins. During 
this stage, it lies down most of the time and does not rise to its 
feet unless urged. In some cases the animal loses partial or 
complete control of the hind quarters a few days before death. 
There is tenderness over the kidneys, as wincing occurs when 
pressure is brought upon these regions. The animal usually 
eats fairly well up to within a few days of death, when it refuses 
food entirely. Emaciation is generally observed, but the reverse 
was noticed in 2 animals which were in good condition at the 
time of death. Ascites was present in 3 of the 6 cases examined. 


LOCATION OF THE WORMS 


In the 6 animals examined, worms were observed in the follow- 
ing locatiens: Both free upon and deep in the fat surrounding 
the kidney, in the ureters, in the pelvis of the kidney (Plate 
I, fig. 4, ¢), and deep in the cortex and medulla of the kidney 
(Plate I, fig. 4, d); free in the peritoneal cavity, and often 
penetrating the peritoneum; free on the surface of the liver, 
embedded in the connective tissue around the large blood vessels 
of that organ, and in the liver tissue, either in accumulations or 
slightly embedded under the capsule, and deep in both portal 


22 The Philippine Journal of Science 1914 


and mesenteric lymph glands; and in great numbers in the con- 
nective tissue along the back, especially in the neighborhood of 
the kidneys. In 2 cases, the connective tissue along the spinal 
column extending from the coccyx to the diaphragm was a mass 
of worm tracts, most of which contained one or two worms and 
purulent material. In 3 cases, worms were found free in the 
thoracic cavity and embedded in the pleura. One worm was pene- 
trating the lung tissue. Some were embedded in the diaphragm. 
Thus it will be seen that, when an animal dies of kidney-worm 
infestation, the disease presents a generalized instead of a 
localized aspect. 

Several writers state that the worm is never found in the in- 
ternal structure of the kidney. This may be true in those cases 
noticed in the abattoirs where the infestation is not of long 
standing, but if one examines animals which have died of the 
disease he will find the kidney structure to be invaded in a 
majority of cases. 


MORBID ANATOMY 


One of the most prominent changes to be noted at autopsy is 
the rapidity with which the internal organs undergo post-mortem 
changes. Four of the 6 animals were examined shortly after 
death. The viscera of these animals had a disagreeable odor as 
if decomposition was well advanced. Changes in the various 
organs were observed as described below. 


LIVER 


In every case, the liver was enlarged, soft, and of a dark 
purplish color. The borders of the lobes instead of tapering 
down to a rather thin edge were distended and rounded. In 
some instances, collections of gas bubbles were noticed under 


the capsule, suggesting that putrefaction was taking place 


rapidly. The dependent portions of the lobes of the liver were 
of a uniform dark purplish color, and the individual lobules were 
not distinguishable through the capsule. The upper portion of 
the lobes had a mottled appearance. The majority of the lobules 
in some cases were dark purple, while the remaining lobules varied 
from a purplish tint to gray-brown. Upon section, the paren- 
chymatous tissue was soft and bulged out on the incised surface, 
and the borders of the lobules were indistinguishable. The lobes 
adhered to each other by a gray, feltlike fibrinous exudate. 
This was especially prominent in the dependent portions. In 3 
cases, the liver was adherent to the diaphragm, sternum, and 
stomach. 


IX, B, 3 Boynton: Kidney-worm Infestation of Swine 273 


Several parietal thrombi were found in the branches of the 
hepatic veins (Plate I, fig. 2, 6). The largest one noticed 
measured 1.9 centimeters in length, and filled approximately half 
of the vessel. These thrombi are undoubtedly the principal 
agents which cause the enormous passive congestion in the liver 
by damming back the blood in the hepatic system. 

The walls of the main branch of the portal vein and vena cava 
in some cases were considerably thickened, and possessed worm 
tracts which were surrounded by fibrous connective tissue and 
which contained worms or a mass of purulent material. These 
worm tracts furthermore extended into the liver substance, re- 
placing the parenchymatous with fibrous tissue. In 2 cases, large 
accumulations of worm tracts were found in the regions where 
the lobes join and on the posterior surface. Each of these 
accumulations was surrounded by considerable connective tissue. 
The largest of them measured 10.5 by 4.5 centimeters, and hence 
extended a considerable distance into the liver substance. In the 
immediate vicinity of these accumulations, isolated tracts were 
found. The individual tracts varied from 1 to 2 millimeters in 
diameter, and the walls were made of a gristlelike substance 
which in turn was surrounded by connective tissue. 


KIDNEYS 


The kidneys like the liver showed a marked deviation from 
the normal. A photograph of a kidney from pig A-18 is shown 
in Plate I, fig. 3. This kidney measured 15.5 centimeters in 
length and 6 centimeters in diameter. A retention cyst (Plate I, 
fig. 3, a) was present on the external surface of the anterior end, 
and measured 3 by 2 centimeters. It was somewhat triangular in 
shape, the apex projecting toward the pelvis of the kidney. The 
outer wall of the cyst was denuded of parenchymatous tissue, 
was composed of tough, fibrous connective tissue, and was set 
rather deep into the substance of the cortex. It is questionable 
whether the worms were the cause of the cyst formation. How- 
ever, retention cysts were present in the kidneys of 3 of the 
6 hogs examined. Scattered over the surface of the kidney were 
several uneven dark-purplish indentations, which were appar- 
ently the scars of small healed abscesses. The surface of the 
kidney was not uniform in color, but presented the appearance 
of being injected with fine, irregular, reddish brown lines forming 
a multitude of grayish brown blotches, averaging 1 millimeter 
in diameter. The capsule of this kidney stripped easily. The 
retention cyst contained a turbid semiviscid fluid. The internal 
surface of the wall of the cyst was composed of smooth, fibrous 


274 The Philippine Journal of Science — 1914 


connective tissue. On section, the cortical substance was gray, 
with brownish red streaks and blotches following the uriniferous 
tubules down into the medulla. The medullary substance was 
similar in appearance to the cortex. The papille had a con- 
gested appearance, and were slightly swollen. The pelvis con- 
tained a large amount of gelatinous and purulent material. Two 
worms were found embedded in this purulent material. 

The left kidney from pig A-21 measured 11.5 by 5.5 centi- 
meters. A retention cyst was present on the external surface of 
the posterior end, and measured 1.7 by 1.8 centimeters. It was 
deep set in the tissue, and was covered by a thin fibrous capsule. 
This kidney had several deep notches on its surface, apparently 
scars from former abscesses or cysts, which gave the kidney a 
shrunken, uneven appearance. It was of a uniform grayish 
brown color, and the capsule was easily removed. 

The right kidney from pig A-14 measured 12 by 5.5 centi- 
meters. The capsule was easily removed. The kidney was of 
a bluish gray color, and was of soft consistence. The external 
surface was perforated with numerous small holes, averaging 
about 0.5 millimeter in diameter, which gave the kidney exter- 
nally a porous appearance. This was due undoubtedly to rapid 
post-mortem changes. The inner surface was somewhat flat- 
tened, having a large scar near the center. Two worms pro- 
truded from the cut end of the ureters. A large amount of 
yellowish white purulent material escaped from the ureter when 
it was cut. 

On sectioning the kidney through its longitudinal axis, the 
internal structure, especially the cortex and medulla, had the 
appearance of a sponge. It was perforated with holes and pits, 
varying from 0.5 to 3 millimeters in diameter. (Plate I, fig. 4, a 
and b.) Practically all the parenchymatous tissue was destroyed, 
leaving merely the connective tissue framework. This appear- 
ance was particularly prominent in the posterior two-thirds, while 
the anterior third was badly affected, but not to such a marked ex- 
tent. The cortex and medulla were gray, except on the borders 
of the papillz, where the medulla was brownish. Many of these 
changes were undoubtedly due to rapid decomposition. The 
papilla were brownish in color, and contained a few pits or 
vacuoles. ; 

The calix minor and calix major were brown and considerably 
distended. Three worms were coiled up in the pelvis of the kid- 
ney, and, as mentioned above, there were 2 in the ureters, thus 
bringing to view 5 worms in this particular kidney in one section. 


1g B, 8 Boynton: Kidney-worm Infestation of Swine 9795 


A section through the apparent scar on the flattened inner 
surface of this kidney revealed an area 2.8 by 1.9 centimeters, 
composed of fibrous connective tissue, and containing 9 worm 
tracts varying from 3 to 5 millimeters in diameter. Five worms 
were found in these tracts. This area was located principally in 
the medulla, a part of it extending up into the cortex. In all, 8 
worms were found in this kidney and 2 in its ureter. 


LYMPH GLANDS 


In every case examined, the portal and mesenteric lymph 
glands were enlarged, one of average size measuring 5.5 by 
2.3 centimeters. As a rule, in the early stages they were soft 
in consistence, but became indurated in the older lesions. The 
external surface of the glands which had been infested for some 
time and had become indurated was very uneven. Small pro- 
tuberances extended up from the surface of the glands, where 
the worms had embedded themselves in the capsule. 

On sectioning, the indurated glands were cut with some dif- 
ficulty. The cut surface presented different appearances, de- 
pending upon the extent to which changes had taken place. 

In the early stages, the glandular structure had a congested 
reddish appearance throughout. However, this was most prom- 
inent along the borders of the capsule and the trabecule. In 
the next stage, the capsule and trabecule were considerably 
thickened. The internal glandular structure was colored gray, 
except along the border of the thickened capsule and trabecule, 
where it was dark red from the congestion and blood pigment 
present. 

Tracts either containing worms or filled with purulent mate- 
rial were found especially in the thickened capsule and sometimes 
extending into the larger trabecule. 

A few glands were found in which practically the entire paren- 
chymatous structure had become transformed into connective 
tissue intermingled with worm tracts which contained either 
worms or a mass of round cells and tissue detritus. 


ABDOMINAL CAVITY 


Ascites was present in 3 of the cases examined; namely, A-21, 
A-27, and A-18. In A~-21 there were between 3 and 4 liters of 
blood-stained fluid. The small and large intestines were one 
mass of adhesions. The intestinal walls were cedematous, and 
the mucous membrane was highly congested throughout. 

In the other 2 cases there was not as much ascitic fluid 


276 The Philippine Journal of Science 1914 


present; however, the intestines were cedematous and adherent 


as in A-21. 
LUNGS 


More or less pleurisy was present in all 6 cases. The lobes 
adhered to each other and in some instances to the thoracic wall. 
In 8 cases, kidney worms were found in the thoracic cavity, but, 
as the bronchioles in every case contained large numbers of 
worms belonging to the genus Metastrongylus, it is rather diffi- 
cult to decide whether the pleurisy was caused by them or by 
Stephanurus dentatus. 


MICROSCOPICAL EXAMINATION OF INFESTED ORGANS 


The tissue under consideration was fixed in both Zenker’s fluid 
and formalin, and paraffin as well as frozen sections were made. 
The sections were stained with Ehrlich’s acid hematoxylin and by 
Van Gieson’s method, using Ehrlich’s acid hematoxylin and 
picrofuchsin. Sections revealed the following conditions: 


KIDNEYS 


The kidney tissue showed a pronounced passive congestion. 
The veins and capillaries between the tubules in both cortex and 
medulla were distended with blood. In some instances, a con- 
siderable amount of congestion was present in the glomeruli. 
Since the arteries were practically empty, undoubtedly the con- 
gestion of the glomeruli was due to the enormous passive conges- 
tion which dammed the blood back in them. 

Parenchymatous degeneration was present in all stages of 
development and scattered diffusely throughout the cortical and 
medullary substance. Numerous casts were present in the con- 
voluting and collecting tubules (Plate II, fig. 2, a). These casts 
had all the appearances and staining reactions of colloid material, 
for instance giving a homogeneous orange-red color, when 
stained with hematoxylin and picrofuchsin. 

Numerous parenchymatous cells were noticed undergoing 
colloid degeneration. They presented different appearances, 
depending upon the extent to which the degeneration had taken 
place. Inthe earliest stage noticeable, the cell body took a lighter 
stain and was very finely granular. In the next stage, small 
spherical droplets, which took an orange stain with hematoxylin 
and picrofuchsin (Plate II, fig. 3, a), could be noticed scattered 
throughout the cell body. As the degeneration progressed, the 
droplets became larger and finally occupied the entire cell body 
(Plate II, fig. 3, c). Finally, they burst out into the lumen of 
the tubule, and eventually coalesced to form casts (Plate II, fig. 


1X, B, 3 Boynton: Kidney-worm Infestation of Swine QT eT: 


2, b). The nuclei retained their staining power remarkably, as 
will be noticed in Plate II, fig. 3, d. Although the cell body had 
become completely broken up into droplets, the nucleus stained 
almost as perfectly as in a healthy cell. The nuclei did not 
become destroyed until after they had flowed into the lumen of 
the tubule. 

All the parenchymatous tissue was not affected in this manner. 
The degeneration occurred in areas, involving from 4 to 8 or 
more tubules, and was more prominent in the cortex than in the 
medulla. Frequently, there were accumulations of round cells 
which had the appearance of lymphoid tissue and tock the 
hematoxylin stain deeply. These cells were either scattered 
through, or were in the immediate vicinity of, the degenerating 
areas. 

An occasional glomerulus was found, in which the space be- 
tween the outer and inner capsule was filled with homogeneous 
colloid-appearing material. As the glomerular tissue and cap- 
sule showed no apparent degeneration, this colloid material must 
have been forced into the cavity through the constricted neck 
of the convoluted tubules. The marked passive congestion was 
undoubtedly instrumental in the retrogade flow of this material. 

Sections made from the kidney represented in Plate I, fig. 4, 
showed that there was practically a complete destruction of the 
parenchymatous tissue in both the cortex and medulla. The 
lining connective tissue framework of the tubules could be seen 
practically denuded of parenchymatous tissue. The spaces were 
either empty or filled with cell detritus mixed with a faintly 
staining, colloidlike substance, which showed complete destruc- 
tion of the functional tissue in that particular specimen. 


LIVER 


The most striking appearance in the liver on microscopical 
examination was the marked passive congestion present. This 
varied in degree in different portions of the organ. 

As a rule, sections made from the upper portions of the lobe 
showed the portal vein to be distended with blood, practically 
all the interlobular capillaries were congested, and the central 
vein was distended. The liver cells generally showed slight 
parenchymatous degeneration. In the interstitial tissue, there 
was usually an abnormal number of leucocytes present, especially 
small lymphocytes, and to a lesser extent eosinophiles. Leuco- 
cytes were also seen in the interlobular capillaries, working 
their way through the capillary walls and invading the individual 
liver cells. ’ 


278 The Philippine Journal of Science 1914 


The next and most spectacular stage of congestion was noted 
in the lower portions of the lobes. The portal veins were 
usually distended with blood, the interlobular capillaries around 
the edges of the lobules were markedly congested, while in 
the central portion of the lobules around the hepatic vein (Plate 
II, fig. 1, a) the congestion reached such a degree as to cause 
rupture of the capillaries. These markedly congested and 
hemorrhagic portions varied from one-fourth to three-fourths 
of the area of the involved lobules. The parenchymatous tissue 
in these regions had practically disappeared. Now and then 
a liver cell was present which had not as yet undergone necrosis. 
The parenchymatous tissue around these areas was found in 
all degrees of degeneration, being mildest near the capsule of 
the lobules. An abnormal number of round cells and eosino- 
philes were found in the capsular tissue, also penetrating into 
the parenchymatous tissue, and surrounding the individual liver 
cells. In a few instances, bile ducts were noticed undergoing 
proliferation in this area. 

The next stage noticed was that in which the lobules had 
become necrotic and were filled with blood from hemorrhages. 
These areas were most frequently met with at the extreme 
dependent portions of the lobes (Plate I, fig. 1, a). The 
hemorrhagic areas contained large numbers of round cells and 
eosinophiles. The capsule was congested and infiltrated with 
leucocytes. The connective tissue of the capsule showed signs 
of degeneration, especially infiltration with a granular, fibrinous 
exudate. 

In those areas of the liver which the worms had invaded, 
marked pathological changes were encountered. Sections of 
worms were frequently found. Surrounding the worm was an 
accumulation of leucocytes and cell detritus, external to which 
was a fibrous connective tissue wall. More frequently the track 
of the worm was found, which was represented by a round or 
oblong mass of cell detritus mixed with round cells and fibrin, 
and surrounding this mass of material was the fibrous connective 
tissue wall. 

In the immediate vicinity of these invaded areas, there was 
marked development of connective tissue. In many instances, 
the interlobular connective tissue had developed to such an extent 
as to crowd the lobules out of existence. In some instances, 
lobules were found varying from three-fourths to one-fifth of their 
normal size. The liver cells were atrophied, from the pressure 
brought on them by new-forming connective tissue. The most 


a 


IX, B, 3 Boynton: Kidney-worm Infestation of Swine 279 


embryonic type of connective tissue and the youngest blood 
vessels were found around the edges of the lobules, while the 
connective tissue which extended out from the edges appeared 
to be older; this showed that it was encroaching upon the 
parenchymatous tissue and crowding it out. Large numbers 
of round cells and eosinophiles were present along the borders 
of the lobules, and a few were scattered throughout the new- 
forming connective tissue. 

From the foregoing, it is evident that the worm produces 
an irritation which gives rise to proliferation of connective at 
the expense of parenchymatous tissue, the former causing 
pressure upon, and atrophy and degeneration of, the latter. 

There was also marked proliferation of the bile ducts in these 
areas. They developed in some instances to such an extent 
as to present the appearance of an adenoma. In those areas 
in which the parenchymatous tissue had been completely obliter- 
ated by the intruding connective tissue, many of the numerous 
blood vessels which were formed during the development of 
the connective tissue had undergone thrombosis. Thrombi in 
all stages of organization were found. In many instances, 
these thrombi appeared as cords of connective tissue completely 
occluding the small vessels. 

In those areas which the worms had but recently invaded, 
the fibrous connective tissue capsule surrounding them was found 
in different degrees of development. The earliest stages showed 
merely a few cells of new-forming tissue, rich in blood vessels 
and infiltrated with a large number of leucocytes. In the older 
lesions, there was a well-defined, thick wall of connective tissue. 
The liver tissue around the newly infested parts showed a 
marked passive congestion. Also, the liver cells in these 
areas were shrunken to a perceptible extent from pressure 
caused by dilated capillaries, and they showed various grades 
of degeneration. 

LYMPH GLANDS 


Various appearances were noticed in the lymph glands accord- 
ing to the extent to which they had undergone pathological 
changes. The earliest change was an active congestion. The 
cortical follicles, especially in the neighborhood of the lymph 
sinuses, were markedly congested, and merged in many instances 
into hemorrhagic areas, The cortical substance was distended. 
The lymphoid tissue was scattered, and the intervening spaces 
were filled with blood. In the medullary portion of the glands, 
the tissue was distended and loose, except the lymphatic cords 

126334——6 


Veo e | eee 


280 The Philippine Journal of Science 1914 


which in most cases held their closely packed appearance. The 
medulla was likewise markedly congested, and in some instances 
hemorrhagic. 

The next stage showed a thickening of the capsule and 
trabecule, with pronounced congestion, hemorrhages, and even 
necrosis of the follicular substance. Thrombi were frequently 
found in the blood vessels. In Plate II, fig. 4, c, is shown a 
large thrombus in an artery located in the trabecule. This 
particular thrombus had almost occluded the vessel, and was of 
the mixed type. The vessel walls, b, and the trabecule, d, were 
thickened. 

In some of the glands thus affected, one or more worms were 
embedded in the capsule, passing down into the connective tissue 
of the trabecule. 

As the infestation persists, the worms work their way into 
the substance of the gland. They cause an irritation which 
brings on an active inflammation, resulting in the formation of 
a connective tissue capsule around the worm tract. In the 
course of time, the entire glandular substance is replaced by 
connective tissue, and the refuse is left by the worm. A section 
from the lymph gland is represented in Plate III, fig. 1, in 
which practically all the glandular substance has been replaced 
by worm tracts and fibrous connective tissue. 

Worms were found repeatedly in the glands. They were 
surrounded by leucocytes, most of which were small lymphocytes 
intermixed with cell detritus and fibrin. The tract wall was 
made up of proliferating connective tissue which contained 
numerous new-forming blood vessels, both single and ramified. 
Large numbers of round cells and eosinophiles were scattered 
throughout the proliferating connective tissue, and were especially 
abundant in the borders of the tissue next to the worm. 

A tract apparently occupied by a worm is shown in Plate 
III, fig. 1, a. It is composed of fibrin, degenerated cells, and 
small round cells. This tract is surrounded by a connective 
tissue capsule, b, which contains a large number of new-forming 
and ramified blood vessels, c. These vessels are practically all 
distended with blood. A large number of leucocytes and 
eosinophiles are scattered throughout the connective tissue. A 
worm tract which is of longer standing than the one shown in 
Plate III, fig. 1, a, is represented in Plate III, fig. 1, d. The 
contents of this tract are made up of fibrin, cell detritus, and 
leucocytes. There is a layer immediately around this (fig. 1, e) 
which takes a pinkish red stain with eosin, which suggests 


1X, B, 3 Boynton: Kidney-worm Infestation of Swine 281 


early calcification because of its homogeneous appearance and 
which contains a.large amount of cell detritus. Immediately 
around this zone is the capsule, f, made up of rather old and 
well-formed connective tissue. Hence it appears that this worm 
tract has been present for some time. A still older tract is 
shown in Plate III, fig. 1, g, in which the central part has become 
a granular mass of broken down and degenerated cells. The 
zone which bounds this central mass, h, stains a rather deep 
red with eosin; it is homogeneous, and has all the appearances 
of a calcified mass. The zone, 2, surrounding this is made up of 
old fibrous connective tissue which forms the wall of the tract. 


SUPPLEMENTARY OBSERVATIONS 


Since the completion of the preceding general description 
of lesions, two animals, Berkshire sows A-17 and A—25, have died 
of kidney-worm infestation. 

Animal A-17 was 4 years 8 months and 6 days old when she 
died. She aborted about three weeks prior to death, and had 
seemed unthrifty for some time. Death occurred in the evening 
of October 27, but the autopsy was not made until the following 
morning. 

Post-mortem changes were well advanced. The viscera had 
a pungent repulsive odor. The peritoneal cavity contained a 
large amount of blood-stained fiuid. The liver was markedly 
distended, soft, of a purplish color, and contained collections 
of gas bubbles under the capsule. The lobes were slightly 
adherent to each other at their dependent portions by a grayish 
feltlike exudate. 

The small intestines and especially the colon were cedematous 
and slightly adherent by a fibrinous exudate. The mesenteric 
lymph glands were enlarged and in some instances slightly 
indurated. 

The right kidney measured 16 centimeters in length, 9 centi- 
meters in width at its anterior end, and 6.5 centimeters in 
width at its posterior end. It was 5.5 centimeters in thick- 
ness at the anterior end and 4 centimeters at the posterior end. 
A retention cyst 5.5 by 4 centimeters, which projected 1.5 centi- 
meters above the surface, was present at the anterior end. 
This retention cyst was denuded of parenchymatous tissue on its 
upper surface, being covered only by a fibrous connective tissue 
capsule. At the tip of the posterior end, the capsule was adher- 
ent to a scar tissue on the kidney. This scar measures 3 by 
1.5 centimeters. 

On sectioning this kidney, an oval encapsulated abscess, 1.3 by 


282 _. The Philippine Journal of Science 1914 


1.5 centimeters, was found at the posterior end, and extended 
through the cortex into the medullary substance. Considerable 
degeneration of the parenchymatous tissue was present in the 
vicinity of this abscess. Two worms were found in the upper 
part of the ureters. Worm tracts either containing worms or 
empty were found located along the edges and between the 
papillae throughout the entire organ. These tracts were all 
similar in structure, and varied from 1 to 1.5 millimeters in 
diameter. 

The left kidney measured 15 centimeters in length, 6.5 centi- 
meters in width at its posterior end, 8 centimeters at its anterior 
end, and was 3.5 centimeters in thickness. 

In the central portion above the hilus, a scar 2.5 by 1.5 centi- 
meters was found. At the posterior end, the capsule was 
adherent to a scar 4 by 1.5 centimeters. On section, 2 worms 
were found embedded in the hilus. 

Numerous worm tracts, either containing worms or filled with 
purulent material, were found in the connective tissue located 
along the spinal column in the vicinity of the kidneys. 

Pleurisy was very evident. The lobes of the lungs were 
adherent to each other, to the costal pleura, and to the dia- 
phragm. Several worms were found free in this cavity, and 2 
were embedded in the costal pleura. 

The pericardial sac contained a large amount of blood-stained 
fluid. The heart and pericardium were covered with a thick 
feltlike fibrinous exudate, and were adherent to each other. The 
heart and pericardium presented the appearance of the marked 
traumatic pericarditis which is sometimes found in cattle. Five 
worms were found at the base of the aorta. 

’ Worms were found free and embedded in both pleural and 
peritoneal cavities. Twenty-eight were picked up which were 
lying free among the organs. 

Berkshire sow A-25, age 4 years 3 months and 21 days, died 
during the night of August 13, 1913. 

On looking over the history of this animal, the following may 
be summed up. During her life, she aborted once and gave birth 
to 32 pigs of which 14 lived. From these figures, it will be noted 
that this animal was not profitable for breeding purposes. 


AUTOPSY 


The animal was large and from external appearances in fairly good 
condition. Upon autopsy, a large amount of clotted blood and dark blood- 
stained liquid was present in the peritoneal cavity. This appearance sug- 


1) B, 8 Boynton: Kidney-worm Infestation of Swine 283 


gested that the immediate cause of death was from an internal hemorrhage. 
The source of the hemorrhage was found to be from a large aneurysm of 
the anterior mesenteric artery which had ruptured. This aneurysm is 
represented in Plate III, fig. 2. The anterior end, a, lay in close proximity 
to the posterior aorta. It measured 24 centimeters in length and 34.5 centi- 
meters in circumference around the largest portion. Near the posterior 
end there was a slight constriction, f, which may be considered as dividing 
the lesion into two aneurysms, a large anterior and a small posterior; how- 
ever, the constricted portion was not composed of normal tissue. The 
tunica adventitia was markedly roughened by outgrowths of fibrous connec- 
tive tissue and nodules. These nodules (Plate III, fig. 2, e, and fig. 3, 
a and 6), of which 11 were easily seen on the surface of the specimen, were 
composed of fibrous connective tissue. They protruded above the surface 
on an average of 0.6 centimeter, and varied from 0.5 to 0.9 centimeter in 
diameter. The longest nodule measured 3.5 centimeters in length, and 
formed a tortuous course. The majority were about 1.5 centimeters in 
length, giving them an oval appearance. These nodules either contained 
kidney worms (Plate III, fig. 3, a), in which instance a worm can be seen 
projecting from the cut end of a nodule, or they were filled with a mass 
of cheeselike material. The tunica adventitia was very thin, being made 
up of merely a superficial layer of connective tissue in places. The endo- 
thelial lining and tunica intima had entirely disappeared and their places 
were taken by a thick layer of fibrinous material varying from 0.3 to 2.6 
centimeters in thickness, which coated the entire aneurysm. This fibrinous 
material (Plate III, figs. 2, c, and 3, c) was yellowish white, firm in consis- 
tence, and tough, thus forming a protective coat over the inner surface of 
the aneurysm. 

A large parietal thrombus was present in the aneurysm (Plate III, fig. 4). 
Two kidney worms were embedded in this thrombus, b and 1b, and one worm 
was found with its body partially protruding into the cavity of the aneurysm, 
a portion of the body still remaining embedded in the wall. These facts 
show that the kidney worm is capable of working its way through the wall 
and into the lumen of a blood vessel. 

The appearance of the lesion just described suggests that by pene- 
trating the wall of an artery the kidney worm is capable of producing 
an inflammation of the muscular coats, which may cause them to lose their 
tone. As a result of the constant high blood-pressure and weakened vessel 
walls, they were stretched and unable to regain their normal size, the 
ultimate result being the formation of the aneurysm. The heavy coating 
of fibrinous coagulated material undoubtedly arose from the endaortitis, 
which may have been caused directly by the irritating effect of the worms 
upon the endothelial layer or by the extensive stretching of the weakened 
muscular walls of the blood vessel. 

The aneurysm thus formed corresponds in many ways with the aneurysm 
found in horses infested with Strongylus armatus (Sclerostoma equinum). 
In the horse, the immature worm causes the aneurysm while in kidney- 
worm infestation mature worms were found. 

In this animal, there was a generalized infestation. Worms were found 
both free and encysted in the thoracic and peritoneal cavity. Three worms 
were found in the right kidney, 2 of which were coiled in the calix and 1 
embedded in the medulla. 


284 The Philippine Journal of Science 1914 


The lesions present in these animals go further to prove that 
kidney-worm infestation becomes a generalized disease if allowed 
to run its course. They also show how detrimental the disease 
may be to a drove of breeding animals. It not only kills off the 
older breeding animals, but may cause them to abort. In some 
instances it has been noticed that sows die soon after giving birth 
to a litter of young, causing the loss of the entire litter or stunt- 
ing them to such an extent that they will never be as large animals 
as those from a healthy mother. Also, there is a possibility that 
the entire litter may become infected from the mother. 


MEDICAL TREATMENT 


Since the worms are embedded in the solid tissues and are 
surrounded by purulent débris, treatment with vermicides in- 
troduced by the mouth is absolutely unsatisfactory. Remedies 
never could be expected to reach the worms in sufficiently con- 
centrated form to have any beneficial action. Hence prevent- 
ive measures are of more than usual importance. 


PREVENTION 


Various investigators have come to the conclusion that the 
worm passes no part of its life cycle in any other animal than 
the pig. Therefore, every effort must be made to break the 
chains of succession in the reproduction of the worm by not 
allowing uninfested animals to come in contact with infested 
ones or with their excretions. 

Law * states that— 


Hogs should be excluded from all ground known to be infested, or on 
which infested hogs have been, or which receive drainage from fields, lots 
or pens occupied by other hogs; * * *. Above all the pigs should be 
kept apart from slaughter houses and streams into which they drain and 
on no account should they be allowed the offal or flesh of other pigs, 
including scraps from the kitchen, unless the material has been thoroughly 
cooked. * * * As in the case of other communicable diseases of pigs, 
the massing of these animals in large herds in contaminated localities is 
particularly dangerous. * * * Purchased pigs should only be added 
to sound herds on the basis of irrefragible evidence of the soundness of the 
herds and localities from which they come, and even then only after 
quarantine. 


The only way the disease can be controlled to any extent is 
by placing the pigs on slat floors, which allow the urine and 
feeces to drain through. In this way, the uninfested animals 
are not so apt to some in contact with infective material. 


* Veterinary Medicine, 2d ed. (1909), 5, 542. 


—-— 2s = -..—hS  U 


IX, B, 3 Boynton: Kidney-worm Infestation of Swine 285 


SUMMARY 


1. From the specimens procured from hogs by me and from 
identification of the same by H. B. Ransom, it is proved that 
the animals were infested with Stephanurus dentatus. 

2. From observations made by R. W. Newcomb and by A. S. 
Shealy on native hogs killed at the matadero in Manila and from 
identifications made by me of specimens collected by them from 
about 25 cases, it is proved that Stephanurus dentatus infesta- 
tion is prevalent among the native pigs slaughtered there. 

3. The age at which pigs usually die from infestation with 
Stephanurus dentatus, as observed by me, and the lesions pro- 
duced by this worm indicate a slow-developing, chronic disease. 

4. Since the average age of pigs that are killed for meat at 
the Manila matadero varies from six months to one and a half 
years, the disease does not have time to cause any fatal or very 
damaging lesions in the animals. Hence it is not looked upon 
as very serious in connection with meat inspection. In older 
animals, the lesions are more serious. 

5. From the several autopsies made on animals, it is con- 
cluded that kidney-worm infestation becomes a generalized in- 
stead of a localized disease of swine when allowed to run its 
course. Practically every cavity of the body may become in- 
fested. 


ACKNOWLEDGMENT 


The writer is indebted to Mr. H. B. Ransom, chief, zoological 
division, Bureau of Animal Industry, Washington, D. C., for 
identification of the parasite and other information. 

Dr. R. W. Newcomb and Dr. A. S. Shealy assisted in collecting 
information and material for study. 

Mr. L. J. Fattey, foreman on the Alabang stock farm, furnished 
the history of several animals mentioned in this paper and 
promptly notified me in cases where animals were either sick 
or dead. 


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ILLUSTRATIONS 


PLATE I 


Fic. 1. Section of liver from a pig infested with kidney worms. 

a, markedly congested lobule in the dependent portion of a lobe. 

6, a worm tract in the interstitial tissue. 

c, an accumulation of worm tracts. 

d, a marked development of interstitial tissue from the chronic 
productive inflammation which has practically replaced the 
parenchymatous tissue. 

2. A section of liver from a pig infested with kidney worms and which 
shows thrombus formation. 

a, hepatic vein. 

b, a parietal thrombus measuring 1.9 centimeters in length and 
filling approximately half of the vessel. 

c, worm tracts in the immediate vicinity of the vessel wall which 
are surrounded by connective tissue. 

d, worm tracts deep in the liver substance, replacing the paren- 
chymatous tissue with fibrous connective tissue. 

3. Kidney of a pig infested with kidney worms. 

a, large retention cyst. 

b, scars on the surface of the kidney. 

c, the mottled appearance over the surface of the kidney. 

4. Transection of a kidney from a pig infested with kidney worms. 

a, cortical portion of kidney, presenting perforations undoubtedly 
caused by post-mortem changes. 

6, medullary portion of kidney presenting the same porous appear- 
ance. 

c, kidney worms coiled up in the pelvis of the kidney. 

d, a kidney worm penetrating the medullary portion of the kidney. 

€, an accumulation of worm tracts in the pelvis of the kidney. 

f, worm tracts extending into the medullary portion of the kidney. 


PLATE II 


Fic. 1. Microphotograph of the liver from a pig infested with kidney 
worms. 
a, hemorrhagic areas around the hepatic vein, showing marked 
passive congestion. 
b, interstitial tissue somewhat thickened. 
2. Microphotograph of the kidney, from a pig infested with kidney 
worms. 
a, casts present in the convoluting and collecting tubules. 
b, formation of casts in the tubules. 
c, the breaking up of the parenchymatous cells and the extrusion 
of colloid material into the lumen of the tubules in the forma- 


tion of casts. 
287 


Ct ROLF 


288 The Philippine Journal of Science “1914 


Fig. 2 —Continued. 
d, degeneration of the parenchymatous cells; primary step in the 
formation of the casts. 

3. Highly magnified microphotograph of a section of the kidney from a 

pig infested with kidney worms. 
, parenchymatous cells, containing minute spherical droplets of 
colloidlike material. 

b, droplets of colloidlike material, bursting out of the cells and 
beginning to accumulate in the lumen of a tubule. 

c, cells, the cell bodies of which have undergone complete degener- 
ation and are filled with droplets of colloidlike materials. 

d, the nuclei of cells which have undergone degeneration and still 
retain their staining properties. 

e, the lumen of a tubule practically filled with droplets of colloid- 
like material which have not coalesced. 

f, casts formed in the tubules from the droplets Berne by the 
parenchymatous cells. It will be noted that the tubules in 
which these casts are located are practically denuded of their 
epithelium. 

4. Microphotograph of a section of a lymph gland, showing thrombus 
formation from a pig infested with kidney worms. 

a, lymphoid tissue. 

b, thickened wall of blood vessel. 

c, obturating thrombus in the artery. 

d, thickened trabecule. 


PLATE IIT 


=) 


Fic. 1. Microphotograph of a section of a lymph gland containing a worm 
tract, from a pig infested with kidney worms. 
a, worm tract filled with fibrin, cell detritus, and leucocytes. 
b, newly formed fibrous connective tissue capsule around a worm 
tract. 
c, newly formed blood vessels in the fibrous connective tissue 
capsule. 
d, a worm tract of longer duration than that of (a), containing 
fibrin, cell detritus, and leucocytes. 
e, a homogeneous zone, suggesting calcification. 
f, well-formed connective tissue capsule. 
g, an old worm-tract, containing a granular mass of material. 
h, a calcified zone. 
i, a well-formed old connective tissue capsule. 
2. An aneurysm of the anterior mesenteric artery. 
a, anterior end of the mesenteric artery in close proximity to the 
posterior aorta. 
, distal end of mesenteric artery. 
a section in the wall of the aneurysm, showing its thickness, 
most of which is composed of a coagulated fibrinous mass. 
, the area from which the hemorrhage took place. 
nodules on the surface of the aneurysm, containing either 
kidney worms or cell detritus. 
f, a constriction which divides the lesion into a large anterior 
and small posterior aneurysm. 


i a 


, 


s a 


, 


1 ade? 
b, nodules in the wall of the aneurysm. 
CG thick fibrinous coagulated material, which formed a coating 
over the entire inner surface of the aneurysm. 
d, the thin ue pee 


: cas ; of hee Fie ebus. . 
b and 1b, two kidney worms embedded in the thrombus. 


’ 


AEC ings ELF ae Bie ee 


eg 


‘ant 


BOYNTON: KIDNEY-wWoRM INFESTATION OF SWINE. | [PuHit. Journ Scr, IX, B, No. 3. 


Fig. 1. A section of the liver. Fig. 2. A section of the liver. 


Fig. 3. An infested kidney. Fig. 4. A transection of an infested kidney. 


PLATE I. KIDNEY-WORM INFESTATION OF SWINE. 


BOYNTON: KIDNEY-WORM INFESTATION OF SWINE. ] [PHit. Journ Sct., IX, B, No. 3. 


Fig. 1. A microphotograph of the liver. Fig. 2. A microphotograph of the kidney. 


Fig. 3. A microphotograph of the kidney. Fig. 4. A microphotograph of a lymph gland. 


PLATE Il. KIDNEY-WORM INFESTATION OF SWINE. 


ies ay, Pub 


Cat 


PoYNTON: KIDNEY-WORM INFESTATION OF SWINE. ] [PuHit. JourN Scr., IX, B, No. 3. 


Fig. 1. Section of a lymph gland. Fig. 3. A portion of the aneurysm. 


Fig. 2. An aneurysm cf the anterior mesenteric Fig. 4. The thrombus which was Present in the 
artery. aneurysm. 


PLATE Ill. KIDNEY-WORM INFESTATION OF SWINE. 


ANOTHER DANGEROUS JELLYFISH IN PHILIPPINE WATERS 


By S. F. Licut 
(From the Zoological Laboratory, College of Liberal Arts, 
University of the Philippines) 

During the stay of the biological survey party of the Univer- 
sity of the Philippines and the Bureau of Science in Taytay, 
Palawan, in 1913, one of the women of the party was severely 
poisoned by the sting of a jellyfish hitherto not specifically re- 
ported as dangerous. A number of specimens of this medusa 
were procured, and since returning to Manila I have been able 
to indentify it as Chiropsalmus quadrigatus Haeckel, a species 
named by Haeckel from a specimen collected at Rangoon. 
Mayer + has recently redescribed it from specimens collected by 
the United States Bureau of Fisheries ship Albatross in various 
parts of the Philippines. 

It was unfortunate that there was no doctor in the party to 
preserve an accurate and technical account of the symptoms, but 
I give the following untechnical description in the hope that it 
may be at least of general interest to medical men in the Islands. 

The stings were inflicted on the feet and legs, the long and 
delicate tentacles wrapping several times around the legs and 
breaking from the body of the medusa. These tentacles, bearing 
tens of thousands of minute nettling cells, clung to the skin, and 
wherever they touched a purplish swollen ridge appeared with- 
in a few seconds. Within a few hours, each of these ridges was 
marked by a continuous blister, and although these were opened 
several times they persisted for about a week. The red marks 
which were left wherever the tentacles had touched the skin were 
still very distinct, some four months after the sting was in- 
flicted. This bears out the statements of the people of Palawan, 
who say that these marks always persist for six months or even 
more. Swelling of the legs and feet began almost immediately, 
and persisted for from seven to eight weeks, the person stung 
not being able to walk because of this swelling. About ten 
minutes after the sting was inflicted, general muscular spasms 
ensued. Those of the respiratory muscles began after about 


*Medusae of the World (1910), 3, 516. 
291 


292 The Philippine Journal of Science 1914 


twenty minutes and were especially severe, making breathing 
very difficult at times. In about half an hour the heart action 
became very feeble, and the patient was semiconscious for a 
period of about fifteen minutes, during which time the pulse 
could be detected with difficulty. Although the patient had been 
given 3 doses of whisky, it was thought necessary to administer 
by mouth a dose of %) grain of strychnine, after which the 
heart action improved. 

The pain from the sting was immediate and severe, ane prob- 
ably because the surface affected was so great seemed general 
in the feet and legs, the patient saying that it was like the pain 
of a very severe burn. The patient was confined to bed for 
five days, and found it difficult to move about freely for several 
weeks thereafter, owing to the increase of pain and a return 
of the swelling and at first to a recurrence of the muscular 
contractions. The nervous shock was great, and the patient did 
not fully recover from it for two months or more. The imme- 
diate shock and effect of the poison was so great that, although 
the sting was inflicted in shallow water only a few meters from 
shore, the patient was only able to reach shore with the aid of 
fellow-bathers and had to be carried from the shore on a 
stretcher. Such a sting if inflicted in deep water would be a 
very serious matter aside from its after effects because of the 
immediate danger of drowning. 

The stings were treated locally at first with alcohol and vinegar 
‘ and then with a dressing made of soda and olive oil. The natives 
suggested vinegar and sugar. Later, the affected parts were 
wrapped in cloths moistened with a 2 per cent aqueous solution 
of cocaine which, however, gave no perceptible relief. 

The jellyfish which inflicted these stings belongs to the order 
Carybdeide, formerly called Cubomeduse because of the squared 
or cubical shape of the bell; the species of this order are some- 
times known as “sea wasps” because of their reputation as 
stingers. These jellyfish are very different from the more typ- 
ical meduse of the order Semaeostomez of which the small 
white form with long oral palps and slender marginal tentacles, 
common in Manila Bay at certain seasons of the year, may be 
taken as an example. For reasons stated in a recent paper on 
Philippine Scyphomedusez,”? I consider this medusa to be the 
“Chrysaora stage” of Dactylometra quinquecirrha L. Agassiz.® 
This is the common “sea nettle’ of the Atlantic coast of the 


* This Journal, Sec. D (1914), 9, 198. 
* Mayer, Ibid. (1910), 3, 585-588. 


a 


——— 


—_— a 
v 


1x,B,3 Light: Dangerous Jellyfish in Philippine Waters 293 


United States, which is able to inflict a very severe and in some 
cases dangerous sting. 

The large white or white and purple form common in Manila 
Bay during the summer months is Lobonema mayeri Light, 
which may prove to be identical with L. smithii of Mayer. 
While able to inflict a very unpleasant sting, this medusa is 
not dangerous. This species and the common black medusa, 
Catostylus purpurus Mayer, which is harmless, belong to the 
order Rhizostome. 

Dactylometra may be recognized by its long ribbonlike oral 
lappets and its numerous (24), slender, white, marginal ten- 
tacles. Lobonema may be recognized by its large size, by its 
long tapering tentaclelike marginal lappets, and by the long 
slender filaments which hang down in large numbers from the 
mouth arms. 

In Dactylometra which the Filipinos call fosforo the sting 
is inflicted by nematocyst batteries in the 4 long ribbonlike 
oral palps, in Lobonema which the Filipinos call lanterna it is 
inflicted by the nematocysts of the long filaments which arise 
from the mouth arms, and this medusa may be handled with 
impunity if care is taken not to come in contact with these fila- 
ments, while in Chiropsalmus the nematocysts are located in 
the long marginal tentacles. 

Chiropsalmus quadrigatus may be recognized by its deep 
transparent bell, about as high as thick and flattened laterally 
in 4 planes, and by its very long slender tentacles banded 
with lavender areas and arranged in 4 groups, typically 7 to 
a group, each group arising from a stiff hand-shaped projection 
of the exumbrella. The medusa reaches a diameter of 200 milli- 
meters or more, with tentacles 1.5 meters in length. It is found 
in shallow water, and is especially dangerous as its transparency 
renders it very inconspicuous and it is apt not to be noticed by 
the bather. 

The severe stings inflicted by this medusa are easily under- 
stood when the tentacles are studied under the microscope. The 
violet or lavender bands spoken of above are completely covered 
on their outer surface with thousands of closely approximated, 
slender, comparatively large, stinging thread cells or nema- 
tocysts. A nematocyst consists in general of a capsule, the outer 
end of which is extended to form a long hollow hair, which 
usually bears on its outer surface recurved hooks or spines. 
At rest, this hollow hair is introverted and lies coiled within 
the capsule. 


294 The Philippine Journal of Science 1914 


There is usually a little trigger-shaped projection of the ecto- 
derm cell in which the capsule lies. This projection is called the 
cnidocil, and when touched causes the discharge of the stinging 
hair which turns out on itself as when one blows out the in- 
turned finger of a glove. Piercing the skin while only partially 
everted, it continues the process within the tissues, at the same 
time discharging the poisonous fluid which it contains. Glaser 
and Sparrow‘ found the discharge of nematocysts to be controlled 
by changes in osmotic pressure. While the amount of the fluid 
discharged from a single nematocyst is very slight, the nema- 
tocysts are sO numerous in a small area that the aggregate 
amount of poison discharged into the tissue must be considerable. 
This fluid has never been analyzed from any of the dangerous 
medusz, and it would seem that this form which is very com- 
mon at a certain season of the year in many places in the Islands, 
particularly in Culion and in Subig Bay, would afford excellent 
opportunities for study along this line, because of its enormous 
numbers of localized nematocysts. 

Specimens of this medusa were collected by the Albatross 
at several points in Luzon and at Masbate and Mindanao. 
While with the biological survey party, I found them to be plen- 
tiful on the Sulu Sea side of Palawan and at Culion and 
Busuanga. The species is apparently widely distributed in the 
Archipelago, and bathers should be on the lookout for it. The 
natives in Palawan report numerous severe stings, some of them, 


particularly in the case of children, resulting in death, and it - 


was notable that after the appearance of the medusa in that 
region the children were almost never seen in the water. 

Old* of the United States Navy and formerly stationed at 
Canacao, Cavite, has reported several cases of severe poisoning 
by Scyphomedusz in two of which death occurred. The jellyfish 
responsible for these cases were probably Dactylometra, although 
Dr. H. M. Smith thinks they were due to Lobonema smithii 
Mayer.® I have never seen Lobonema smithii, but I have seen 
numerous cases of stings by Lobonema mayeri which, as I have 
said, may prove to be identical with Mayer’s L. smithii, the 
main differences being in the number of radial canals. Further- 
more, I have experienced these stings myself, and while by no 
means pleasant they are not dangerous. Catostylus purpurus 
Mayer and Auwrellia labiata Cham. and Eysen., the only other 


* Journ. Exp. Zool. (1909), 6, 361-382. 
° This Journal, Sec. B (1908), 3, 329. 
* Mayer, Medusae of the World (1910), 3, 690-691. 


: 
| 


1x,B,3 Light: Dangerous Jellyfish in Philippine Waters 995 


common meduse in Manila Bay, are harmless, and I have handled 
them on numerous occasions without suffering any sting whatso- 
ever. Dactylometra on the other hand is known to be very 
dangerous. Several cases of severe stings inflicted by it are 
known, and the natives, particularly the fishermen, hold it in 
great fear. While the symptoms reported by Old were quite 
different in many ways from the symptoms usually following 
the sting of Dactylometra and those in the case reported here, 
and while they may have been due to Lobonema smithii or some 
hitherto unreported medusa, it seems probable that they were 
merely the results of very severe poisoning from the sting of 
Dactylometra. 
126334——7 


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SOME SIMPLE LABORATORY APPARATUS 


By R. B. GIBSON 
(From the Department of Physiology, College of Medicine and Surgery, 
University of the Philippines) 
Three text figures 


The results of the physiological experiments, as taught in the 
laboratory, are often lost to the student because the complexity 
of the apparatus distracts him from the actual observation. 
Moreover, the use of expensive physiological apparatus in many 
experiments necessitates either demonstration work or the em- 
ployment of groups of students. I have found, particularly with 
Filipinos, that it is highly desirable from a pedagogic stand- 
point to simplify the experiments so that more of them may be 
performed and reported by each student individually. 

In developing a course along these lines, three pieces of appa- 
ratus have already proved so useful that it has seemed worth 
while to describe them. 


THE THERMOESTHESIOMETER 


This thermoesthesiometer (fig. 1) is made by inserting a wire 
nail through a small cork which is then fitted into the constricted 
end of the glass chamber of the Porter muscle warmer or finger 
plethysmograph (supplied by the Harvard Apparatus Company). 
The other opening is provided with a suitable cork. A wire 
test-tube holder forms the handle. 

For use, the glass chamber is filled with ice shavings or with 
hot water. A single charge of the ice or hot water is more than 
sufficient for locating the hot or cold points of the skin, when 
observed in the usual way. 


A VENOUS-PRESSURE APPARATUS 


The simple arrangement (fig. 2) may be used in place of the 
von Recklinghausen: and the Eyster and Hooker? apparatus for 
venous pressures in man. 

A finger of a discarded rubber surgical glove is cut off and 
drawn over a small glass filter funnel. The tip of the finger is 
carefully trimmed away as in the illustration. The funnel is 


*Arch. f. Exp. Path. u. Pharm. (1906), 55, 470. 
* Bull. Johns Hopkins Hosp. (1908), 19, 274. 
ake 297 


298 The Philippine Journal of Science 1914 


connected by tubing through a T to a manometer and a rubber 
bulb. 

To make an observation, the skin over the selected vein of the 
forearm is swabbed with glycerol, the funnel is inverted upon 
this area and is held lightly in place. A slight positive pressure 
within the funnel seals the rubber flange against the skin and 
the funnel itself may be raised off the vein. The collapse of the 
vein may be observed through the glass sides of the funnel. 
Hinged blocks for the pressure bulb increase the accuracy of the 
results. 


Fic. 1. The thermoesthesiometer. 


A SIMPLE TIMING SYSTEM 


While most laboratories are satisfactorily equipped with a 
time-marking system for class purposes, no very satisfactory 
instruments’ of this kind are commercially available. In many 
laboratories one finds timing systems which have been devised 
and made in the school itself. The excellent electric clock which 
is part of the equipment of the majority of American laboratories 
is too delicate an instrument for continuous use or for supplying 
a number of work tables. 

The apparatus here described (fig. 3) can be set up with a few 
hours’ work, and has given excellent service for twelve laboratory 
benches. 


— 


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. eke Rls te 
wr 


IX, B, 3 


Gibson: Simple Laboratory Apparatus 


Fic. 2. Venous-pressure apparatus. 


Fic. 3. Arrangement of wall clock for timing purposes. 


299 


300 The Philippine Journal of Science 1914 


A watch spring is inserted in the woodwork of an eight-day 
wall clock, with a pendulum of a half-second period. The 
arrangement is such that a contact is made each second at the 
end of the swing between the extremity of the pendulum rod 
and the spring. Parallel wires lead from the works of the 
clock and from the spring through binding posts in pairs on 
the several work benches. Six large dry cells furnish the neces- 
sary electricity to operate the signal magnets which may be 
connected to the binding posts on the tables. 

With this apparatus, electricity is not being used up except 
when the circuit is closed by the insertion of a signal magnet. 
The timer is always ready when it is necessary to make a record. 
Two or twelve signal magnets may be used simultaneously, and 
may be inserted on any bench without reference to connections 
on other tables. The time in seconds will be represented on the 
record as a Single vertical projection from the base line. 

The apparatus requires no attention except winding the clock 
once every week. Occasionally the contact point on the pendu- 
lum should be polished with emery and a little mercury rubbed 
into the brass. The spring should be kept free from rust. 


_ ILLUSTRATIONS 


TEXT FIGURES 


hermoesthesiometer. 
nc ressure apparatus. 
Arrangement of wall clock for timing purposes. 


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venir OHTA TY 


REVIEWS 


The | Practice | of | Medicine | a text-book for practitioners and students | 
with special reference to diag- | nosis and treatment | by | James 
Tyson, M. D., LL. D. | [4 lines] | and Mr. Howard Fussell, M. D. | 
[4 lines] | Sixth edition, revised and rewritten | with six plates | and 
179 other illustrations | Philadelphia | P. Blakiston’s Son & Co. | 1012 
Walnut Street | 1913. 

The present revised edition of Tyson’s Practice of Medicine 
is quite up-to-date and it contains many new subjects. The 
most important among these are: Diseases of the Pituitary 
Gland, Diverticulitis, Diseases of the Thymus Gland, Hypothy- 
roidism and Hyperthyroidism, Osteopsathyrosis, Oxicephaly, and 
many others. It is, however, to be regretted that the so-called 
Tropical Diseases have not received the proper attention that 
they deserve in their revision. The description of some of them 
is very unsatisfactory and is not consistent with the modern 
views that we have of them at present. With this minor ex- 
ception there is no doubt that this book on Practice of Medicine 
deserves to be highly recommended for its worth, and it will 
continue to be one of the favorite textbooks among medical 
students, on account of the preciseness and clearness with which 
it deals with the whole subject of internal medicine. 

A. G. SISON. 

Pathogenic | Micro-organisms | a text-book of | microbiology for physicians | 
and students of medicine | by | Ward J. MacNeal, Ph. D., M. D., | 
professor of pathology and bacteriology in the New York | post- 
graduate medical school and hospital, New York | (based upon Wil- 
liams’ Bacteriology) | with 213 illustrations | Philadelphia | P. Blak- 
iston’s Son & Co. | 1012 Walnut Street | 1914 | Cloth, pp. i—xxii+ 
1—462. Price $2.25. 

This volume should more properly be termed a manual than 
a textbook, and like many of its class it attempts too much, 
rather than too little. In a hasty glance through the book the 
following criticisms may be mentioned. That portion of Chap- 
ter I dealing with the development and principles of the micro- 
scope might well have been omitted. In Chapter II under test- 
ing disinfectants no mention is made of the U. S. Hygienic 
Laboratory method: a very important omission in the opinion 
of the reviewer. The use of Dieudonné’s medium is ignored 
under the bacteriological diagnosis of cholera. Quite a few typo- 

303 


204 The Philippine Journal of Science 1914 


graphical errors occur throughout the text, but the book on 
the whole will undoubtedly fill the purpose for which it was 
written. 
J. A. JOHNSTON. 
Man a Machine | by | Julien Offray de la Mettrie | French-English | in- 
cluding Frederick the Great’s | “Eulogy” on la Mettrie and ex- | tracts 
from la Mettrie’s “the | natural history of the soul’ | philosophical 
and historical notes | by | Gertrude Carman Bussey | M. A., Wellesley 
College | Chicago | The Open Court Publishing Co. | 1912 | Cloth, 
pp. 1-216. Frontispiece. 

The essays which outline La Mettrie’s metaphysical doctrines 
are chiefiy of historical interest. In his argument in favor of 
materialism, he draws on many experimental observations, some 
original, to develop the physiological analogy between man and 
the lower animals. In spite of the deficient physiological knowl- 
edge of the time (1748), La Mettrie’s arguments are rational, 
and many parts of the essays read like mechanistic papers of 
the present day. 

R. B. GIBSON. 


Practical | Bacteriology, Blood Work | and | Animal Parasitology | including | 
bacteriological keys, zodlogical tables | and explanatory clinical note3 | 
by | E. R. Stitt, A. B., Ph. G., M. D. | medical inspector, U. S. Navy; 
graduate, London school of tropical medicine; head | [etc. 6 lines] | 
Third edition, revised and enlarged | with 4 plates and 106 other 
illustrations containing 513 figures | Philadelphia | P. Blakiston’s Son 
& Co. | 1012 Walnut Street | 1913 | Cloth, pp. i-xv+1-408. Price, 
$1.50. 


While the third edition of this manual is enlarged and brought 
up to date in a general way, the most notable changes from the 
second edition are found in the section on animal parasitology. 
Many of the illustrations in this section are new and they 
are excellent. This book could not replace the larger textbooks 
upon the various subjects with which it deals, but like the 
preceding editions it will doubtless be extremely useful not 
only to the busy practitioner but to the student as well. 

DAVID G. WILLETS. 
The American | Illustrated | Medical Dictionary | a new and complete dic- 
tionary of | the terms used in medicine, surgery, | dentistry, phar- 
macy, chemistry, nurs- | ing, veterinary science, biology, medi- | cal 
biography, etc., with the | pronunciation, derivation, and definition | 
including much collateral information | of an encyclopedic character | 
by W. A. Newman Dorland, A. M., M. D. | member of committee on 
nomenclature, [etc. 4 lines] | together with new and elaborate tables 
of arteries, muscles, [etc. 4 lines] | seventh edition, revised and 


enlarged | Philadelphia and London | W. B. Saunders Company | 
1913 | Pp. 1107, flexible leather, $4.50. 


IX, B, 3 Reviews 805 


This new edition of “Dorland’” is an excellent and convenient 
medical dictionary brought well up-to-date by the introduction 
of over 5,000 new terms, including advances in all branches of 


medicine. 
Bs C.-C: 


An Essay on | Hasheesh | including | observations and experiments | by | 
Victor Robinson | contributing editor, Medical Review of Reviews | 
[ete. 3 lines] | Medical Review of Reviews | Two hundred and six 
Broadway | New York | 1912 | Cloth, pp. 1-83. Price $0.50. 


Cardio-vascular Diseases | recent advances in their anatomy, physi- | ology, 
pathology, diagnosis and | treatment | by | Thomas E. Satterthwaite, 
A. B., M. D., LL. D., Se. D. | Consulting Physician Post-Graduate, 
Manhattan State, Orthopedic, | Babies’, Champlain Valley Hospitals 
and North Eastern Dispen- | sary; etc. [11 lines] | [motto] | Lemcke 
and Buechner | 32 West 27th Street New York City | no date. Copy- 
righted 1913. Cloth, pp. 1-166, 80 text figures. 


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THE BATAN DIALECT AS A MEMBER 
OF THE PHILIPPINE GROUP 
OF LANGUAGES 
By Orto Scuerer 
and 
“EP” AND ‘‘V” IN. PHILIPPINE 
LANGUAGES 
By CarLos Evexerr Conant 


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THE SUBANUNS OF SINDANGAN BAY 
By Emenxson B, Cristie 


Order, No. 410. Paper, 121 pages, 1 
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Sindangan Bay is situated on the north- 
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The 29 plates illustrate the Subanuns at 
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altars, and impleménts; and the people 
themselves. 


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By Nasees M. SALEESY 


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In the preparation ef his manusoript Ue 
The History of Sulu, Doctor Saleeby spent 
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Moros in the Philippines from the 
times to the American occupation, 


| ' 


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tion to the subject; it is not a mere com- 

pilation, for the author was in the field and 

understands the conditions of tome he 

writes, ‘ 

A MANUAL OF P. PINE SILE 
OUL' 


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plates, $0.75, postpaid. 

In A args Be Pai voing Silk Culture 
are presented the results of sin rel ; 
gether with « deseription ie Bs c 
pine race. , 


THE PHILIPPINE 


JOURNAL OF SCIENCE 


B. TROPICAL MEDICINE 


VoL. IX AUGUST, 1914 No. 4 


THE PIPETTE METHOD IN THE ISOLATION OF SINGLE MICRO- 
ORGANISMS AND IN THE INOCULATION OF SUBSTANCES 
INTO LIVING CELLS 


WITH A TECHNIQUE FOR DISSECTION, STAINING, AND OTHER PROCESSES 
CARRIED OUT UNDER THE HIGHER POWERS OF THE MICROSCOPE * 


By MARSHALL A. BARBER 


(From the Biological Laboratory, Bureau of Science, Manila, P. I.) 


Two plates and 19 text figures 


CONTENTS 
INTRODUCTION. Preparation of organisms for iso- 
ISOLATION OF MICROORGANISMS. lation. j 
General principle. Nutrient media. 
Method I. Tilumination. 
Apparatus. Pipettes. 
Technique of isolation. Gas for the microburner. 
Method II. SPECIAL APPLICATIONS OF THE PI- 
Method III. PETTE METHOD. 
CULTIVATION OF THE ISOLATED OR-| Isolation of organisms from quan- 
GANISM. tities of water larger than hang- 
Cultivation in situ on the cover ing drops. 
glass. Fixation and staining. 
Cultivation ina medium apart from! Serological tests. 
the cover glass. Experiments on chemiotaxis. 
Inoculation into animals. Dilutions. 
DETAILED DESCRIPTION OF CERTAIN Warm box. 
STEPS IN THE ISOLATION METH-| Dissection. 


ops. The pipette and isolating chamber 
Moisture. ‘ used as separate units. 
Danger of contamination. Inoculation into living cells. 


* Received for publication, December 12, 1913. 
128448 307 


308 The Philippine Journal of Science 1914 


INTRODUCTION 


Since the classical work of Hansen in isolating single cells of 
yeast plants, various methods of isolating single organisms under 
microscopical control have been described. In the method of 
Lindner, employed chiefly in the isolation of yeast cells or rela- 
tively larger organisms, droplets of a highly diluted culture are 
placed on a cover glass with a fine, sterile drawing pen. The 
preparation is placed over a moist chamber and examined for 
a droplet containing a single organism. In the well-known 
method of Burri,? a dilution of the organisms is made in India 
ink. With a sterile drawing pen, fine droplets are placed on 
solidified gelatin, and a cover glass is placed over them. The 
contrast between the organism and the dark background makes 
it possible to distinguish the organisms clearly and to find a 
droplet which contains but a single one. The organism is left 
to form a colony in situ, or the cover, with the organism clinging 
to it, is transferred to any desired medium. In the method of 
Schouten,* the single organism is picked out from a dilution by 
means of a very fine glass loop in a moist chamber and under 
microscopic control. 

The method described here is founded on a principle wholly 
different from any of those described above. No preliminary 
dilution of the micro6rganisms is necessary, except such as may 
be carried out under the microscope at the moment of isolation, 
and this dilution is required only in relatively dense suspensions. 
I first used the method in 1902 in connection with the selection 
of yeast plants. The description of the method was first pub- 
lished in 19044 and more fully in 1907.5 It was, so far as my 
knowledge extends, the first method of isolating organisms under 
microscopic control described since that of Lindner. Since the 
first publications of the technique, various modifications and im- 
provements have been described in later papers. The chief aim 
of this article is to bring together these various descriptions and 
to add considerable material not before published. 

The method has been introduced into a number of laboratories, 
and it is hoped that this description may lead to its wider use. 
It is believed that the method has a universality of application 
which will make it of use not only to bacteriologists, but also 
to workers in all departments of microscopy. Single bacteria 


*Das Tuscheverfahren. Jena. Gustav Fischer (1909). 
*Zeitschr. f. wiss. Mikros. (1905), 22, 10. 

‘Journ. Kans. Med. Soc. (1904), November. 

* Sct. Bull., Kansas Univ. (1907), 4, 8. 


ee ee ee eee ee eee Oe 


el ae 


IX, B, 4 Barber: The Pipette Method 309 


of any degree of motility or of any size visible under an ordinary 
oil-immersion lens, yeast cells, spores of fungi, alge, protozoans, 
blood corpuscles, or other histological elements may be isolated. 
A cell may be selected from a pure culture or from a myriad of 
other organisms larger or smaller than itself. Organisms may 
be selected from cultures or from the natural sources, and the 
whole process may be carried out in any fluid desired. The 
isolated organism may be cultivated in situ, transferred to any 
medium, or inoculated into an animal. Microdrganisms, stains, 
fixatives, or other chemical substances may be injected into the 
protoplasm or vacuoles of living cells. Microscopic plants, 
animals, or histological elements may be dissected or stained 
under the higher powers of the microscope. 

The isolation of organisms and some of the other applications 
of the technique may be carried out with the aid of only the 
ordinary apparatus of a biological laboratory. 

Since the earlier publications, I have had much experience 
in teaching the method to others. The difficulties of the 
technique are certainly not insurmountable, since several learners 
have, under my direction, succeeded in making pipettes 
and isolating organisms after less than an hour’s practice. The 
results of this experience in teaching have shown me some of 
the chief difficulties of the beginner, and it is hoped that the 
following description will make the technique easy to acquire 
without the assistance of personal supervision. 

In some cases details have been given which may seem super- 
fluous to many workers, but it was thought better to err on 
the side of over description than to risk leaving any point 


. obscure. 


The method of isolation of microdrganisms described here 
is, of course, not recommended as an entire substitute for plate 
methods in any routine work. In some cases the pipette method 
may be conveniently used as such, but the chief aim of ,this 
technique is to amplify the plate method and to carry out some 
isolations where the plate method is not applicable. 


ISOLATION OF MICROORGANISMS 
GENERAL PRINCIPLE 


The principle of the method, in brief, consists in the separation 
of the single organism by means of a very fine-pointed, capillary 
pipette of glass. The isolation is carried out in hanging drops 
on the underside of a large cover glass which is placed over a 
moist chamber. The organism to be isolated is touched with 


310 The Philippine Journal of Science 1914 


the tip of the pipette, into which it enters by capillarity. A 
sterile portion of the cover is brought over the tip, and the 
organism is discharged on it by air pressure through a rubber 
tube held in the mouth of the operator. The whole process is 
carried out under the microscope, under the highest powers if 
desired. 

The pipette is the most essential part of the apparatus, and 
the governing of its movements is a vital part of the technique. 
There are several ways of holding and manipulating the pipette, 
and for the sake of convenience each will be described as a 
separate method. 

METHOD I 


In this method the pipette is manipulated by means of a 
special holder clamped directly to the stage of the microscope 
or to a metal plate fastened to the underside of the stage. 


APPARATUS 


The apparatus needed are the following: 

1. A compound microscope furnished with a mechanical stage, 
preferably one allowing wide movement in both directions. 

2. The pipette holder, preferably the type with three move- 
ments.°® 

3. A specially constructed moist chamber. 

4, Large cover glasses to fit the top of the chamber. 

5. A piece of rubber tubing about 70 centimeters long and 
about 6 millimeters in diameter. 

6. Pieces of glass tubing, 15 to 20 centimeters in length, about 
4 to 5 millimeters in diameter, and with a lumen about 3 milli- 
meters in diameter. e 

7. A microburner with a very small flame. 

Some parts of the above apparatus are here described in 
detail: 

There are two types of pipette holder: the one having move- 
ments, accurately governed by screws, in three directions of 
space; the other having but two such movements mechanically 
governed, the movements to the right or left of the observer 
being accomplished by sliding the pipette in a groove with the 
fingers. The three-movement holder is preferable, as the ad- 
ditional movement makes the technique easier for the learner, 
saves time in adjusting the pipette, and enables the worker to 


° Pipette holders may be obtained from the University of Kansas, 
Lawrence, Kansas, U. S. A 


IX, B, 4 Barber: The Pipette Method OLE 


carry out more successfully certain special applications of the 
technique to be described below. 

The three-movement pipette holder containing a pipette and 
clamped in position on the microscope is illustrated in figs. 1, 2, 
and 15, showing the front, that is, the side facing the observer, 
the top, and the back, respectively. 

The adjustment wd, governed by the screw s, allows an up- 
and-down movement. The adjustment rl, governed by the screw 
s’, allows a movement to the right or left, and the adjustment 
tf and the screw s’”’ (fig. 15) allow a movement to and from 
the observer. The to-and-from movement carries the two other 
adjustments with it, and the up-and-down adjustment carries 
the in-and-out. The holder is fastened by the clamp cl to 
the metal plate pa or pb, which is screwed to the stage of the 
microscope. The pipette is held in the groove g (fig. 2) in the 
side of the adjustment vl, and is fixed by the set screw ss, 
which moves the thin plate tp. A two-movement holder is 
shown in fig. 9 and in fig. 15, B. 

The small tightening screws on the holder should be so ad- 
justed that the up-and-down movement can be easily manipulated 
with one finger only. This will allow a rapid lowering of the 
pipette. 

The holder may be clamped directly on the stage of the 
microscope, but in most types of microscope the pipette is 
brought to a more convenient level and more working room 
allowed if a metal plate (see pb, p, and pa, figs. 1, 13, and 15) 
is screwed on the bottom of the stage of the microscope. This 
may be attached with thumb screws, so that it can be easily 
taken off or put on. A convenient form is shown in 9, fig. 13. 
This type allows the holder to be set at either of two different 
levels and at two different distances from the stage. In some 
processes it is necessary to move the mechanical stage far to 
the left. In this position the isolating chamber is apt to in- 
terfere with the pipette holder, so it is well to have some arrange- 
ment for clamping the holder 1 or 2 centimeters from the stage. 
Either type p or type pa, fig. 13, will permit this movement. 
Type pa may be attached when two holders are to be used or 
when certain special processes are to be carried out. Another 
form of plate is shown in pb (fig. 1). 

As a temporary expedient, the pipette holder may be attached 

_to a flat piece of wood placed under the stage. The wood is 
shaped like the stage, perforated with an opening for the con- 
denser, and provided with a shelf on the left for attaching the 


312 The Philippine Journal of Science 1914 


i 


<— 


q | A 
i Ag 


ie 


TEIN 


Ny 
y 
Y 


Jo TU Cc 


Fic. 1. Pipette holder containing a pipette (pip). cl, clamp by which the holder is fastened to the metal 
plate pb; ud, up-and-down adjustment governed by screw s; rl, right-and-left adjustment governed by 
screw s’; g, groove in which the pipette is held by the plate tp; ic, isolating chamber; H, hood of 
pasteboard for protecting the end of the isolation chamber. 


Sena 


SRT | 
SEES 


VL 
© 


Fic. 2. Top of pipette holder. rl, right-and-left adjustment 
governed by screw s’; g, groove in which the pipette is 
held by plate tp and set screw ss. 


| 1X, B, 4 Barber: The Pipette Method 313 


pipette holder. It may be fastened to the stage by means of 
small clamps. 

The moist chamber (fig. 3) is made by fastening strips of 
glass to a slide with Canada balsam or any convenient cement. 
It is well to cement an additional strip on the slide at the open 
end s. This serves to hold water in the bottom of the chamber 
and to strengthen the whole apparatus. Pieces of slides may be 
used in making the chamber, but it is better to have glass of 
somewhat greater thickness—about 1.5 millimeters. A very 
convenient size for the moist chamber is 70 millimeters long, 
35 wide, and 28 high. If the mechanical stage in use will not 
admit so broad a chamber, one of the width of an ordinary 
slide may be used. The narrow chamber is more readily kept 
moist, the narrower cover glass suited to it is more easily cleaned, 
and the preparations grown on this cover glass are more easily 
stained and mounted than is the case with the larger cover glass. 
However, the broader chamber gives much more working room, 
and for most routine work will 
be found more convenient. It 
is advisable to make two or three 
moist chambers at a time. One 
narrow and two broad ones will ——————————————————— 
give a good working outfit. ic. 3. Isolating chamber. p, lining of 
Temporary isolation chambers 7 Nc la ar a aa 
may be made of wood or strong 
pasteboard. A convenient way is to modify a wood or paste- 
board box, cement it to a slide, and make it partially water- 
proof at the bottom with Canada balsam or any convenient 
substance. 

The height recommended is chosen because it gives working 
room for the pipette without being so far from the stage as 
seriously to diminish the light. It will be found very advan- 
tageous to replace the ordinary substage condenser with one 
which will focus the light at a point somewhat above the stage. 
This is especially adapted for work with the oil-immersion 
lens or other high powers. The moist chamber is lined on the 
sides and end with filter paper, p (fig. 3), in order to furnish 
a larger moist surface. The upper edges of the isolating 
chamber should be even and smooth, so that the cover glass 
may be easily sealed on them. 

The cover glass should be large enough to seal well to the 
sides of the chamber and to reach to the ends. It should be 
of such thickness as to permit safe handling, while not so thick 


314 The Philippine Journal of Science 1914 


as to prevent the use of the immersion lens. Thickness 2, as 
quoted in American catalogues, suits the purpose well. 

The preparation of the cover glass is a matter of great im- 
portance to the success of the technique. One should clean the 
cover carefully and then apply just enough vaseline or fat to 
prevent the small hanging drops from running together. A 
good method is to smear vaseline over a number of well-cleaned 
covers and to place them in some convenient receptacle free 
from dust. When the covers are to be used, they are freed from 
the excess of vaseline by means of soap and water, cleaned 
carefully with a dry cloth, then heated enough to soften the 
vaseline, and rubbed again while still warm. They may then 
be slightly moistened by the breath and rubbed again with a 
fresh clean cloth. The aim is to remove as much vaseline as 
possible without the use of excessive heat or any fat-dissolving 
reagents other than soap. If an excess of vaseline is left on 
the cover, small particles will appear in the droplets and may 
be mistaken for bacteria. If all is removed, the droplets will 
run together. Fine droplets placed very closely together will 
remain separate for months on a properly prepared cover. For 
the beginner, especially, success or failure may depend on the 
condition of the cover glass. Cover glasses may be prepared 
and kept in stock for use, but it is well to rub them afresh with 
a clean, dry cloth just before sterilizing. 

As to the glass tubing, the size recommended above is the 
best, but considerable latitude is allowable. Soft glass is, on the 
whole, preferable, as it is more easily worked. The fine tip 
at the end of the capillary is more easily made with hard glass, 
but the preliminary drawing out of the capillary often requires 
a blast flame, which is not always convenient to the working 
table. 

To make a serviceable microburner, one has only to bend a 
glass tube to the form shown in fig. 5, 6, to heat one end, and 
flatten it so as to form a narrow aperture. In the form shown 
in the illustration, a clamp is fastened to the back of a wooden 
block for convenience in adjusting the flow of gas. This block 
may be dispensed with and the tube kept upright by the clamp 
alone. The tip of the burner should be at a height of about 
6 centimeters above the surface of the table. 


TECHNIQUE OF ISOLATION 


It is recommended that the beginner in this technique follow 
strictly the directions given below and in the order given: 
1. Clamp the pipette holder firmly to the left side of the 


IX, B, 4 Barber: The Pipette Method 815 


stage in such a position that the groove g (fig. 1) will be nearly 
opposite the center of the condenser. 

2. Smear the upper edges of the moist chamber thickly with 
vaseline, and add enough water to cover the bottom and to 
saturate the filter-paper lining. If the moist chamber has been 
exposed to dust, it is well to flame it lightly before adding the 
vaseline or water. 

3. Sterilize the cover glass, prepared as described above, over 
a diffused heat. The flame of a gas stove or the top of the 
chimney of a Welsbach light gives a suitable heat. Avoid 
heating so much as to burn off the vaseline film. 

4. Place the cover on the moist chamber, and press it down 
so as to seal it at the edges. Mark the upper surface as 
shown in fig. 4, using India ink or a glass pencil.’ 

5. With a platinum loop or, better, with a pipette bent at the 
tip, place drops of sterile broth 
on the underside of the cover, 
of somewhat the size and ar- 
rangement shown in fig. 4. The 
bacteria may be placed in drop 
a. This end of the cover is 
placed next to the open end of 
the chamber. 

6. Place the chamber on the Fic. 4. Cover glass marked with cross lines 


stage of the microscope and of India ink, and supplied with hanging 
x i drops of sterile fluid. a, drop to which 
focus with the low power on the bacteria to be isolated are added. 


the center of the free edge of 

the cover. The edge should be approximately at the center 
of the field. Lower the objective within 2 or 3 millimeters of 
the cover glass. It is well to protect the open end of the moist 
chamber temporarily with a piece of moistened filter paper. 

7. Make the capillary pipette as follows: 

(a) Lower the flame of the microburner to a narrow blue 
flame not over 2 millimeters high. The smallest flame that 
will remain lighted should be used, and the working table should 
be free from drafts of air. 

(6) In an ordinary Bunsen flame draw out one end of a 
piece of glass tubing into a straight capillary about 0.5 
millimeter in outside diameter. 


"Fine straight lines may be made by dipping a fine capillary tube or rod 
into India ink or asphalt cement spread on a glass slide. The smeared 
capillary is laid on top of the cover in the position desired, and is then re- 
moved. India-ink lines have the advantage of being insoluble in immersion 
oil. 


316 The Philippine Journal of Science 1914 


(c) Hold the shank of the pipette in the right hand, and with 
a pair of fine forceps held in the left grasp the capillary at 
a point about 6 centimeters from the shank. The outer sides 
of both hands should rest on the table. Bring the portion of the 
capillary next to the forceps over the flame and at right angles 
to it, then lower it to a point above, not in, the flame (fig. 5). 

(d) Pull gently with the forceps, and when the glass begins” 
to soften lift it slowly from the flame and pull with the forceps 
slightly more than at first, but not too strongly. The hands 
should remain on the table during the process and the pulling 


Fic. 5. Method of making the capillary pipette. 6, microburner. 


and lifting done by turning them slightly outward. The 
capillary will separate with a slight tug—a feeling much like 
that experienced when a taut thread, held in the fingers, is 
parted in a small flame. If the point is properly made, it will 
appear as shown in b (fig. 6). 

This capillary has sufficient rigidity, and it comes to a very 
fine point. The tip is closed, but the lumen extends to the 
very end. 

It is evident that everything depends upon the amount of 
heat used and the timing of the pull and that these must vary 
slightly with the height of the flame and the diameter of the 


IX, B, 4 Barber: The Pipette Method S317 


capillary. With a little experience, one can usually tell when a 
proper point is made by the peculiar feeling described above, but, 
if desired, the point may be inspected under the low power of 
the microscope or with a good hand lens. If too little heat is 
used and the pull made too suddenly, the capillary may part 
with a snap and the tip may have an opening too large for use. 
However, if a capillary parts with a snap and is seen to end 
in a very fine point, it is well to turn up the tip (see paragraph 
e) and place the pipette in the holder for inspection under 
the microscope. Sometimes excellent pipettes, with a small 
polished opening, are obtained when the glass, in a half-molten 
condition, parts with a gentle snap, ¢ (fig. 6). 


Fic. 6. a, Completed pipette; b, c, and d, various sorts of points. 


If too much heat is used, the capillary is apt to be drawn 
into a long, hair-like ending, d (fig. 6). This is especially apt 
to be the case when the capillary is pulled with the right hand 
instead of with the forceps. Such points may be made service- 
able by clipping off the end with sterile scissors or forceps or by 
placing the end on a slide and cutting off a portion with a scalpel. 
For the best work, however, the form b (fig. 6) will be found 
preferable to the others. 

If the learner has continued difficulty in obtaining a suitable 
point, it is well to try hard glass. The capillaries are drawn 
out in the blast flame, and may be made of slightly varying 
diameters. The flame of the microburner should be slightly 
higher than for soft glass. A number of capillaries may be 


318 The Philippine Journal of Science 1914 


prepared, sealed at the tips, and kept on the laboratory table 
ready for use. 

(e) After a suitable point is made, the end of the capillary 
is turned up at right angles, as shown in a (fig. 6). This is 
done by holding the part of the capillary just back of the point 
above the small flame and lifting the point with the tip of the 
forceps or with a needle. Care must be taken to avoid fusing 
the glass and thus closing the lumen of the capillary. Not more 
than 1 centimeter should be turned, as a greater length will be 
found awkward to manipulate in the moist chamber. If a good 
point is made, it is not necessary to turn the end at exactly right 
angles. 

8. After the pipette is finished, attach the rubber tube firmly 
to the end of the shank and set the pipette in the groove of the 
holder. Slip the pipette inward, pushing it with the left hand 
and governing the set screw with the right, until the tip is 
nearly under the low power of the microscope and is turned in 
a vertical position. It should be far enough beneath the cover 
glass to avoid danger of contamination. Set the screw so as 
to hold the pipette firmly. The exact adjustment of the tip 
under the center of the low power is attained as follows: Sight 
along the edge of the cover glass and move the right-and-left 
adjustment of the holder until the tip is in line with the edge. 
Then, looking in a direction at right angles to the first, bring 
the tip to the center of the lens by moving the to-and-from ad- 
justment of the holder. Or one may make the second adjust- 
ment by sighting down the side of the tube of the microscope. 
If the tip is not now in view, it can quickly be found by moving 
some one of the adjustments of the holder. Focus the low power 
on the tip, and adjust it to the very center of the field. If desired, 
the pipette may be adjusted under the low power before the 
isolation chamber is placed on the stage. The mechanical stage 
is then moved as far as possible to the right and the chamber 
put in. One may also remove the eyepiece, turn the objective 
aside or remove it altogether, and adjust the tip below the empty 
tube. If it is adjusted to the center of the field, it will be in 
view on replacing the eyepiece and objective. The first de- 
scribed method will generally be found preferable. 

9. Lower the tip safely below the level of the hanging drops, 
focus on the under surface of the cover, and with the mechanical 
stage bring the edge of one of the drops of sterile broth into 
the field. Lower the objective until the tip is in view, then 
slowly raise the objective and tip together until the tip comes 


is 


IX, B, 4 Barber: The Pipette Method 319 


‘into contact with the cover just outside of the drop. These 


precautions are taken to avoid crushing the tip against the 
cover glass. 

Sometimes it may be difficult to see the tip at any distance 
under the cover glass on account of the refraction of light due 
to the hanging drops. In this case, focus the low power 2 or 
3 millimeters below the cover and bring the point up very slowly. 
It will usually come into view when it approaches the cover, 
especially if it is first brought up in a deep hanging drop. With 
the tip held gently against the cover, move the mechanical stage 
slightly and break off a very small portion of the point. Immerse 
the tip in the droplet of broth for a few seconds in order that it 
may become supplied with liquid. This process may be hastened 
by suction on the rubber tube, which during this part of the 
process is held in the mouth of the operator. 

If the fields of the high and low powers of the microscope 
coincide, the tip should now be brought to the very center of 
the low-power field; if not, it should be brought to a point, pre- 
viously determined, which is near the center of the high-power 
field. It is often helpful to use the micrometer eyepiece in locat- 
ing this point, especially in changing from the low-power to the 
oil-immersion lens. Now bring the tip into contact with the 
cover glass near the edge of the broth drop, and blow out a very 
small drop with the rubber tube; then, before the liquid reénters 
by capillarity, lower the point slightly. If no drop can be blown 
out, immerse the tip in the broth again and draw in more 
fluid. If a very small opening is left, it may be necessary to 
enlarge it somewhat and refill from the sterile drop before it 
is possible to discharge a droplet. The cover glass should have 
a film of moisture on it so that the liquid in the pipette will be 
in contact with liquid on the cover. It is very difficult to dis- 
charge liquid from a fine point to a dry cover. If necessary, 
the film of moisture on the cover can be supplied by placing a 
little slightly warmed water in the bottom of the isolating cham- 
ber. A little distilled water may be taken into a medicine 
dropper, heated slightly over the flame, and added gently to the 
liquid in the bottom of the chamber. The moist air, rising, forms 
a film on the cover (Plate I, fig. 1). 

With the tip just under the broth droplet, change to the high 


power. The droplet should be in the field, and when the tip - 


is raised it will come into view. It may now be easily adjusted 
to the center of the high-power field. It is obvious that care © 
must be taken not to move the mechanical stage or the cover 
glass during the change to the high power. 


320 The Philippine Journal of Science 1914 


If the tip is already perforated, one has only to fill the pipette 
with a small quantity of broth, blow out a droplet, and, with 
this as a guide, find the tip with the high power. The advantage 
of the closed point is that it allows one to gauge the size of 
the opening to suit the organism to be isolated. The breaking 
off of the point can be more accurately done under the high 
power, and one can easily find the unbroken point by immersing 
it in the broth drop near its edge, and with the edge as a guide 
locate it under the high power. For one unaccustomed to the 
technique, however, the first-described method of breaking with 
the low power and making a broth droplet may be found easier; 
the tip can usually be safely broken off under the low power. 

For ordinary isolation of bacteria, considerable variation in 
the size of the opening is allowable—about 2 to 5 microns will 
be found suitable for Bacteriwm coli, for example. If too large, 
say over 15 microns, the difficulty of isolation will be much 
greater. If too small, it will be found difficult to blow out the 
broth or to introduce larger bacteria. If the tip has a sealed 
blunt point, it is sometimes very difficult to break it open; it is 
usually best to make a new one at once. 

10. After the tip is located at the center of the high-power 
field, lower it safely below the level of any hanging drop and 
bring the hanging drop containing the bacteria into the center 
of the field, preferably at its edge. If a particular bacterium is 
to be isolated, bring it to the center of the field and cautiously 
raise the tip of the pipette until it is just below the surface of 
the liquid. The tip may be seen as a shadow immediately below 
the drop. Now, moving the finger along the up-and-down ad- 
justment of the holder, bring the tip into the drop near the 
bacterium, then lower it instantly. The bacterium usually enters 
the pipette by capillarity. Then move the hanging drop out 
of the field and raise the pipette into contact with a sterile part of 
the cover glass covered with fine droplets of condensed moisture. 
Blow out a very small drop. If the bacterium does not appear, 
move the stage slightly and blow out a second or, if necessary, 
a third or fourth drop. The droplets should be very small, and 
should be made near the edge of a hanging drop on one of the 
lines on the cover glass so that they may be easily located. If 
the bacterium sought comes out with one or more bacteria, move 
a field or so away, discharge liquid from the pipette until it 
comes out free from bacteria, return to the droplet, and if neces- 
sary dilute with a small quantity of broth. By repeating the 
selection from the diluted droplet, one can usually isolate the 
bacterium at once. 


IX, B, 4 Barber: The Pipette Method 321 


If there is difficulty in making a small drop, it is usually because 
the cover is too dry or the pipette opening too large, especially 
if large with very irregular edges. It is often possible to gauge 
the size of the droplets by raising the pipette up and down with 
a slight stippling movement, at the same time blowing gently 
into the rubber tube. Liquid will come out more easily if the 
tip is brought into contact with one of the larger drops of 
condensed moisture. This is usually unnecessary, unless the 
opening in the tip has a margin so even that it becomes closed on 
contact with the cover glass. 

If no particular individual bacterium is wanted, a simple way 
to isolate one organism is to take up some dozens of them, eject 
them on the cover, and add broth from the pipette. Then fill 
the pipette with this dilution and make a series of fine drops, 
in one or several of which a single cell will appear. Where the 
original mixture is not too thick, it is often easy to take any 
isolated organism alone into a fine pipette. If several enter, it 
is a small disadvantage, since they may be separated immediately. 

The droplet is made small so that one can easily assure himself 
that it contains but one organism. There is no danger of error 
if the droplet measures 25 microns or less in diameter. In such 
a droplet, for example, in peptone solution or any clear fiuid, 
one can make sure of the presence of but one actively motile 
cholera vibrio. There is no need for India ink or any addition 
to the fluid to make the bacterium more conspicuous. With many 
organisms, much larger droplets will meet the requirements 
(Plate I, figs. 1 and 2). If necessary, one may introduce the 
pipette cautiously to the edge of the droplet and remove the 
liquid, leaving the organism against the cover where it becomes 
more conspicuous. The isolation may be done with the low power 
and the droplets examined with the higher powers afterwards, 
or the whole process may be carried out under a high power, the 
oil-immersion lens if desired. For the isolation of bacteria of 
erdinary size, a 4-inch objective will suffice. 

If drops of moisture so large as to interfere with the making 
of the fine droplets have collected on the cover glass, a suitable 
area with fine droplets can almost always be found in the neigh- 
borhood of a larger drop of agar or broth. If a particle of 
doubtful nature is seen with the isolated bacterium in the droplet, 
it is usually easy to pick up either it or the bacterium and place 
each in a separate droplet. One may easily pick up the isolated 
bacterium and transport it to another part of the cover glass. 

The isolation of bacteria can be done very quickly by one 
experienced in the technique. I have performed the whole pro- 


322 The Philippine Journal of Science 1914 


cess, including the drawing out of the capillary and the making 
of the point, the adjustment under the high power, and the isola- 
tion of a micrococcus, in less than three minutes. 

Recapitulation of the various steps in the isolation of bacterium 
by method I: 

1. Clamp the pipette holder in position on the microscope. 

2. Prepare the moist chamber. 

3. Prepare and sterilize the cover glass. 

4, Seal the cover glass to the moist chamber and mark its 
upper surface. 

5. Place under the cover glass hanging drops of sterile nutrient 
fluid, and supply one of them with the bacteria to be isolated. 

6. Place the moist chamber on the stage of the microscope, and 
focus on its free edge with the low power. 

7. Make the capillary pipette. 

8. Attach the rubber tube to the pipette, and adjust the pi- 
pette in the holder with its tip in focus under the low power. 

9. Supply the pipette with sterile nutrient fluid from a hang- 
ing drop, and adjust it in the center of the high-power field. 

10. Take up the bacteria and isolate them in separate droplets. 


METHOD II 


Some investigators may wish to obtain one-cell cultures of > 
bacteria, fungi, alge, or microscopical animals without having 
occasion to do more extended selections. Such persons may 
hesitate to go to the delay and expense of obtaining a pipette 
holder. The following method will enable the worker to obtain 
pure cultures with the assistance of only such apparatus and 
materials as are found in every laboratory. The technique is 
but slightly more difficult, and very precise results may be 
obtained. The preparation and setting up of the apparatus 
needed may be done in one or two hours, and the technique may 
be mastered by an ordinarily skillful laboratory worker in half 
a day or less. 

The moist chamber, cover glasses, and tubing are the same 
as those used in the first method. In place of the pipette 
holder, an ordinary dissection microscope, supplied with rachet 
and pinion, is firmly clamped to the table as far from the edge 
as the clamp will allow (fig. 7). The arm of the lens holder is 
turned backward, and on it a rectangular perforated cork is 
firmly fastened with a clamp or rubber bands. The opening in 
the cork should be large enough to hold the glass tubing snugly, 
but not so firmly as to prevent the slipping of the tube back and 
forth. The capillary and tip are made as in the first method, 


IX, B, 4 Barber: The Pipette Method 323 


and the blunt end of the tube is slipped through the cork. If the 
tube is too small to fit well, a piece of paper may be rolled around 
it. The rubber tube is now adjusted and passed under a clip of 
the stage of the dissecting microscope, so that any slight move- 
ment of the tube will not disturb the pipette. 

The low power of the microscope is now focused on the free 
edge of the cover, then lowered somewhat as in the first method. 


til 


aa 


(FF 
—| 
AS 


x. 
Se 
| 
(ms) Ga) B 
e414 Fes 
es 
SA \n wea 54 
IN 


Fic. 7. A dissecting microscope used as a pipette holder. 


The compound microscope is slipped along the table until the 
center of the free edge of the moist chamber is near the tip of 
the pipette; then the pipette is raised or lowered until it is about 
midway in the center of the opening. By sighting along the 
edge of the cover and adjusting the movable arm of the simple 
microscope, one can bring the tip of the pipette into such a posi- 
tion that it can be found with the low power. In case it is not 
1284432 


324 The Philippine Journal of Science 1914 


readily found in this way, the pipette may be slipped in and out 
or moved back and forth until the tip comes into view. It is 
important to have the tip at a safe distance from the cover, 
because it is more liable to be broken with these less exact adjust- 
ments than is the case when the pipette holder is used. 

The tip may now be brought to the center of the field or to a 
point which corresponds with the center of the high-power field. 
The finer in-and-out adjustment may be made by slipping the 
tube with the fingers held in contact with the glass and with the 
top of the pillar of the dissecting microscope. The fine adjust- 
ment to and from the observer may be done accurately by sus- 
pending a pencil pendulum-wise from the fingers and Very gently 
tapping the end of the arm of the dissecting microscope on the 
one side or the other, as the case may require. 

The high power is adjusted as in the first method, and further 
adjustments to the center of the field are made by tapping the 
arm of the holder for one direction and moving the nose piece of 
the microscope for the other. Considerable right-and-left adjust- 
ment may be obtained by the nose piece alone without materially 
interfering with the illumination or definition. In some micro- 
scopes it may be necessary to loosen, temporarily, the clip fixing 
the position of the nose piece. 

The better the focusing apparatus of the simple microscope, 
the better the results obtained.. I have found no difficulty in 
isolating very small motile bacteria under the 4, oil-immersion 
lens by this method, and persons who have had no previous ex- 
perience in the isolation of microédrganisms by any method have 
succeeded in isolating blood corpuscles or bacteria after less than 
an hour’s practice. The adjustments once made, the technique 
is nearly as easy as with the holder; the method may be used 
with one type of compound microscope as well as with another. 
But the greater amount of time and care required for adjusting 
new pipettes make this method less suitable than the first for one 
who is doing an extended series of selections or inoculations. 
Further, the second method is less suited than the first for 
dissection under the microscope, inoculation into cells, or for 
some other special applications of the technique to be mentioned 
later. 


METHOD III 


In the third method of isolation, all mechanical holders are 
dispensed with and the pipette is held in the fingers of the 
right hand. The moist chamber is supplied with drops of 
sterile broth and of the bacterial mixture, as in the other 


IX, B, 4 Barber: The Pipette Method 325 


methods, and placed on the stage of the microscope with the 
open end to the right. The pipette is made as usual, though 
it is advantageous to have the turned portion rather short and 
brought to an angle of about 50 degrees instead of at right 
angles. If the low power alone is to be used, it is focused 
on the edge of a drop of sterile broth and then lowered some- 
what. If the high power is to be used, there is no preliminary 
focusing with the low power, but the lens is focused on the 
cover glass in the neighborhood of the edge of a sterile drop. 

The pipette is held with the thumb and index finger of the 
right hand and steadied by pressing the other fingers against 
and beneath the stage of the microscope. The shank is held 
horizontally and the tip brought into the chamber past the 
objective. It is then held far enough from the cover glass to 
avoid any drop and moved back and forth until the capillary 
can be seen, often only as a shadow in the field. The pipette 
is then slowly withdrawn until the tip appears. This is then 
kept in view while the objective is focused on the edge of the 
drop of broth. After the tip is filled from the drop, the chamber 
is moved and the bacteria brought into the field. The pipette 
may be filled from a test tube before introducing it into the 
isolating chamber. If it is necessary to break off the point, 
scratch it very gently on the side of the tube. The rapidity 
with which the broth rises is a good index of the size of the 
opening. 

The screws governing the mechanical stage and the fine adjust- 
ment of the microscope are manipulated by the thumb and second 
finger of the left hand, while the index finger is bent and braced 
against the pillar. The isolation is carried out as in the other 
methods. It is necessary to keep the point of the pipette con- 
tinually in view during the process. 

The advantages of this method are its simplicity and the 
rapidity with which the pipette may be adjusted into place 
and manipulated. The chief disadvantage is, obviously, the 
difficulty of properly governing the movements of the pipette 
with the fingers alone, especially under the higher powers. It 
is surprising, however, how steady the pipette can be held 
and how accurately it can be moved when the hand is well 
supported against the stage of the microscope. The isolation 
of organisms distinguishable under the low power is compara- 
tively easy, and the difficulty of isolating cocci and other smaller 
bacteria under a }- or 4-inch objective is not great after some 
practice. This method is not adapted to microscopes in which 
the fine adjustment and the screws of the mechanical stage 


326 The Philippine Journal of Science 1914 


cannot be governed by the left hand in the position described 
above. Its scope is also limited to the simpler applications of 
the pipette method. 


CULTIVATION OF THE ISOLATED ORGANISM 


After the single organism has been isolated, it may be 
cultivated in situ on the cover glass or removed and grown 
elsewhere. 


I. CULTIVATION IN SITU ON THE COVER GLASS 


1. If only a few generations of growth are to be observed, 
one may simply leave the organism in the droplet in which 
it was isolated and place the cover glass over an ordinary 
moist chamber. Some organisms do not grow well in so small 
an amount of medium, while others will form a considerable 
number of generations. Bacterium coli commune, for example, 
will form 32 small elements in a droplet of broth about 10 
microns in diameter. In order to prevent drying or undue 
concentration of the medium, the droplet should be placed near 
a larger drop of broth or of agar. It may be necessary to 
add to the moisture on the cover by placing slightly warmed 
water in the bottom of the isolating chamber just before trans- 
ferring the cover, but it is obviously unsafe to add too much 
condensed moisture in this way, because the drops may run 
together. 

2. With a fresh, sterile pipette (it is not necessary to make 
a fine-pointed pipette), liquid may be taken from a test tube 
or from a sterile hanging drop on the cover and added to 
the droplet containing the organism. This may be done under 
the low power. To avoid any possibility of taking up the 
organism with the second pipette, one may discharge the large 
drop near the small one and lower the pipette before the broth 
has spread to the droplet. Liquified gelatin, liquified agar, or 
any fluid or semifluid medium may be added. 

3. A very convenient method, especially when a considerable 
number of isolations are to be made from the same source, 
is to place on the cover glass previous to isolation a series 
of drops of broth, melted agar, gelatin, or any other solid or 
fluid medium. The diameter of these drops will depend on the 
available space and the nature of the experiment—0.5 milli- 
meter is, perhaps, the minimum and 2 or 3 millimeters a good 
average. These drops may be placed in a series of rows ar- 
ranged with reference to lines on the cover, or any arrange- 
ment may be followed, so that any drop can be easily found 


a oS 


IX, B, 4 Barber: The Pipette Method 327 


or registered according to its position (fig. 8). As many as 
from 50 to 60 drops may be placed on the same large cover. 
Each organism as it is isolated is placed in a small droplet 
close to any one of these larger drops ¢ (fig. 8), using the same 
pipette for all. When the series is finished and the small drops 
are examined to make sure that each contains but one organism, 
each small drop of a pair is made to fuse with the larger one. 
This may be done in several ways: (1) By means of a fresh 
pipette filled with sterile broth, fluid is added to the larger 
drop and the pipette withdrawn before the liquid spreads to 
the smaller. Or the smaller drop may be enlarged to meet 


the larger and the organism washed in. One may fuse all the 
pairs of the series with the same pipette. (2) If agar or a 
similar solid medium is used, one may pierce the large drop 
with a coarse pipette and, by moving the mechanical stage, 
slip the mass slightly until it meets the organism. This may 
also be done with a bent plati- 
num wire without the aid of a giuad 
the microscope. (3) When the | plese one | 
small droplet is very close to 0.185900 00 55) 000 00 00 oO} f) 
the large one, one may often 
fuse the two by adding water 
to the bottom of the isolating 
chamber sufficiently warm to fic.8. A cover glass with droplets arranged 
cause enough condensed mois- fo" an extended series of nations, 
ture to j oin the dr ops. bacterium-containing droplet beside it. 
The organism may be placed directly on the surface of the 
agar or gelatin, but the outlines of the droplet are less distinct 
and it is then more difficult to make sure that the droplet contains 
but a single organism. A drop of agar or gelatin may be placed 
near the center of a large cover, the organism placed on it, 
and a small cover pressed down on the organism. One may 
then observe the growth taking place between the two covers. 
The moist chamber to which the cover is transferred should 
be as shallow as the size of the droplets will permit, so that 
there will be little air for the absorption of moisture from the 
cover. A form convenient for large cover glasses may be made 
by cementing strips of glass to a large slide. Immediately 
before transferring to the cover, it is well to increase the mois- 
ture on the cover slightly by placing warm water in the bottom 
of the isolating chamber, and it is advisable to add condensed 
moisture in the culture chamber by breathing on the bottom 
of it. Unless the number of inoculated drops on the cover is 
large, there should be extra drops of broth placed on it to 


328 The Philippine Journal of Science 1914 


add to the moisture. After the cover is well sealed on, it is best 
to put the culture into a Petri dish or box, because a sudden 
change to a higher temperature may cause too much moisture 
to leave the cover and condense on the slide. These and similar 
precautions will occur to any one familiar with cultivation of 
organisms in the hanging drop; but special precautions must be 
taken where the drops are relatively small. 

The growth of the single organism at the edge or center of 
the hanging drop, whether in a liquid cr a solid medium, gives 
one an excellent opportunity of observing the development of 
young colonies. For instance, a single tubercle bacillus of some 
strains will give a very different type of growth in the bottom 
of a drop of broth than when left at the margin. If one 
desires to transfer the colony grown on the cover to a test tube, 
the cover is placed on the isolating chamber again and the 
colony transferred by means of the pipette under the low power 
or with a bent platinum wire without the use of the microscope. 

CULTIVATION IN A MEDIUM APART FROM THE COVER GLASS 

1. The organism may be taken up with a fresh sterile 
pipette already supplied with broth and discharged into a test 
tube containing a liquid medium, or it may be placed on the 
surface of a solid medium in a test tube or into the water of 
condensation. If it is desired to get an abundant growth on 
solid media as soon as possible, one may wash the surface with 
the water of condensation some hours after the organism has 
been transferred to it. The removal of the organism from the 
cover may be facilitated by adding a little sterile broth from the 
second pipette to the droplet just before taking up the organism. 
It is easy, of course, to transfer to a solid medium in a liquid 
state in order to obtain anaérobic conditions. 

2. Where temporary growth is to be observed, one may take 
up the organism in a fresh pipette and leave the pipette in 
the holder. After any desired interval of time, the isolated or- 
ganism with its offspring may be discharged on the cover and, 
after inspection, drawn into the pipette again. This method is 
sometimes convenient for organisms which do not grow well 
in a hanging drop. 

38. If it is desired to observe growth on the cover glass and 
the cover on which the organism is isolated is for any reason 
unsuitable, the organism may be taken into a pipette, the pipette 
lowered, and a new sterile cover, supplied with broth or agar 
droplets, substituted for the old one. One has only to raise the 
pipette and discharge the organism into any desired place on the 
new cover. 


IX, B, 4 Barber: The Pipette Method 829 


4, In some cases it is convenient to transfer a series of 
isolated organisms, each to a separate test tube, without the 
loss of time necessary for a change of pipettes at each transfer. 
For this purpose the large cover is slipped to the left, so that 
about 2 centimeters of the top of the isolating chamber is free. 
An oblong piece of mica, about 2.5 centimeters by 4.5 centi- 
meters in size and provided with a circular perforation about 
5 millimeters in diameter, is sterilized in the flame and placed 
over the opening with its free edge in contact with that of the 
cover. A small sterile cover glass is placed over this opening. 
The isolated organism is taken up in a pipette and deposited 
on the smaller cover. The cover is then taken up in sterile for- 
ceps and placed into a liquid medium or on the surface of a solid 
medium in a test tube or Petri dish. Ora flattened platinum loop 
is moistened in a sterile fluid and placed with the flat side in 
contact with the top of the small cover. The cover will adhere to 
it, and may be easily lifted and transported. A new small cover 
is now put over the opening and a second organism placed 
under it. If necessary, the mica may be sterilized in the flame 
before receiving another cover. With organisms very sensitive 
to drying, it is best to place a small drop of broth on the 
underside of the cover before placing it on the mica. Where 
such a drop is used, the mica may be placed on the left end of the 
box, as shown in fig. 9. The isolated organism on the larger 
cover will, in that case, be better protected against drying, 
and it will not be necessary to cover the opening on removal! of 
the mica for sterilization. It may be unnecessary to isolate 
organisms previous to transfer if the drop containing the 
bacteria is shallow and the bacteria relatively few, so that one 
can be sure that only one bacterium enters the pipette. Or, if 
a small drop contains a known number of bacteria, they may 
be removed and transferred one at a time. In any case, the 
bacterium may be inspected in a droplet on the small cover 
to make sure of its isolation. When these small covers are 
removed to a solid medium, opportunity is afforded for observing 
the growth of the bacterium between the cover and the medium. 

5. By attaching two holders to the stage of the microscope, 
two pipettes may be used simultaneously (fig. 15). An organism 
may be isolated with one pipette and immediately picked up by 
the other and transferred, or may be allowed to grow in the 
second pipette. By the use of either this method or the one 
described under 4, one may remove bacteria from a test tube, 
isolate a single organism, and place the isolated organism into 
a new test tube after a stay of only a minute or so on the cover 


330 The Philippine Journal of Science 1914 


glass. I have used these methods in the study of the growth 
rate of Bacterium coli commune.’ 
6. The organism may be drawn into a short capillary tube 


ah 
on ian 


Fic. 9. Two-movement pipette holder, pipette and isolation chamber arranged to illustrate 
method of transferring single isolated bacteria by means of a perforated mica plate. 
ms, mica plate; cv, small cover glass over perforation; z, large cover glass; p, pipette; 
g, up-and-down adjustment of holder moved by screw f; 7, to-and-fro movement regulated 
by screw s. (From The Journal of Infectious Diseases.) 


of very small diameter and the tube placed on the underside of 
the cover where growth may be observed under high powers—the 
oil immersion, if desired. Glass tubing is drawn out into a very 


* Journ. Infect. Dis. (1908), 5, 379. 


IX, B, 4 Barber: The Pipette Method SOL 


fine capillary, and this is cut into any desired lengths in a sterile 
Petri dish—1 centimeter is a convenient length. An ordinary 
pipette is made, moistened in sterile fluid, and the capillary made 
to cling to it as shown in fig. 10. Care must be taken not to get 
liquid into the short capillary. The pipette is adjusted in the 
usual manner. The capillary is partially filled by touching a 
small hanging drop of broth or other fluid medium, and then the 
organism is drawn in by capillarity. The pipette is then turned 
and the capillary deposited on the underside of the cover in a 
dry area or in a drop of fluid, as desired. I have grown yeast 
cells in a capillary so narrow that multiplication was possible 
only in the form of achain. Plague bacilli in the center of such 
capillaries have shown involution forms not observed in the 
control outside. 

Again, an ordinary pipette may be drawn to a very fine capil- 
lary, so fine that human red blood corpuscles must bend at the 
edges in order to lie in it. Organisms may be taken up in 
this capillary and the capillary 
flattened against the cover and 
observed under the oil immer- 
sion while still attached to the 
pipette. It is then possible to 
discharge all or a part of the 
contents of the pipette into a ™« a3 ek i Gn want a as 
hanging drop. The tip may be 
sealed by touching it to a hanging drop of sterile vaseline under 
the cover, then removed from the holder, and placed in the in- 
cubator. It may later be brought under observation again. 
The sealed tip may be broken off, under microscopical control, 
by means of a separate, coarse-pointed pipette held in a second 
holder clamped on the microscope. The organism may then be 
discharged into a hanging drop. 


INOCULATION INTO ANIMALS 


The isolated organism, or any small number desired, may be 
taken up and immediately inoculated into an animal, subcu- 
taneously, intravenously, or intraperitoneally. A pipette of 
slightly different construction is used for this purpose. Tub- 
ing of tough glass, not too thin walled, is drawn out into a capil- 
lary slightly thicker than that of an ordinary pipette. The tip 
is made and bent as usual, although the bent portion should 
have as a base some of the thicker part of the capillary. The 
pipette is supplied with salt solution or broth from a test tube. 
Care should be taken to avoid breaking off too much of the tip 


332 The Philippine Journal of Science 1914 


against the walls of the tube. It should be filled 4 or 5 mil- 
limeters back of the bend. It is then adjusted as usual and the 
organism taken from the droplet in which it is isolated. The 
organism must actually enter the tube and not adhere to the 
margin of the opening. The tip is now brought into a hang- 
ing drop of sterile broth or salt solution for a few seconds, so 
as to wash the organism well back from the tip. The pipette 
is removed from the holder, and is ready for inoculation. The 
index finger and thumb hold the capillary; the middle finger 
is extended and the end of it pressed against the bend of the 
pipette, in order to push the point through a fold in the skin. 
By blowing into the rubber tube, the liquid in the tube is in- 
jected, the liquid back of the bend serving to wash out the or- 
ganism. The point usually enters easily into the skin of ordinary 
laboratory animals, and leaves a wound scarcely discernible with 
a lens. If the opening in the pipette is too fine, there is some- 
times difficulty in forcing out the dose. In this case, one may 
enlarge the opening by breaking off a small portion against 
any sterile surface. It is obvious that a tip broken off obliquely 
will penetrate the skin much more easily than a blunt one. If 
desired, one may wash back the organism with sterile fluid in 
the test tube just before breaking off the tip; but, in any case, 
it is best to wash it a short distance back before removing it from 
the isolating chamber. Large animals with very thick skin may 
be inoculated in the mucous membrane of the mouth. I have in- 
oculated carabaos in this way. Any desired number of organisms 
may be counted out and inoculated in one dose; or one organism 
may be isolated, allowed to grow in the hanging drop, and its 
offspring inoculated, leaving one or more to grow in the hanging 
drop as a control. 

Special experiments,® in which the organism has been dis- 
charged into a nutrient medium instead of into an animal, have 
shown that the organism comes out of the pipette and does not 
adhere to the glass. Further, I have obtained a fatal infection 
in mice following the inoculation of a single anthrax bacillus *° 
and a considerable proportion of positive results with single 
plague organisms inoculated into monkeys and guinea pigs.1* 

A modification of the inoculation pipette is of advantage in 
some cases, for example, in the piercing of a very tough skin or 
in intravenous or intraperitoneal inoculation. Here the tip of a 


° Journ. Infect. Dis. (1909), 6, 634. 
© Toid. 
“This Journal, Sec. B (1912), 7, 251. 


IX, B, 4 Barber: The Pipette Method 333 


rather thick capillary is drawn out into a hair point, d (fig. 6), 
and the end cut off, as described in the technique of making ordi- 
nary pipettes. Only the hair point is turned up. This can 
usually be done by simply holding it above the flame of the 
microburner. The upward draft of hot air softens the glass 
and bends it vertically. This pipette is supplied with liquid; 
the organism is taken up and washed well back as described for 
the ordinary inoculation pipette. Then the hair point is broken 
off, and one has a straight, very sharp-pointed needle with the 
dose back of the tip and enough liquid present to wash it out. 
Such pipettes with the contained dose may be sealed at the tip 
and kept until a convenient time for inoculation. This modifi- 
cation has been used in the inoculation of doses of Bacillus tuber- 
culosis consisting of one or few bacilli? If desired, the dose 
may be discharged into the needle of a syringe and inoculated 
in that way. The pipette is gradually withdrawn from the 
needle as the dose is discharged into it. Semifluid agar or other 
substance may be placed in the syringe and in the base of the 
needle in order to carry out the dose. In most cases it is unnec- 
essary to use the syringe. The technique of inoculation into 
living cells will be described below. 


DETAILED DESCRIPTION OF CERTAIN STEPS IN THE ISOLATION 
METHODS 


MOISTURE 


The necessity of keeping the proper amount of moisture in the 
isolating chamber cannot be overemphasized. Drying or over 
concentration of the medium is fatal to some organisms, besides 
increasing the difficulty of isolation. The index of the proper 
amount of moisture is the presence on the cover of very small 
droplets of condensed moisture, the film resembling fine stip- 
pling (photomicrograph, Plate I, fig. 1). This stippling of con- 
densed moisture also aids the eye materially in locating the 
edge of droplets. Much depends on a proper amount of vaseline 
on the cover (see page 314). If large droplets of condensed 
moisture are formed, it becomes difficult to make the small 
droplets necessary for isolation. In this case one usually finds 
a suitable area in the neighborhood of a large hanging drop. 
One can usually supply the necessary film of moisture by placing 
slightly warmed water in the bottom of the isolating chamber. 
Distilled water may be kept conveniently at hand in a small 
reagent bottle closed with an ordinary medicine dropper. The 


% Journ. Med. Research (1909), 20, 1. 


334 The Philippine Journal of Science 1914 


dropper is filled and the water in it heated slightly over the 
flame. The temperature of the water is gauged by the tem- 
perature of the room. One can usually judge the condition of 
the moisture film with the naked eye. As a rule, it is best not 
to use the outer third of the cover for isolation. By keeping this 
outer third of the cover supplied with hanging drops of agar 
or broth, the moisture of the other two-thirds of the box may 
be increased. Very constant conditions of moisture may be 
obtained within a long, narrow area on the cover inclosed by a 
barrier of agar. It is important to have the cover well sealed 
to the edges of the chamber in order to avoid convection currents. 
Drafts of air on the laboratory table should be avoided, especially 
those blowing toward the open end of the chamber. 

One may protect the open end of the box by a piece of 
moistened filter paper provided with a slit for the capillary por- 
tion of the pipette. A hood of moistened blotting paper (H, fig. 
1) is still more convenient. A slight modification of the isolating 
chamber will assist in protecting the under surface of the cover 
in special experiments. A glass strip is cemented to the slide 
just outside of the chamber in such a way as to form a groove 
into which a piece of moist blotting paper may be inserted in 
an upright position, or the blotting paper may be slipped into 
the chamber and held upright by the sides. This method of 
protection leaves less working room for the pipette, but this 
difficulty may be avoided by bending the end of the pipette into 

the form shown in fig.11. Here 

the tip of the pipette is just 

above the line of the capillary, 

and this arrangement allows one 

to work with the capillary of the 
Fig. 11. A pipette constructed for use above pipette above the barrier of 

a barrier of blotting paper. 5 

blotting paper. 

I have used an isolating chamber, a portion of which is wholly 
sealed by means of a trap filled with water or mercury, t (fig. 
12). The pipette capillary is bent into the form illustrated in 
the figure and adjusted into the chamber. If a capillary of hard 


Fic. 12. Special isolating chamber in longitudinal section. The chamber b is closed at the side 
by the trap ¢ and partition p. 


IX, B, 4 Barber: The Pipette Method 3835 


glass is used, little difficulty will be found in bending it into any 
desired form over a small flame. Then the partition p, which 
fits into grooves at the side of the isolating chamber, is slipped 
down into place, and the cover is put into place and sealed with 
vaseline. An oxygen-absorbing reagent may be placed in the 
compartment 6 and a wholly or partially anaérobic condition 
attained. 

Such a device is unnecessary for ordinary work, and even the 
moist paper protecting the open end can usually be dispensed 
with. If the bottom of the chamber is well supplied with water 
and the cover well sealed on, small droplets will remain on the 
inner half of the chamber for hours, even when the end is 
uncovered and the apparatus unprotected by a bell jar. By 
taking very simple precautions, one can work in a very warm 
and dry room. 

DANGER OF CONTAMINATION 


Danger of contamination is very small, less than when ordi- 
nary plate cultures are used, as there is no strong current of 
air entering the chamber. Droplets of broth remain sterile for 
many hours with the end of the chamber open and unprotected. 
In work where special precautions are to be taken, one may 
protect the end of the chamber with moist filter paper or the 
paper hood while the pipette is in place, and at other times keep 
the chamber on a glass plate protected by a bell jar or crystal- 
lizing dish. 

PREPARATION OF ORGANISMS FOR ISOLATION 


As stated above, no preliminary dilution of the organisms to 
be isolated is necessary. The material may be taken from any 
source, and the emulsion of organisms may be of any density. 
The organisms to be isolated may be in the majority, or they may 
be surrounded by thousands of other organisms of the same or 
another species. A single red corpuscle may be easily isolated 
from undiluted blood. In selection experiments, one may sep- 
arate any aberrant organism from myriads of others of the same 
sort. The only requirement is that the organism is recognizable 
and large enough to be seen under an oil-immersion lens. Ani- 
mals, unicellular alge, or spores of fungi may be removed from 
a thick emulsion of bacteria, washed with the pipette, and isolated. 
Bacteria may be separated from pus or other pathological 
material. 

There is always the possibility that a nonmotile organism 
may not be viable. One can often save the labor of making 
a long series in search of a living organism by a short prelim- 


336 The Philippine Journal of Science 1914 


inary cultivation. The material is placed in a hanging drop, and 
some suitable nutrient material is added. The drop is inspected 
and set aside for a preliminary growth. Living organisms may 
then often be recognized by the chains or groups formed in 
multiplication. By isolating such a group and selecting an in- 
dividual from it, one is practically sure of obtaining a viable 
organism. Spores of fungi may be left until the hypha of 
germination has begun to form, and then they may be isolated. 
This preliminary growth need not be continued for more than 
an hour or two in the case of many bacteria and fungi. One 
may make a preliminary separation of motile organisms by 
placing the material containing them at one end of a long hang- 
ing drop. They may be picked up after they have swarmed to 
the farther side of the drop. This same device may be used 
in the partial separation of animals or zodspores of fungi or 
of alge from bacteria. 

A more motile organism may be partially or wholly separated 
from a less motile one by this method. By using a peptone solu- 
tion, cholera vibrios in a drop of feces may often be separated 
sufficiently for an agglutination test. 

The separation of spores, cysts, or living animals from bac- 
teria is usually easy. The larger organism is isolated and placed 
in a droplet, the smaller the better. The bacteria in the end of 
the pipette are now discharged on the cover at a convenient 
distance from the hanging drop, and sterile fluid is added to the 
isolated organism. If desired, this fluid may be taken from a 
large hanging drop on the cover. The liquid surrounding the 
spore or animal is now removed and fresh liquid is supplied, 
or the organism may be picked out and discharged in a new 
droplet. This process is repeated until the organism is washed . 
free from bacteria. The process is usualy done most conven- 
iently under the low power. The organism in a small hanging 
drop may be inspected with the high power before being trans- 
ferred. If necessary, a few remaining bacteria may be removed 
from the spore with a finer pipette, even loosening them from 
the side of the larger organism. If the bacteria are embedded 
in some gelatinous material surrounding the spore or animal, 
it is sometimes impossible to remove them, but I have had no 
difficulty in wholly freeing amcebe, in cyst or motile condition, 
from bacteria. The process of washing the spore or animal 
usually requires only a very few minutes. 

A bacterium may be washed free from any animal exudate in 
this way. I have obtained successful infections with doses of 
a single plague bacillus and of a single anthrax bacillus which 


TX B; 4 Barber: The Pipette Method 337 


were washed in salt solution before inoculation. In washing 
small organisms, a fine pipette is brought to the edge of the drop- 
let containing the cell and the liquid is removed, leaving the cell 
in contact with the cover. Fresh liquid is substituted, and the 
process repeated as often as desired. 

It is often difficult to remove bacilli of tuberculosis and some 
bacterial spores from a hanging drop after they have settled to 
the layer of surface tension at the bottom of the droplet. The 
liquid passes into the pipette, but the cell remains stranded on 
the cover. This difficulty may be avoided by taking up these 
organisms before they have settled, or by placing the mouth of 
the pipette directly over the cell. SBacilli of tuberculosis may be 
made to cling to the side of the tip of the pipette and transported 
in that way. The peculiar behavior of these organisms may be 
due to the fat or mucus surrounding them, as most cells do 
not give this difficulty. It is difficult to remove leucocytes from 
a hanging drop of blood or any dilution rich in serum when they 
have once settled to the bottom of the hanging drop. They may 
often be isolated by bringing the mouth of the pipette directly 


under them. 
NUTRIENT MEDIA 


All media of the ordinary sorts may be used in this method, 
and where only small quantities are needed one has a wide range 
of possibilities. Quantities of medium may be repeatedly taken 
from the same test tube if one takes proper precautions against 
contamination. A soft agar can be conveniently obtained by 
melting the very top of a slant and allowing a portion of the 
liquified agar to flow into the water of condensation. By mixing 
different proportions of the liquified agar with the water of con- 
densation, one may obtain various degrees of stiffness. Suffi- 
cient liquid for small hanging drops may be taken from a small 
blister or from the body of an insect or other animal; or, where 
the inoculation pipettes are used, from the contents of a living 
plant cell or microscopical animal. If a small quantity of serum 
free from corpuscles is needed, a deep hanging drop is made of 
the whole blood, and, when the corpuscles have settled to the 
bottom of the drop, serum may be drawn from its margin. A 
bacterium-free liquid from the cultures of some microdrganisms 
may be obtained in the same way. 


ILLUMINATION 


The illumination of objects on the cover during isolation may 
be improved by the use of a projection condenser which focuses 
the light some distance above the top of the stage. Hither day- 


308 The Philippine Journal of Science 1914 


light or artificial light may be used, but a fairly strong illumina- 
tion is advisable, especially when the higher powers are used. 
The oil immersion can be readily used over the isolating chamber 
without resorting to any special illumination. 


PIPETTES 


Pipettes may be made some time before use and kept in a 
sterile chamber or placed in a sterile test tube or flask with 
the shank held in place by the cotton plug, somewhat as shown 
in fig. 16. It is usually more convenient, however, to make the 
pipette just before use. A very convenient plan is to close the 
pieces of glass tubing intended for pipettes at one end with 
cotton and to draw them out into a coarse capillary at the 
other. The capillary is sealed at the tip, and the pipette steril- 
ized in the hot-air sterilizer. A number may be sterilized 
and kept in stock ready for use. The coarse capillary may be 
drawn out and made into a tip for isolating, or may be simply 
turned up in the Bunsen flame and broken off for use in placing 
the preliminary drops of sterile liquid or solid media on the 
cover. 

GAS FOR THE MICROBURNER 

When natural gas is used, it is sometimes difficult to keep 
the small flame lighted. This can be remedied by causing the 
gas to pass through a bottle containing benzine or other en- 
riching fluid. 


SPECIAL APPLICATIONS OF THE PIPETTE METHOD 


ISOLATION OF ORGANISMS FROM QUANTITIES OF WATER LARGER THAN 
HANGING DROPS 

Sometimes it is desirable to isolate alge, protozoa, bacteria, 
or other organisms from water or from a culture in which 
they occur in such small numbers that one cannot find them 
readily in the hanging drop. In this case, a Petri dish, a watch 
glass, or other receptacle containing the organisms may be placed 
on the stage of the microscope and a pipette, bent as shown 
in fig. 18, may be adjusted in the holder. The capillary need 
be bent only enough to avoid the edge of the dish. The point 
of the pipette is made as fine as desired, the opening being suited 
to the size of the microérganism. The tip is bent down obliquely, 
if the organisms sought are likely to be at the bottom; or bent 
upward, if they are to be sought at the surface. It is possible 
to scrape the bottom with the pipette and loosen and take up 
any adhering amoebe or spores of alge. When the organism 
sought is taken into the pipette, the tip is raised above the surface 
of the water, the Petri dish removed, and an ordinary isolating 


IX, B, 4 Barber: The Pipette Method 839 


chamber, supplied with a cover, is placed on the stage. The 
pipette is then turned point upward and adjusted so that it 
may enter the chamber. The tip is focused under the low power, 
and the organism is discharged into a hanging drop on the 
surface of the cover. It may then be washed free from bacteria 
or other organism if desired; it may be grown on the cover or 
transferred to a watch glass, a test tube, or other receptacle. 
The low powers of the microscope are most convenient for 
this work, but a + objective may be used for smaller organisms. 
In this case, a flat piece of cork is perforated with an opening 
about 1 centimeter in diameter and the opening is enlarged funnel 
‘wise on one side so as to admit the objective. A cover glass 
is sealed with vaseline to the other side of the cork, and the 
float is placed cover side down on the water. One may adjust 


Ve 


Fic. 13. A special pipette in position for isolating organisms from the bottom of a Petri dish. 
p and pa, metal plates screwed to the stage for the attachment of the pipette holder. The 
complete pipette holder is used, but only a portion of it is shown in the figure. 

a pipette under the float and focus the higher power of the 

microscope on the tip and the liquid surrounding it. If it is 

desired to select organisms deeper in the liquid with the high 
power, a short, thick rubber tube or hollowed rubber stopper 
is placed around the base of the lens and the free margin smeared 
with vaseline. A large cover is placed on a dry slide, and the 
objective with the rubber tube is lowered to the cover. The 
smeared edge reaches the cover and seals fast to it, and the 
lens is made to descend until one can focus below the cover. 

Any stained object on the slide will facilitate the focusing. On 

raising the objective, the cover comes up with it; then it may be 

lowered into the water containing the organisms to be isolated. 

If organisms cling to the cover on bringing out the objective, 

they may be picked off from the cover with the pipette, or 

the pipette may be adjusted and isolation carried out as in the 
case of the cork float. 
1284433 : 


340 The Philippine Journal of Science 1914 


FIXATION AND STAINING 


Fixation ald staining can be carried out in hanging drops 
with the pipette. The stain or fixing reagent may be placed 
under the cover with a platinum loop or coarse pipette and 
applied with the isolating pipette to the organism. The medium 
in which the cell lies may be withdrawn and the cell washed, 
following the method outlined for freeing cells from bacteria 
or serum (see page 837). The fixative, and then the stain, 
may be applied with the pipette. The same pipette may be 
used for both by washing it between each use with water or 
other fluid in hanging drops. If the fixative is volatile or can- 
not be easily washed out of the pipette, one may keep it in a 
separate pipette. A second holder may be used or the pipette 
removed from the holder and the one containing the stain 
substituted. After staining, the cell may be washed and de- 
colorized if necessary. If it is desired to mount the stained 
preparation in ‘balsam, albumen or other fixative may be ap- 
plied and the location of the cell marked by Brunswick black 
or India ink on the upper side of the cover. The preparation 
is then dehydrated by drying on alcohol and mounted. Or the 
cell or colony may be located, and the cover removed, dried, and 
stained. In Plate I, figs. 3-7, a photomicrograph is shown of 
a preparation of Bacteriwm coli. By means of successive iso- 
lations, a series of 5 droplets was obtained, containing respec- 
tively 1, 2, 4, 8, and 16 bacilli. The preparation was dried in 
toto, stained, washed, and mounted without the loss of a single 
bacillus in the series. I have also used this method in staining 
young colonies of Bacillus tuberculosis derived from an isolated 
single cell. Drops of different stains may be dried on a large 
cover. When they are to be used, the cover is placed over 
an isolating chamber, and the stain, dissolved in the water of 
condensation, is taken up with the pipette. By changing covers 
and using two pipettes, one may obtain a great variety of 
combinations. 

SEROLOGICAL TESTS 


I have tested only agglutination and precipitation by the pipette 
method. The agglutination in hanging drop is especially practi- 
cable with motile bacteria rapidly agglutinated, such as vibrios of 
Asiatic cholera and typhoid bacilli. The serum in proper dilution 
is taken into a pipette and applied to the edge of a hanging drop 
containing the bacteria. This method gives very striking results 
with vibrios. When a very small quantity of cholera agglutinat- 
ing serum is applied at the edge of an emulsion of cholera vibrios, 


eT 


a eee ee eee ee ee eee 


IX, B, 4 Barber: The Pipette Method 841 


a ring of agglutinated vibrios forms instantly and all vibrios 
which come into this ring are agglutinated at once. If some 
other vibrio is substituted for cholera, not only does agglutination 
with the cholera specific serum fail, but the vibrios are attracted 
by the serum. A good method is first to make a drop of diluted 
serum very near the bacterial emulsion, then to connect the two; 
the serum will flow through the connecting channel into the larger 
drop. 

This method has been found especially practical in testing out 
a large number of colonies on a plate. A series of drops of salt 
solution, peptone water, or other fluid is made on a large cover 
glass placed over the isolating chamber, and bacteria from various 
colonies are added to them. Different agglutinating serums may 
be placed in hanging drops on the same cover or on another 
cover placed over a second moist chamber. All drops may be 
tested with a given serum, the pipette washed in a hanging drop 
of salt solution, and a second serum applied if necessary. If 
desired, the serum may be used in low dilution in order to elimi- 
nate negative colonies. Those which give a positive or doubtful 


_ test may be further tested by the macroscopical agglutination 


method. One may dispense with the pipette and test by placing 
the serum on the under surface of the cover with the loop and 
connecting it with the emulsion drop to be tested, but the pipette 
gives more precise results, and is more convenient, especially 
where a large number of tests are to be made. 

By the same general method a precipitating serum may be 
added to a hanging drop of the dilution to be tested. The forma- 
tion of the precipitate may be observed microscopically. Appli- 
cation of this technique to other serological work as well as to 
various microchemical tests are possible, but I have had no ex- 
tended experience in any but the above. 


EXPERIMENTS ON CHEMIOTAXIS 


Experiments of this nature may be carried out with the tech- 
nique described under serological methods, or by using the 
capillary tubes in flasks as described on page 346. It would 
seem possible to test the effects of currents of electricity in 
hanging drops, with or without organisms. Two pipettes may 
be used for the two poles by filling them with mercury, as in the 
technique of inoculating cells. One could insulate one pipette 
by drawing some nonconducting liquid into the lumen and, by ap- 
plication of pressure to the pipette, eject the nonconductor and 
make a mercury connection. If desired, this process may be 
conducted with both pipettes in the same animal or living vege- 


342 The Philippine Journal of Science 1914 


table filament. These methods have not been tested by me, and 
are given only as suggestions. 
DILUTIONS 


It is possible to make dilutions of a serum or other fluid under 
the microscope with the pipette. A pipette with a tip bent some- 
what backward is made, and a mark is made on the capillary 
below the point of the tip. The pipette is filled to the mark with 
serum by dipping the tip in a hanging drop, and then the mark is 
brought into focus with the low power. The contents are ejected 
in a hanging drop, and an equal quantity of salt solution is added. 
These are mixed in the pipette, and one or more portions of the 
first dilution are added to one or more of salt solution, and so on. 

If desired, a hanging drop may be made to contain a measured 
quantity of liquid. For measuring, a special pipette is made (fig. 
14). This capillary is made of hard glass, preferably, and after a 
point is made and turned the capillary is bent into the position 
shown and the upper side of the triangle is marked with a very 

fine capillary or pen dipped into 

India ink or asphalt cement. 

After the pipette is placed in the 

holder, one can estimate the 
Fic. 14. Special pipette for measuring small cubic contents of the different 

quantities of liquid. a 

segments of the capillary by 
measuring the length and diameter of each with the eyepiece 
micrometer. The tip is brought into the center of the low power 
and filled from a large hanging drop. By focusing down and 
moving the nose piece, or the right-and-left adjustment of the 
pipette holder, to and fro, one can bring the different marks into 
view and discharge a hanging drop containing the quantity lying 
between any two marks. If desired, the contents of a given 
portion of the capillary may be more exactly measured by filling 
it with mercury and weighing the amount of mercury ejected 
when it is emptied. 

It is hardly necessary to state that these measuring methods 
are applicable only when the amount of condensed moisture on 
the cover is constant and small. However, the method has been 
found useful in making a series of drops of approximately the 
same size. If very small drops are needed, the eyepiece microm- 
eter may be focused on the capillary and used in determining 
the length of the column of liquid to be ejected. 


WARM BOX 


A series of isolations can be made with the microscope inclosed 
in a warm box. In an apparatus used by me for determining 


IX, B, 4 Barber: The Pipette Method 343 


the growth rate of bacteria, the microscope was inclosed in a 
wooden box which was covered with a cloth jacket lined with 
asbestos wool. It was warmed by electric coils, and the temper- 
ature was regulated by a thermoregulator. The adjustment 
screws of the pipette holder were reached through a small door 
in one side, the rubber tube was passed out through a small 
opening at the top, and the mechanical stage was moved by 
means of rods passing through openings in the top of the box 
and engaging the screws of the stage. With this apparatus it 
was possible to keep the moist chamber at a constant temperature 
throughout an experiment lasting a day or more. A bacterium 
was isolated, and when it had developed two or four offspring 
a daughter cell was isolated, placed in a new drop, and so on. 
Thus a record of the number-of generations formed in a given 
time was kept. The method in which the movements of the 
pipette are governed by a simple microscope or other holder 
’ apart from the compound microscope would be adapted to a warm 
box too small to admit both microscope and holder. 


DISSECTION 


Some of the simpler dissections in the hanging drop may be 
done with a single pipette. In order to break apart a clump 
of bacteria, one may draw away the liquid surrounding the 
clump and press apart the bacteria with the pipette tip. A 
yeast cell containing spores may be broken open and the spores 
separated with the ordinary blunt-pointed pipette. With the 
pipette drawn to a sharp point (fig. 6), other manipulations 
are possible. An amceba, when stretched out on agar, may be 
cut into two living parts, with little loss of cell contents, by a side 
stroke with the sharp point. This process is easier if the tip 
is bent somewhat obliquely as shown in ¢ (fig. 19). The nucleus 
of an amceba may be removed by introducing the point close 
to the nucleus when the latter is at the margin of the animal. 
A side movement, made with the mechanical stage, removes the 
nucleus with little loss of protoplasm, and the enucleated amceba 
continues its movements. The tip of the dissecting needle may 
be broken off and transformed into a pipette for staining or other 
purposes. 

I have succeeded in breaking open human red blood corpuscles 
with a single point, the size of which bore about the same relation 
to that of the corpuscle as the blunt end of a pencil does to 
the palm of the hand. 


* Journ. Infect. Dis. (1908), 5, 379. 


344 The Philippine Journal of Science 1914 


Two dissecting needles may be employed by the use of the 
two pipette holders. The second holder may be clamped to an 
elongated plate screwed to the stage of the microscope (fig. 
15) or to any firm support apart from the microscope. In 
order to bring both points into the moist chamber, a bend in one 
of the capillaries may be necessary. I have used an isolating 
chamber with a side opening, so that the second pipette holder 
could be clamped on the front of the stage. However, it was 


Fic. 15. Microscope with two pipette holders, each containing a pipette attached to the stage 
by means of metal plates. Seen from the back. A, a three movement, and B, a two move- 
ment holder; tf, adjustment, governed by screw s’’, for moving the pipette to and from the 
observer. The other letters correspond to those shown in Figs. 1 and 2. 


found difficult to keep the isolating chamber moist with two 
openings in it without using a special closing device, so the 
position first described is preferred. 

With both holders provided with three movements, one has at 
command two dissecting needles, with rigid points as fine as 
the end of a colon bacillus, each of them movable in all directions 
of space. In addition, it is possible to move the organism in 
two directions by means of the mechanical stage. Dissection 
may be carried on under the oil immersion, and either needle may 
be transformed into a pipette at will. The wall of Spirogyra 
may be opened and the nucleus removed, microscopical animals 


| 
. 


IX, B, 4 Barber: The Pipette Method 345 


dissected, and various other manipulations performed. Either 
one or both of the pipettes may be provided with mercury and 
used for cell inoculation. It is better to use hard glass in making 
fine points for dissection. 

For the dissection of very delicate, soft objects, as blood 
corpuscles and some protozoans, I have made use of sharp stiff 
hairs, taken from the body of a house fly, and also of very 
fine-pointed needle crystals. A pipette with a fairly large open- 
ing is made, and the hair or crystal drawn partially into it, the 
thicker end within. The fine point projecting from the tip of 
the pipette is then used as a probe or dissecting needle. The 
same pipette may be used as a holder for a considerable variety 
of these fine tools. 


THE PIPETTE AND ISOLATING CHAMBER USED AS SEPARATE UNITS 


There are some uses of the pipette or of the isolating chamber 
as separate units which may merit a short description. 

The isolating chamber has been found a very useful adjunct 
to the microscopist in furnishing a convenient method of examin- — 
ing material in hanging drop. One has only to place with a 
loop a drop of the material to be examined under the cover 
glass, and the same cover glass may be used for the examination 
of many colonies of bacteria, of samples of feeces, of protozoans, 
or of microscopical plants. If desired, the cover glass with 
the samples may be sealed on a ordinary moist chamber for 
further observation. A pipette containing an agglutinating 
serum or some simple stain may be used in connection with the 
hanging drops. 

The pipette will often be found convenient in fishing colonies 
from a plate in which the colonies are too close together to be 
transferred with the platinum needle. The Petri dish or other 
plate may be held in a clamp in an inverted position over the 
stage; where this is not practicable, an area of the medium 
containing the colonies may be transferred to a large cover and 
placed over an isolating chamber. 

A straight, fine-pointed pipette provided with a rubber tube 
may be used for inoculating microdrganisms into insects. .A 
point much finer than that of a syringe needle may be made 
and materials inoculated between the joints of the leg or into 
any part visible under a hand lens. If a very fine point is 
needed for inoculations into.animals or plants, the pressure 
apparatus described in connection with the cell injection tech- 
nique may be used. In order to gain freedom of movement, 
the pressure pipette may be suspended by a cord. The raising 


346 The Philippine Journal of Science 1914 


and lowering of the cup containing cold water may be done 
by an assistant. I have successfully inoculated defibrinated 
human blood into living mosquitoes by this method. 

A special application of the pipette may deserve a fuller de- 
scription. In this technique the separation of motile bacteria is 
accomplished by the use of pipettes supplied with fluids attrac- 
tive or repulsive to certain species of bacteria. For certain 
experiments specific agglutinat- 
ing serums may also be used. A 
short glass tube is plugged at 
one end with cotton and drawn 
out into a capillary at the other. 
It is then sterilized in the hot- 
air sterilizer. An ordinary pip- 
ette is made from it, supplied 
with the fluid to be used, and is 
placed in a test tube or flask con- 


as shown in fig. 16. A perfor- 
ated cork may be used in place 
of the cotton plug, and, in either 
case, two or more pipettes may 
be introduced into the same 
flask. The pipette is supplied 
with liquid from a test tube and 
more drawn in than can be sup- 
ported by capillarity. The liquid 
flows out of the capillary until 
only the column supported by 
capillarity remains. The slow 
diffusion of the liquid from the 
point forms a zone of attraction 
for motile bacteria positively 
chemotactic to the fluid in the 
pipette, and they may finally 
enter the pipette and grow in 
the capillary. It is easy to transfer them by removing the 
cotton plug from the pipette and withdrawing some of the liquid 
at the top of the column by means of a fine glass capillary or a 
long platinum needle. If it is desired to continue the use of the 
apparatus after the removal of liquid for examination, the pip- 
ette may be raised slightly after withdrawal of the liquid, so 
that no additional bacteria will be drawn into the tip by capil- 
larity, or new liquid may be introduced into the capillary. 


Fic. 16. Apparatus for attracting certain 
motile bacteria into pipettes. 


taining the mixture of bacteria, | 


IX, B, 4 Barber: The Pipette Method 847 


I have used this apparatus in some experiments with mixtures 
of cholera vibrios and Bacillus pyocyaneus. A pipette charged 
with a dilution of cholera agglutinating serum will attract only 
B. pyocyaneus, while a pipette supplied with B. pyocyaneus ag- 
glutinating serum will furnish pure cultures of cholera from the 
same mixture, even though B. pyocyaneus has so far over- 
grown the other that the cholera can no longer be detected by 
the plate method. If the bacteria are not in pure culture in 
the pipette, one or the other species may be so far in the majority 
that it is easy to separate them by the plate method. The experi- 
ments with cholera mixed with other bacteria are as yet un- 
finished, but the technique is far enough developed to justify a 
brief preliminary description here. There are many variations 
possible in this technique, some of which will be described in a 
subsequent article. 

The flask, shown in fig. 16, is a very convenient holder for 
pipettes temporarily removed from the pipette holder. The tip 
of the pipette is left just above the surface of the liquid, so that 
it will remain moist. Agglutinating serums, stains, or other 
fluids may be kept for days in this way, ready for use at any time. 


INOCULATION INTO LIVING CELLS “* 


The technique of inoculation of microdrganisms, stains, fix- 
atives, or other substances into the protoplasm or vacuoles of 
living cells is subject to some requirements not found in the 
simple isolation of cells. The tip of the pipette must be of such 
a fineness as to minimize the injury to the cells inoculated and 
of sufficient rigidity to pierce the cell wall. An injection force 
has to be employed sufficiently great to overcome cell pressure, 
capillarity in the very fine tip, and any obstruction in it. A 
pipette point, such as is shown in a, fig. 19, will meet the first 
requirements, and the necessary force for injection is obtained 
by the expansion of mercury. The isolating chamber, pipette 
holder, and microburner are the same as those described under 
method I. 

The special pipettes may be made of the same sort of tubing 
as for isolating pipettes, although glass with a slightly thicker 
wall (about 0.7 millimeter in thickness) is preferable. Hither 
hard or soft glass, if tough and of good quality, may be used. 
A piece of tubing about 35 centimeters in length is bent at one 
end into the form shown in fig. 17. The distance from the top 
to the bottom of the curved portion should be about 5 centimeters. 


* First described in Journ. Infect. Dis. (1911), 8, 348. 


348 The Philippine Journal of Science 1914 


loop 


Fic. 17. Pipette for injecting sub- 
stances into living cells. Com- 
pleted. 


For convenience in description, the 
curved portion will be designated as 
the loop and the straight portion as 
the arm. It is not essential to have 
the exact form of loop as that rep- 
resented in the figure. Sufficient 
bends should be made to contain a 
considerable quantity of mercury 
and to make the instrument sen- 
sitive to changes in temperature. 
Too many bends make it heavy and 
unwieldy. ‘ 

The tip n, drawn out into a coarse 
capillary, is inserted into a cup of 
mercury and the whole tube nearly 
filled with mercury by exhausting 
the air at the straight end. This 
may be done conveniently by suction 
through a rubber tube attached to 
this end. The mercury should be 
clean and dry; it is best drawn into 
the tube when hot. The tube should 
be heated before filling, and all pre- 
cautions should be taken to avoid 
the introduction of bubbles of air 
or of water vapor. After the tube 
is filled to a point about 2 centi- 
meters from the blunt end, the arm 
should be inclined so as to leave the 
capillary at n free from mercury. 
Now constrict the capillary in the 
flame at a point very close to the 
loop until only a very fine lumen 
remains. Raise the arm, and seal 
the constricted portion just as the 
outflowing mercury reaches it. By 
this method very little, if any, air 
remains at the tip. Loss of mer- 
cury from the open end of the arm 
may be avoided by temporarily plug- 
ging the opening with cotton. 

Draw out the end of the arm into 
a straight capillary about 8 centi- 
meters long and 0.5 to 0.8 millimeter 


IX, B, 4 Barber: The Pipette Method 849 


in outside diameter—the same dimensions as for an isolating 
pipette, although it is better to have the wall of the capillary 
slightly thicker (about 45 microns in thickness). 

The pipette should now be filled with mercury to a point ,well 
within the capillary. This is done by heating the arm at about 
point s (fig. 17) sufficiently to vaporize some of the mercury 
and expel all air from the capillary. Dip the capillary into 
mercury, and it will fill as the mercury in the arm condenses 
and cools. If air bubbles remain in the arm or loop, they may 
be worked out. Freeing from air is facilitated by heating the 
arm and sealing the tip while the mercury is vaporized at the 
point heated. On cooling, a vacuum will be left at the end 
and the air may be more easily worked out. When the capillary 
is opened again, it may be necessary to refill it with mercury. 
The pipette should be as free from air as possible, although 
a small bubble does not prevent its successful use. Time will 
be saved by making two or three pipettes at a time. One may 
be carried through a stage of the process while another is cooling. 
After they have been finished to the stage described, they may be 
set aside until a time convenient for use. 

When ready for the inoculation, the cells into which substances 
are to be injected should be placed in a shallow hanging drop 
near the center of a large cover which is sealed over the isolating 
chamber, as for ordinary isolation. Sufficient moisture must be 
supplied. If the cells or filaments are in pure culture, as a fish 
mold in agar or broth, it is well to have a round or oval paraffin 
barrier under the cover. This is traced on the cover by means 
of a coarse bent pipette containing melted paraffin. The fungus 
may be cultivated within this barrier a day or two before in- 
oculation. The barrier protects against contamination with 
bacteria.° Animals, alge, or fungi not in pure culture may be 
placed under the cover with no special precautions. A hanging 
drop containing the organisms to be inoculated should be put 
under the cover glass, and near it a hanging drop of sterile water 
should be placed. These should be as near as possible to the 
cells to be inoculated. If a paraffin barrier is used, they may be 
placed just outside of it. Lines should be drawn on the cover, 
preferably with India ink, to serve as guides to different parts 
of the preparation. When all is ready, the isolating chamber is 
placed on the stage and the outer edge brought to focus under 
the low power. 


* For a description of the method of cultivating fish molds for inoculation, 
see This Journal, Sec. B (1918), 8, 373. 


350 The Philippine Journal of Science 1914 


The microburner is placed to the right of the microscope, 
as usual, and to the right of the microburner and near the edge 
of the table is placed a large crystallizing dish about 5 centi- 
meters in height, full to the brim with water containing ice. 
To the left of the microscope is placed a special apparatus for 
regulating the temperature of the loop. 

This apparatus (fig. 18) consists of the brass tube T, to the 
lower part of which is attached the brass cup C. This cup is 
elliptical in horizontal section with a long diameter of about 
8 centimeters and a short one of about 4.5 centimeters. The 
tube J is held in the sleeve S, which is attached to the arm of 
a simple microscope provided with a ratchet and pinion R. 
By means of the ratchet and pinion, the sleeve and tube may 
be raised and lowered through about 5 centimeters. The tube 
may be raised or lowered in the sleeve; a joint at J allows the 
cup to be swung aside. A Petri dish is placed beneath the 
cup to receive waste water. The cup must be filled with water 
containing snow or small pieces of ice. 

The pipette is now to be filled with mercury to the tip of 
the capillary. Heat the loop gently in a flame, and immerse 
the point of the capillary in mercury the moment all air is 
expelled. With the tip still in the mercury, immerse about two- 
thirds of the loop into the crystallizing dish containing ice 
water and hold it in this position for a quarter of a minute or 
so. Now remove the tip from the mercury, and immerse the 
loop still farther into the ice water. The mercury in the capil- 
lary will retreat 4 or 5 centimeters from the end. Make the 
pipette point immediately, following as nearly as possible the 
same technique as in the case of isolating pipettes. The loop 
should remain in the ice water, and the arm next to the loop 
may rest on the edge of the crystallizing dish. The arm should 
be kept nearly horizontal and held with the right hand at a 
point near the beginning of the capillary. The forceps, held in 
the left hand, grasp the capillary. The microburner must be 
slipped to a convenient distance from the dish. One may draw 
the pipette more conveniently if seated low enough to bring the 
eye near the level of the flame. 

For this work it is necessary to have a closed tip, coming 
rather abruptly to a very fine point, a (fig. 19). A point more 
tapering can be used as 0b (fig. 19), but such points are more 
liable to become clogged with mercury, and do not allow of the 
control of the dosage as well as the kind illustrated. One will 
save time eventually by making a point of the proper form. 
After a suitable tip is drawn, it should be turned at right angles 


dol 


Barber: The Pipette Method 


IX, B, 4 


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pus joyoyer “yy “doo, oy} JO oin}Bredui9, oy} Buryepnsoa 10Z snjyersdds oy} YIIM UolzIsod ul o}JodId UOlyeE[NoOOUL oY T, “ST “dW 


CoM 


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Vwuuly 


352 The Philippine Journal of Science 1914 


or slightly less. Immediately before turning the point, dip the 
loop wholly in the ice water. Unless there is a partial vacuum 
in the capillary at the moment of bending, the heated air may 
expand and burst through the softened glass. One may inspect 
the point with a hand lens or under the low power before turning 
it, but the loop should not be left long out of the ice water while 
the point is still sealed. The pipette is now ready for use and 
is to be adjusted in the holder. As soon as the tip is brought 
near the edge of the cover glass, the cup C should be brought 
into place and raised so as to immerse the loop in the ice water. 
The mercury is under great pressure at room temperature, and 
may break the pipette if it becomes warm while the tip is still 
sealed. The final adjustments can be made with the loop in the 
cup. The cup should be so adjusted in the sleeve that when it 

is racked down to its fullest 


‘ ’ extent the bottom of the loop 
will just clear the water in the 
cup. 


The point should now be ad- 
justed to the center of the high- 
power field. The tip should be 
broken off under the high power, 
and great care must be taken 
to avoid breaking it prema- 
turely. It is best to bring the 
tip into the hanging drop near 
its edge, but at a little distance 
Fic. 19. Various types of tips used in cell below the cover. If left too 
(From The J cutuinctaenene Diseases. ) near the glass, it may be broken 

by a slight bending of the cover 
during the focusing with the oil immersion. Sometimes it is 
more convenient to adjust it under the low power just beneath a 
small droplet of water, or any object easily recognized, and then 
lower it slightly. The object is then focused under the oil- 
immersion lens, and the tip is raised into view. The inoculation 
should be done under high power. An oil-immersion lens of 
deep focus is by far the best for this work. 

The tip, still closed, is now in focus under the high power. 
The end should be opened by pressing it very gently against the 
cover and moving the mechanical stage slightly. By varying 
the pressure on the cover glass, the amount broken off and the 
size of the opening can be regulated. The size must depend on 
the substance to be inoculated. If liquids, only the smallest 


Ba” Barber: The Pipette Method 353 


possible opening is made; if bacteria, a larger opening is needed; 
and if yeasts or spores of fungi, one still larger. As a rule, 
the opening should be as small as the nature of the work will 
permit. If desired, a point with an opening scarcely exceeding 
1 micron in diameter may be made. To determine whether a 
fine point is open or not, one has only to touch it to a droplet 
of condensed moisture. If the point is open, the liquid will 
instantly enter by capillary. 

With the tip in the hanging drop of water, lower the cup and 
allow the mercury to expand sufficiently to expel the air from 
the pipette. As the pipette was supplied with mercury with 
the loop partially cooled in ice water, the room temperature 
will furnish sufficient heat for the expansion of the mercury. 
When mercury appears at the tip, raise the cup enough to stop 
the pressure. If some mercury comes out, no harm is done, 
except when in very tapering pipettes a plug of mercury some- 
times clogs the tip. Now bring the tip into the hanging drop 
of bacteria or other substance to be inoculated. If the mercury 
column has retreated, apply a little pressure to bring it to the 
tip again. Raise the cup until sufficient negative pressure has 
formed to draw in the dose required. The size of the dose can 
be regulated roughly by the extent of the fall of the mercury 
column. Then lower the pipette and, as quickly as possible, 
move to the organism to be inoculated. Bring the tip under it, 
and raise the point until it penetrates the cell wall and reaches 
the vacuole or the part of the cell desired. Now apply pressure 
gradually until the dose is seen to come out and enter the cell. 
Then stop the pressure at once by raising the cup containing 
the ice water until it surrounds the loop. 

In order that the dosage may be regulated and that mercury 
may not be driven into the cell, the tip of the pipette should be 
unobstructed when it pierces the cell wall. Substances may 
successfully be injected in spite of a clogged tip, but so much 
force is often required to overcome the obstacle that injection 
takes place with a rush and mercury is apt to follow the dose. 
Further, both for the regulation of the dosage and the prevention 
of clogging, pressure in the pipette should be nearly in equili- 
brium at the time of the entrance of the point within the cell. 
Positive pressure may not be a disadvantage if one does not care 
to avoid ejecting some of the contents of the pipette outside of 
the cell, but a too strong negative pressure causes the pipette 
to take up liquid surrounding the cell and increases the danger 
of clogging when the point enters the protoplasm of the cell. 


304 The Philippine Journal of Science. 3914 


A clogged pipette may often be opened by gently scratching it 
on the surface of the cover glass. Sometimes it is necessary to 
enlarge the opening somewhat before it can be used. 

The cup must be well supplied with ice-cold water, and the hand 
should be on the adjustment R, ready to bring it up quickly. 
In case one is not particular as to the size of the dose, an excess 
of bacteria may be drawn into the pipette and the pressure 
stopped when some of the bacteria have come out. 

After the dose is in, the pressure is at once diminished and 
the pipette withdrawn. With very fine tips the point may be 
drawn out with little or no loss of cell contents. With larger 
tips it is often necessary to withdraw the tip very slowly until 


the plug of protoplasm, which usually forms at the point of . 


inoculation, is sufficiently firm to close the opening. In large 
openings the dose is sometimes drawn back into the pipette on 
reducing the pressure. Here one may introduce the tip just 
through the layer of protoplasm and wait until a mound of 
protoplasm has been formed over it before injecting. This will, 
in many cases, act as a valve in closing the opening after the 
dose passes out, so that the tip may be withdrawn safely. 

After inoculation, the cover may be removed and placed in a 
moist chamber or sealed on a hollow slide. If it is desired to 
keep the substance to be inoculated some distance from the 
organism, it may be placed on another cover, the dose taken 
in, the tip lowered, and the covers changed for inoculation. 

A filament grown beneath the surface of a hanging drop of 
agar may be inoculated, but in that case it is well to have the 
tip nearly at right angles with the capillary, else it may bend 
too much in entering the agar. 

In working with pure cultures, it is sometimes necessary to 
avoid getting any of the fluid or culture to be injected into the 
medium outside of the filament. Here, after filling the pipette, 
it may be quickly washed in a sterile hanging drop of water 
or agar and then brought to the cell. It is obvious that a tip 
with a relatively large opening may, if the pressure is negative, 
draw in the washing fluid or air during transfer, or, if positive, 
may eject prematurely part or all of the dose. So with such 
points, it is best to have the pressure as nearly neutral as pos- 
sible. Fine openings give much less trouble in this respect. 

If some bacteria are forced out into the surrounding medium, 
it is sometimes possible to remove them with an ordinary pipette 
attached to a rubber tube, such as is used in ordinary isolation. 
These pipettes, either under or apart from the microscope, may 


IX, B, 4 Barber: The Pipette Method 855 


be used to remove the medium surrounding the infected filament 
and to substitute any other during the course of an experiment. 

If it is found that there is not enough pressure to eject the 
dose, one may move a Bunsen burner or electric light to a point 
a few centimeters from the loop. Or a rubber bag containing 
warm water may be adjusted by means of a bent wire holder 
to the sleeve of the pressure apparatus, so that the loop in 
rising will come into contact with it. This is rarely necessary 
if the loop is cooled sufficiently below room temperature while 
the capillary is being filled with mercury. The pipette may be 
so supplied with mercury that it will be under negative pressure 
at room temperature. In that case, warm water is kept in the 
cup in place of the cold, and the necessary expansion of 
the mercury is obtained by immersing the loop in it. But the 
arrangement first described best meets the main requirement— 
the possibility of gradually applying pressure and of stopping 
it quickly. 

If a volatile liquid is to be injected, or if large or repeated 
doses of the same substance are to be used, one may fill part of 
the capillary with the substance to be inoculated before making 
the injecting point, but in most cases one can better regulate the 
dosage by filling from the point. If very small doses are 
to be injected, one may keep the top of the mercury column in 
view after supplying it with the dose. The cell is pierced, pres- 
sure is applied, and the rising of the mercury column to the tip 
shows that the dose has come out. Focusing on the mercury 
column may be facilitated by piercing the cell obliquely with a 
pipette bent at less than right angles, instead of from directly 
below. With larger doses, the top of the mercury column is 
usually below the reach of the lens. Here, one can focus as far 
down as possible in the lumen of the pipette after its introduc- 
tion into the cell and stop the pressure on the appearance of 
the mercury column. If pipettes of the form represented in fig. 
19, a, are used, much more force is required to expel the mercury 
than to bring it to the tip, so that one has time to stop the 
pressure before any mercury can come out. If it is desired to 
remove cell contents, the retreat of the mercury column indicates 
that the contents are being drawn into the pipette. 

Small doses may be measured by estimating the cubic contents 
of the pipette between the top of the mercury column and the 
tip. Larger ones may be estimated by expelling the dose on 
the cover glass and measuring the droplet expelled by compar- 

1284434 


356 The Philippine Journal of Science 1914 


ing it with one of known volume. (For methods of estimating 
the size of hanging drops, see page 342.) The liquid is then 
drawn into the pipette again. If a definite number of bacteria 
are to be inoculated, they may first be isolated in a droplet of 
fluid and the whole droplet inoculated. 

If the substance to be inoculated forms a precipitate with the 
mercury, a quantity of water or of some indifferent oil, sufficient 
to separate the substance to be injected from the mercury, may 
first be drawn into the pipette. 

One should have the cell to be injected well located before 
filling the pipette and as near to the filling place as possible, © 
so that little time will be lost between charging and injecting. 
This is the more necessary where it is advisable to keep the pres- 
sure in the pipette in equilibrium after filling. Lines may be 
drawn on the cover glass to serve as guides, the cell may be 
located by means of the vernier on the mechanical stage, or the 
droplet of the substance to be injected may be placed just in line 
with the cell so that only one movement of the mechanical stage 
is necessary in passing from the one to the other. 

The penetration of the plant cells thus far experimented on 
is easy, if the tip is made fine enough and if the capillary back 
of it is thick enough to give the pipette the necessary stiffness. 
If the point of the capillary is too pliable, there may be difficulty 
in penetrating the cell; if too blunt, there is danger of tearing 
the cell wall. Sometimes in tough-walled plants there may 
be some difficulty in piercing the wall without using so much 
pressure that the pipette, on entering, will penetrate too far. 
In such cases, one may often obviate the difficulty by pressing 
the tip against the wall, and then, by moving the mechanical 
stage gently, bore a hole in the wall. 

In the multinuclear cells of the fungus group Saprolegniacese 
and of Nitella and Vaucheria among the alge, the cell wall has 
‘been pierced, different substances injected, and the pipette 
withdrawn with little or no apparent injury to the cells, as 
judged by the movements of the protoplasm and the subse- 
quent behavior of the cell. With some mononuclear alge 
cells, as Spirogyra, the cell appears to be more sensitive to 
injury. Few animals have thus far been experimented on with 
the injection apparatus. Mercury has been injected into 
rotifers, and other substances have been injected into Para- 
mecium. The larve of a gnat have been infected with bacteria, 
and larve of Stegomyia have been inoculated with defibrinated 


1X, B, 4 Barber: The Pipette Method 357 


human blood. Some work has been done by me on the effect of 
poisons and other substances injected into the cells of Nitella.1® 

In making a new capillary point, the pipette may be removed 
from the holder, the old point broken off, and a new one made 
from the same capillary. When the capillary is used up, a new 
one may be made from the end of the straight portion of the 
pipette, and the process may be continued until all is used back 
to the loop. If a capillary point remains in good condition after 
use, the pipette may be removed from the holder and kept for 
later use; but it must be placed in a refrigerator, else the expand-. 
ing mercury is apt to burst it. 

In certain kinds of work it is of advantage to use two pipettes 
simultaneously. This makes necessary the use of a second 
holder for another pipette (fig. 15). With this modification, 
one may inject two different substances into the same cell 
at the same time, or by varying the pressure inject with one 
pipette and withdraw with the other. One pipette may be 
used simply as a probe or dissecting instrument, or it may be 
attached to a rubber tube and used as an isolating pipette while 
the other is arranged for injection. In one experiment, a 
rotifer was held by the blunted point of one pipette while mer- 
cury was injected into the body with the other. Points may be 
made fine almost to invisibility, with sufficient stiffness for 
piercing the wall or even the membrane of the nucleus of a cell. 

It has seemed to me that this inoculation technique in its 
different forms may assist in the solution of various problems 
in the biology of microscopical plants and animals. The in- 
troduction of foods, poisons, stains, and fixatives is made pos- 
sible, and cells may be probed or dissected under high power— 
methods which may be of use in the study of the structure, 
chemistry, and physiology of cells. Finally, materials may be 
withdrawn from one cell and injected into another, and it is 
possible that investigations on fertilization and heredity may 
be extended by this technique. 

In conclusion, it may be stated that this paper aims to give 
the principle on which the pipette technique is founded and 
only a part of its applications in detail. Experienced laboratory 
workers, especially in fields less familiar to me, may make new 
applications of the method or changes in the details of those de- 
scribed here. I believe that the method may be much further 


* Journ. Infect. Dis. (1911), 9, 117. 


358 The Philippine Journal of Science 


developed in some directions, and may even find some applica- 
tions in fields other than those of biology. I shall gladly wel- 
come any suggestions as to the improvement of the technique 
as described here or of its application to other fields of research. 

I take pleasure in acknowledging the codperation of my 
former students, Dr. A. W. Sellards, Dr. Montrose Burrows, 
and Dr. Frederick Hecker, in developing this technique. I am 
also under obligations to Professor E. F. Stimpson, of the physics 
department of the University of Kansas, and Mr. C. W. White, 
. instrument maker of the University of Kansas, for assistance 
in working out the mechanical details of the pipette holder. 


| 


ILLUSTRATIONS 


(Photomicrographs by Charles Martin) 
PLATE I 


Fic.1. Spores of Aspergillus. Two isolated and two in one droplet. 
Freshly isolated and unstained. The amount of condensed 
moisture proper for isolation is shown in this figure. 

2. Spore of Bacillus subtilis isolated in a drop of liquid. Freshly 
isolated and unstained. 

3 to 7, inclusive. Bacterium coli commune. This series was obtained 
as follows: One organism was isolated in broth plus a little serum. 
When it was divided into two, a daughter cell was picked up and 
transferred to a new droplet. When this had divided, a cell was 
transferred to a third droplet. After four successive transfers, 
a series of 5 droplets was obtained containing respectively 1, 2, 
4, 8, and 16 bacilli—the offspring of the individuals left behind at 
each transfer. The cover glass was dried and the bacilli stained 
on the cover without the loss of a single one from the series. 
The diameter of the droplets was, on the average, about equal 
to the length of the chain of 8. 


PLATE II 


Fig. 1. Larva of Culex injected with mercury by means of the inoculation 
pipette. 

2. Antenna of the larva of Culex shown in fig. 1. This was dissected 
away, and after removal injected with mercury with the in- 
oculation pipette. The point of the pipette was introduced 
between the spherical and the elongated masses of mercury. 


TEXT FIGURES 


(Text figures 9 and 19 by courtesy of The Journal of Infectious Diseases. Drawings of other 

text figures by Moscaira and Espinosa) 

Fic.1. Pipette holder containing a pipette (pip). cl, clamp by which 
the holder is fastened to the metal plate pb; ud, up-and-down 
adjustment governed by screw s; rl, right-and-left adjustment 
governed by screw s’; g, groove in which the pipette is held by 
plate tp; ic, isolating chamber; H, hood of pasteboard for pro- 
tecting the end of the isolation chamber. 

2.Top of pipette holder. rl, right-and left adjustment governed by 
screw s’; g, groove in which the pipette is held by plate tp and 
set screw Ss. 

8. Isolating chamber. vp, lining of blotting paper; s, glass strip for 
retaining water in bottom. 

4. Cover glass marked with cross lines of India ink, and supplied 
with hanging drops of sterile fiuid. a, drop to which the 
bacteria to be isolated are added. 

359 


360 


The Philippine Journal of Science 


Fic. 5. Method of making the capillary pipette. 6, microburner. 
6. a, completed pipette. 06, c, and d, various sorts of points. 
7. A dissecting microscope used as a pipette holder. 
8. A cover glass with droplets arranged for an extended series of 


isolations. c, a droplet shown on a larger scale with a small 
bacterium-containing droplet beside it. 


9. Two-movement pipette holder, pipette and isolation chamber ar- 


19. 


ranged to illustrate method of transferring single isolated bacteria 
by means of a perforated mica plate. ms, mica plate; cv, small 
cover glass over perforation; x, large cover glass; p, pipette; g, 
up-and-down adjustment of holder moved by screw f/f; 7, to-and-fro 
movement regulated by screw s. 


. A capillary tube in position for taking up isolated organisms. 
. A pipette constructed for use above a barrier of blotting paper. 
. Special isolating chamber in longitudinal section. The chamber 


6 is closed at the side by the trap ¢ and partition p. 


. A special pipette in position for isolating organisms from the 


bottom of a Petri dish. p and pa, metal plates screwed to the 
stage for the attachment of the pipette holder. The complete 
pipette holder is used, but only a portion of it is shown in the 
figure. 


. Special pipette for measuring small quantitics of liquid. 
. Microscope with two pipette holders, each containing a pipette 


attached to the stage by means of metal plates. Seen from the 
back. A, a three-movement, and B, a two-movement holder. 
tf, adjustment, governed by screw s”, for moving the pipette to 
and from the observer. The other letters correspond to those 
shown in figs. 1 and 2. 


. Apparatus for attracting certain motile bacteria into pipettes. 
. Pipette for injecting substances into living cells. Completed. 
. The inoculation pipette in position with the apparatus for regulat- 


ing the temperature of the loop. R, ratchet and pinion of a 

simple microscope; S, sleeve of metal fastened to the lens holder 

which is jointed to the microscope at J; T, hollow metal tube 

inserted in the sleeve and bearing at its base the cup C. 
Various types of tips used in cell injection. 


| 
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BARBER: THE PIPETTE METHOD. ] [Puin. Journ. Scr., IX, B. No. 4. 


Fig. 5. Fig, 6. Fig. 7. 


Fig. 1. Spores of Aspergillus, freshly isolated and unstained. Fig. 2. Spore of Bacillus subtilis, 
freshly isolated and unstained. Figs. 3-7. Bacterium coli commune. 


PLATE I. 


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Fig. 1. Larva of Culex injected with mercury by means of the 
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Fig. 2. Antenna of the larva of Culex shown in Fig. 1. 


PLATE Il. 


NOTES ON THE DIAGNOSIS OF ASIATIC CHOLERA AT AUTOPSY 


By B. C. CROWELL 


(From the Department of Pathology and Bacteriology, University of 
the Philippines, and the Biological Laboratory, 
Bureau of Science, Manila, P. I.) 


Without attempting to review the entire pathological anatomy 
of Asiatic cholera, it may be of advantage to note some facts 
obtained in our recent experience with 92 cases of the disease, 
which may be useful in diagnosis at the autopsy table. 

The autopsies were performed largely as a diagnostic measure 
to assist: the Bureau of Health in locating cases of the disease. 
This naturally entailed the examination of a much larger number 
of bodies than appears in this series. Ninety-one and three- 
tenths per cent of the series occurred in Filipinos, the remainder 
occurring in Japanese, Chinese, Americans, and Spaniards. Sixty 
of the cases were males and 32 females. The ages ranged from 
17 days to 99 years, the greatest number occurring between the 
ages of 21 and 25 years. 

The work of the pathologist has gone hand-in-hand with that 
of the bacteriologist in the recent epidemic, and a loop of small 
intestine has been submitted to the bacteriologist from every 
autopsy that has been performed during the period of prevalence 
of the disease. 

As this disease does not exist without the presence of the 
cholera vibrio, the bacteriologist is naturally considered the court 
of last appeal in the diagnosis of the disease. In the present 
series, the number of cases in which there has been disagreement 
between the pathologist and the bacteriologist were comparatively 
so few, that these cases have been very instructive, and have 
served to emphasize certain facts which should be more widely 
recognized. 

The autopsy findings in cases of Asiatic cholera vary according 
to (1) the stage of the disease at which the patient died, (2) 
whether or not the patient had received treatment, and (3) the 
presence of other associated diseases. The disease is notably 
so severe in its action that a large number of persons previously - 
of great bodily strength and vigor succumb to its influence. On 
the other hand, cholera is also one of the severe intercurrent in- 

361 


362 The Philippine Journal of Science 1914 


fections which ends the life of those who were previously the 
victims of a more chronic disease. Forty-eight of our cases had 
no marked evidence of disease other than cholera, while 17 had 
also lesions of tuberculosis, and smaller numbers had syphilis, 
marked arteriosclerosis, beriberi, lymphatic leukemia, strangu- 
lated hernia, suppurative hepatitis, Banti’s disease, status lym- 
phaticus, bronchopneumonia, and splenomegaly. There were 4 
full-term pregnant women in the series, one of whom aborted. 

The post-mortem appearances, like the clinical manifestations 
of the disease, are due to the loss of fluid from the tissues and to 
the action of the toxin of the cholera vibrio on the tissues. 
These appearances may be modified by treatment or associated 
disease. 

Rigor mortis.—Rigor mortis appears early, and is stronger 
than that following any other known disease. The tissues of 
the hands and feet especially appear shrunken so that the bones 
and interosseous spaces are prominent. The nails are blue, and 
the fingers are usually strongly flexed. The soft tissues covering 
the skeleton feel very dry, and have a doughy consistence. 
Frequently evidences of the diarrhea are present about 
the buttocks. The abdominal muscles are usually rigid, and 
present the dull, homogeneous, waxy appearance of Zenker’s 
degeneration. 

On section the subcutaneous tissues are dry, and the peritoneum 
and pleura are usually devoid of fluid and are sticky. In no case 
has the pericardium been devoid of fluid or sticky. In cases 
that have been treated by the administration of large quantities 
of fluid, this dryness of the tissues may not be a marked feature. 
The small intestine is usually somewhat dilated, and presents a 
dry, sticky, rosy, or pink serosa, sometimes with marked injec- 
tion of the vessels, standing out in marked contrast to the pallor 
of the peritoneum covering the large intestine, stomach, and 
parietes. In other cases the serosa of the small intestine has a 
slaty gray color. The large intestine is usually contracted. The 
liver is frequently retracted above the costal margin. The lungs 
are as a rule poorly inflated, and the pleura is exceedingly dry. 
In cases which have lost much fluid, the cut surfaces of the lungs 
are red but very dry, while in the very early cases, or in those 
which have received fluid, congestion and cedema of the lungs 
may be marked. The mucosa of the bronchi is usually reddened. 
Pleural ecchymoses are not infrequent. 

The heart.—The heart presents evidence of degeneration of 
the muscle and sometimes a dilated right ventricle. The blood 
within the heart is poorly coagulated and is very dark. This is 


ne 


IX, B, 4 Crowell: Diagnosis of Asiatic Cholera 363 


true also of the blood in all parts of the body, and its total volume 
may be very much diminished. In a fair percentage of the cases, 
however, a mixed coagulum is present in the right side of the 
heart. No acute endocardial lesions have been encountered. 
Subendocardial hemorrhages sometimes occur, while epicardial 
ecchymoses are the most frequent hemorrhages found in the 
body; these are, however, not present in a large proportion of 
the cases. 

The spleen.—The spleen is usually much diminished in size 
and is flabby, with wrinkled capsule. The cut surface is dry, 
smooth, and dark red in color. 

The kidneys.—The kidneys present either the evidences of an 
acute parenchymatous degeneration or hemorrhagic nephritis. 
In the former case, they are pale and dry and the cortex appears 
pale, yellowish gray, compact, opaque, and relatively rather wide; 
the vascular markings in the cortex are not usually readily visible. 
In the kidneys of the hemorrhagic type, the organ is more 
swollen, much darker in color, and the vessels are more promi- 
nent, both on the surface and within the organ. 

The urinary bladder—tThe condition of the urinary bladder 
is a matter of some interest. In 60 of our cases it was firmly 
contracted and empty, and 11.6 per cent of these cases had been 
treated. In 18 cases there was a small amount of urine in the 
bladder, often only 2 or 3 cubic centimeters, and 77 per cent of 
these cases had been treated. In the remaining 14 cases no 
record was kept of the condition of the bladder. I know of no 
other epidemic disease in which a firmly contracted empty 
bladder is so constant a finding. The explanation probably lies 
in (1) the lack of urine, due to (a) loss of fluid through the 
intestine and (b) specific toxic action on the renal epithelium, 
and (2) strong cadaveric rigor of the bladder muscle. 

The liver.—The liver is diminished in size as a rule, from loss 
of fluid. It frequently appears dark and congested, probably 
from concentration of the blood, and presents evidences of acute 
parenchymatous degeneration from the action of the toxins of 
the cholera vibrio. 

The gall bladder.The gall bladder may present the condition 
of hydrops or acute inflammation. This has been very infrequent 
in our series, only 3 cases of marked inflammation having been 
recognized. 

The stomach.—The stomach frequently contains a moderate 
amount of brownish fluid; a catarrhal or hemorrhagic inflamma- 
tion may be present. In one case a pseudomembranous or 
gangrenous gastritis was present. 


364 The Philippine Journal of Science 1914 


The mesenteric and mesocolic glands usually show no change. 

The brain.—The brain usually shows no change other than 
injection of the meningeal vessels. In about one-third of the 
brains examined, the note is made that the meninges are dry, 
while in two cases associated with arteriosclerosis there was pial 
oedema. 

The intestine.—The intestine is the organ about which the 
greatest interest is centered, but sometimes the findings here 
are scarcely sufficient to justify a diagnosis without taking the 
other features into consideration. The condition of the serosa 
has already been described. The contents of the intestine vary 
somewhat in character and amount according to the stage of the 
disease and the amount of fluid which has been administered 
in treatment. Practically never is any solid or formed fecal 
matter present. The usual finding is a smaller or larger amount 
of fluid, usually pale, with an abundant admixture of mucus and 
some epithelial flakes, This is sometimes streaked with blood, 
sometimes deeply bile stained, and sometimes brown in color. 
In children it may be green. The amount varies from 2 to 3 
liters down to a very small amount of mucus without fluid. 

The lesion of the intestine itself is essentially an acute ca- 
tarrhal enteritis. The mucosa, especially of the lower portion of 
the ileum, appears pale, smooth, glistening, and almost trans- 
lucent; the lymphoid tissue appears prominent, especially that 
of the solitary follicles. This same appearance may be present 
in the colon and stomach. Sometimes the mucosa, especially 
over and around the lymphoid tissue, appears very red, and 
occasionally there are actual hemorrhages in these areas. In 
2 cases of the series an actual pseudomembranous enteritis and 
colitis existed. Without having made definite percentages, I 
should say that animal parasites were no less frequent than in 
our usual routine autopsies. The conditions described above 
may not be very marked, but when taken into account with the 
findings in other organs usually suffice for a diagnosis. The 
lack of formed fecal matter is extremely constant. The difficulty 
is enhanced when other intestinal diseases are present in the 
ileum, as for example tuberculosis and status lymphaticus. Even 
where these coexist, attention to the above features will at least 
arouse suspicion if not make diagnosis possible. 

To recapitulate, it may be said that, while probably no one 
anatomical feature is constant, the following features are the 
ones on which a diagnosis is chiefly based: 

Acute catarrhal enteritis associated with (1) cyanotic finger 
nails, (2) dry tissues, (3) oligemia, (4) dry and sticky peri- 


IX, B, 4 Crowell: Diagnosis of Asiatic Cholera 365 


toneum with pink serosa of ileum, (5) contracted and empty 
urinary bladder, (6) shrunken, dry spleen and liver, (7) acute 
degeneration of parenchymatous organs, (8) poorly coagulated 
blood, (9) absence of formed feces, (10) presence of rice-water 
intestinal content, and (11) prominence of lymphoid tissue in the 
ileum. 

After all the autopsies performed, we have placed ourselves 
on record without knowledge of the bacteriological findings, 
except in a very few cases. The cases were classified as (1) 
cholera, (2) not cholera, (8) probable cholera, or (4) possible 
cholera. Comparison of the anatomical and bacteriological find- 
ings shows that 5 cases anatomically negative were bacteriolog- 
ically positive. Eleven of the probable cases were bacteriologi- 
cally positive, while only 1 was negative. Three of the possible 
cases were positive, while 4 were negative. For a correct inter- 
pretation of these findings, it should be borne in mind that the 
mere presence of cholera vibrios does not mean that the patient 
was suffering from cholera. In 87 cases there was no difference 
between the two findings. The cases which were found bacte- 
riologically positive in which the pathologist had committed 
himself to the diagnosis of not cholera were (1) beriberi, (2) 
leukemia and pulmonary tuberculosis, (3) bronchopneumonia 
and post-mortem decomposition in a 17-day old child, (4) acute 
peritonitis from strangulated inguinal hernia, and (5) general- 
ized tuberculosis. 

The other cases in which the bacteriological findings were 
negative whereas they had been called probable or possible 
cholera by the pathologist were cases of acute enteritis, and 
the suspicion was and is deemed justifiable at a time when 
cholera is epidemic. 


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TYPHOID FEVER IN THE PHILIPPINES * 


By PERPETUO GUTIERREZ 


(From the Department of Medicine, University of the Philippines, and the 
Philippine General Hospital) 


During recent years a number of observers have contributed 
to our knowledge of typhoid fever as it occurs among foreigners 
in the tropics. Very little work has been published regarding 
the incidence, clinical types, and mortality of the disease among 
native inhabitants of warm countries. 

The material for this paper is based upon the study of 125 
cases of typhoid fever in Filipinos, treated in the Philippine 
General Hospital during a period of a little less than two years. 
During this time, there were 137 cases of typhoid admitted to 
the hospital, of which 6 were Americans, 3 Portuguese, 1 English, 
2 Japanese, and the remainder Filipinos. 

The importance of recognizing the incidence index of typhoid 


- fever in this country cannot be overestimated, because once this 


infection gains a foothold it will be difficult to eradicate, on 
account of the peculiar environment and the poor hygienic con- 
dition under which most of the people live. 

There seems to be considerable difference of opinion regarding 
the local distribution and prevalence of the disease. Chamber- 
lain, Nichols, La Garde, and others believe it to be prevalent 
throughout the Archipelago. Heiser, on the other hand, claims 
that the incidence was very low prior to 1910. Whether or 
not the disease is on the increase, I am not in a position to say. 
Certainly more cases came to the hospital this year than in pre- 
vious years. In 1911 there were 23 cases, or 0.66 per cent, of 
typhoid out of a total of 3,461 admissions. During the fiscal 
year 1912 there were 39 cases, or 0.53 per cent, of typhoid 
patients out of a total of 7,252 admissions. During these two 
years, therefore, the number of typhoid patients admitted shows 
a fairly uniform relationship to the total admissions. Begin- 
ning with the fiscal year 1913 and up to the present time, there 
have been 98 cases of typhoid out of a total of 6,300 admissions. 
This marked increase may be due, in part, to the fact that people 
are getting more used to hospitalization, but this cannot explain 
all the factors concerned in the increase, and if we may draw 
conclusions from the statistics of the hospital typhoid fever is 
more prevalent now than it was during previous years. 


“Read at the meeting of the Colegio Médico-Farmacettico de Filipinas, 
June 21, 1913. 
367 


LOR RS TO Oy en ee ae 


368 The Philippine Journal of Science 1914 


Seasonal incidence is a characteristic of typhoid in Europe 
and the United States where the greatest prevalence is seen 
during the autumn months, continuing into the winter, and 
declining during the spring months. Examination of the fol- 
lowing table indicates the same seasonal distribution in this 
country that is encountered in temperate climates. However, 
the variation is not so marked and the series of cases is too small 
to justify definite conclusion. 


TABLE I.—Seasonal incidence of typhoid in the Philippine Islands. 


Number of cases. 
Month. —______———_| Total. 
1912. 1913.4 


JQnuarys) 23/25 ke Ve a oe ed 1 9 
Webroary22i 402.2 ee ee a ae a ae ee 3 uf 
Manele 4.5) Sore 2 eee ee Bes PEE ee ee ee ee eee ee 4 5 
April 22623518 St ee Ss ee ee ee ee 5 6 
Way 30S oS Eo Ee ar a ees 7 8 
UNS 226 Se eee EE Sef  SU ES IS eee ee ee 2 8 
BUMS ALOE chee Se Se Oe Se oe Rs Cee ee ae 6 6 | 
PAICMIBE > 580-4 hac tee ek on gd ee Soe 2 8 
Sep tent Der = tee ee So a ae Se eae a ie ee il 
October tesa be = AONE Soe eR ee eee ee ee Se eee 2 15 
|: November 2:2. 1052... yl et ede A i eee 2 11 
December cio. 522 335 5 Bee ee ee ERE eR ee Ek el 5 9 


* Figures for the fiscal year 1913 are incomplete. The last case in this series was on May 28. 


Sex.—The sex incidence of 85 males and 40 females out of a 
total of 125 cases corresponds closely enough with sex statistics 
of this disease in other countries. 

Age.—tThe distribution of the disease by ages is shown in the 
following table. 


TABLE II.—Age incidence of typhoid in the Philippines. 


Age. 
| | O Total. 
10 11 to 14/15 to 25/26 to 8031 to 40] ~jy* 
years. | years. | years. | years. | years. | )oa16. 
Miales)...° 32:2. se 2 eee ee Sa 3 4 53 13 10 2 85 
Memaler= i... 2 oe ee cee es 2 3 29 3 2 uf 40 
Total 2.2 oe re et 12 82 28 3 126 
Percentage. oo es 9.6 65.6 22.4 PAR ER eesinesy) | 
Rogers... - 5325653 ee eee 41.6 47.23 bb Bh Reese er 
Curschmann: 
11.02 | 48.68 
9.59} 49.40 


——— 


IX, B, 4 Gutierrez: Typhoid Fever in the Philippines 369 


Rogers calls attention to the frequency of typhoid in children 
in India. In his Calcutta series 41.67 per cent of his cases 
occurred in patients under 15 years of age. Manson also calls 
attention to the frequency of typhoid among oriental children, 
but Nichols, writing from the Philippines, states that only 9.6 
per cent of the total cases occurred in patients under 15 years 
of age. In examining the table, it is seen that the percentage 
of infection, according to ages, in my series corresponds fairly 
well with statistics from other countries. 


CLINICAL DESCRIPTION 


The following discussion of the clinical picture of typhoid as 
seen in this country is based on the study of 125 cases treated 
during a period of a little less than two years. Diagnoses of 
about 90 per cent of these cases were verified by blood culture 
or serum reaction. Of the remaining cases the diagnosis was 
based upon the course of the disease, the fever, low leucocyte 
count, enlarged spleen, etc., and in some of these the diagnosis 
was verified by autopsy. It is fully realized that the number 
of cases studied is too small to justify conclusions regarding the 
incidence of rare complications, but the evidence seems to be 
useful in analyzing the most important features of the disease 
as it is seen in the Philippine Islands. 

The disease, in a general way, presents essentially the same 
symptoms in the tropics that it does in the temperate climates; 
yet there are a few exceptions which stand out prominently 
before the clinician. The mortality seems to be higher while the 
fever is lower, and the duration of the fever at first sight would 
seem much shorter than that encountered in other countries. 
The temperature curve is less frequently characteristic than 
it is in the temperate climates. 

The course and type of temperature.—The typical typhoid 
temperature of the temperate climates is divided into three 
stages: the steplike rise at the outset, the evening temperature 
always higher than that of the evening before, which lasts for 
a period of from three to five days, after which the stage of 
continued fever or fastigium is recognized. During this period 
the temperature ranges from 39°.5 to 40°.5 C. with but very 
slight remissions. This stage lasts from a few days to three 
or four weeks, and ushers in the third stage, or the stage of 
decline, during which the remissions become more and more 
marked until the normal is reached. This typical temperature 
in typhoid is very seldom encountered in the Philippine Islands. 
Indeed, such a classical temperature curve is so rare that when 


370 The Philippine Journal of Science 1914 


it does occur it is considered to be of grave importance in prog- 
nosis as well as in elucidating the diagnosis. In most of our 
cases we have come to rely more upon the relation between tem- 
perature and pulse in making a diagnosis than upon the tem- 
perature itself. 

The onset of the fever usually is so irregular that the sugges- 
tion of the possibility of typhoid does not occur until the disease 
is well advanced. The most frequent symptoms during this 
period in the series of cases under discussion were headache, 96; 
general pain all over the body, 25; chilly sensation, 29; malaise, 
24; abdominal pain, 22; pain in the right iliac region, 3; epis- 
taxis, 1. Headache is the most constant of all symptoms, often 
intense and frequently lasting well into the second week of the 
disease. Very frequently the patients state that the onset was 
sudden, preceded by a chill, and followed by fever. It some- 
times subsided to recur again after a few days’ intermission. 
In certain instances there is a sudden chill, followed by high 
continuous fever from the onset. The period of rising tem- 
perature could not be ascertained with accuracy in most cases, 
because most patients were admitted to the hospital during the 
second week of the disease or later. 

Fastigium.—The stage of continued fever is not so prolonged 
nor the fever as high as is usually described for this stage. Out 
of 115 cases in which the temperature was studied carefully, 
only 8 showed a temperature over 40, and in only one case did 
the temperature reach 41. In Osler’s 1,118 cases, 67.48 per cent 
showed a temperature above 40, while in my series 43.4 showed 
a maximum temperature of 40 degrees. Osler quotes 25 per 
cent remaining below 40, whereas 44.34 per cent in the present 
series remained below this point. Another important deviation 
during this stage is the marked remission of temperature in the 
Manila cases, a condition which was noted in 24.34 per cent. 
This discussion is illustrated by Table III. 


TABLE III.—Temperature during the fastigium. 
Number of cases. Temperature. cates 
DT RE wh aes Bebe de onal 40° C. or below-------- 36. 52 
3. BRC SS = 3 SUS See es Se eee 88° toi 89-0? Foe ene 44,34 
LE See A Be eS or bod Trregular ...-..--=---- 9.56 


Stage of decline.—The fever in my series lasted on an average 
of thirty days. It began to show marked remissions and declines 


1X, B, 4 Gutierrez: Typhoid Fever in the Philippines 871 


at about the end of the third week. The temperature usually 
reached normal in about seven days, but in some cases the stage 
of decline was more prolonged and the temperature did not 
entirely subside until the eighth to the thirteenth day. How- 
ever, in no instances was the stage of decline as prolonged as 
the maximum given by Rogers. 

Recrudescence and relapse.—Manson in describing his expe- 
rience with typhoid in China says, ‘‘Not only is the tropical form 
grave from the outset, but extremely liable to relapse.” In the 
Manila series there were 6 cases of recrudescence, or 4.3 per 
cent, and 11 cases actually relapsed, or 8.02 per cent. In only 
one of these cases did the temperature relapse more than once. 
This patient is just recovering from the third attack at the time 
of writing. 

Irregular temperature.—Of the 115 cases in which the tem- 
perature was studied, in 11 cases, or 9.56 per cent, the temper- 
ature was exceedingly irregular. The fever presented many 
different phases, not one feature being constant. The temper- 
ature may be 39° or 40° for two or three days, and then quickly 
drop to 38° or 37°.5; or it may even drop to normal for two or 
three days, only to rise again to its former height. In one 
case the temperature was as low as 38° for a number of days 
and then became normal; this period in turn was followed by 
an intermittent type of 39° or more in the afternoons and 
normal in the morning. In this case, after thirteen days, the 
serum reaction was positive and culture of the urine showed 
typhoid bacilli at four different intervals after convalescence, 
finally becoming negative on the eighteenth day. 

Rigors.—Actual chills as described by Rogers were present in 
only 2.91 per cent of my cases. In two of these cases the rigors 
were caused by an intercurrent malaria, but the actual cause 
in the others could not be ascertained. 

Abortive cases.—There were 19 cases, or 13.86 per cent, in 
which the duration of the fever varied from five to fifteen days, 
or an average of eleven days from the onset, until the temperature 
became normal. During this period the temperature varied from 
38° to 39° C., rarely reaching the latter figure. The onset may 
be a typical steplike rise as is seen in classical cases of the disease. 
It may then become continuous for from two to five days, and 
then marked remissions appear and the temperature reach normal 
in a few days. These cases are interesting clinically as well as 
epidemiologically, because of obscurity of the diagnosis which 
rarely would be made outside of a hospital. Undoubtedly many 
such cases go through the course of the disease without being 

128443——5 


372 The Philippine Journal of Science 1914 


diagnosed as typhoid fever, especially as there are many fevers 
in the tropics of obscure etiology. These irregular types fre- 
quently are diagnosed as malaria or “‘fiebre continua.” 

In view of the above findings, there are a few diagnostic points 
that the clinician should constantly bear in mind when dealing 
with fever patients. In irregular types of typhoid the usual 
headache is generally present, sometimes intense but most fre- 
quently a dull ache is complained of at the top of the head. The 
relation of the fever to the pulse is maintained. A continued 
temperature of 39° C. with a pulse of 90 or 80 per minute is 
rather suspicious when noted in connection with the other above- 
mentioned symptoms. The spleen was enlarged at least to per- 
cussion in nearly all of these cases, and the typhoid tongue was 
present in all except the mildest cases. The serum reaction 
usually and blood culture almost invariably will, of course, con- 
firm the diagnosis. Therefore, in the tropics when we see a case 
of continuous or even intermittent fever with slow pulse, head- 
ache, and an enlarged spleen, we should be suspicious of its being 
typhoid, even though the fever is not high and the duration not 
as long as we expect to see in typhoid. The diagnosis in many 
of these cases is difficult or impossible except by laboratory 
methods. ‘Twelve of the cases in this series probably would not 
have been diagnosed outside of a hospital, where blood culture 
is routine in all doubtful fevers. 

Recently we have had a small epidemic of 12 cases of typhoid 
among the students in one of the Government dormitories in 
Manila. In all of these patients the disease was of short duration 
with an atypical fever, and other features of the disease were 
so irregular that the disease would not have been recognized 
except by blood cultures. 

The recognition of these atypical cases of typhoid is of partic- 
ular interest to public-health officials. An epidemic similar to 
the one mentioned above occurring in a country town, or other 
remote district, is liable to cause a great deal of trouble before 
the nature of the illness is recognized. This is particularly true 
in this country because the sanitary conditions are of the most 
primitive character. Frequently, there is no sewer system and 
the excreta is disposed of as the family sees fit. The usual 
method is to allow pigs and other scavengers to take care of it. 
Flies usually are abundant throughout the year, and the infective 
material is freely accessible to them. Clothes and utensils are 
never boiled, and the people around the patient probably eat with 
their fingers without so much as washing their hands with soap 
and water. 


IX, B, 4 Gutierrez: Typhoid Fever in the Philippines 373 


ANALYSIS OF SYMPTOMS 


General appearance.—In frank cases, except in the mildest, 
the typhoid clinical picture is present. “Flushed faces” were 
noted in 51 cases, typhoid tongue was noted in 108, typhoid facies 
in 81, typhoid state in 25, bleeding from the gums and lips 
was found in but 8 cases, and “rose spots” were found in 26 
cases, although they were not especially looked for. It has 
been a common belief among practitioners about Manila that 
rose spots are rare in typhoid among Filipinos. I have not been 
able to find any evidence to justify this conclusion. If rose 
spots are not especially searched for, they will be overlooked 
oftener in the dark-skinned race than will be the case in fair- 
skinned patients. In the last 15 cases of this series, rose spots 
were searched for with care, with the result that they were 
found in 11, or 73.66 per cent, of the patients. 

Delirium probably is not as prevalent as it is in temperate 
climates. It was present in 19 of my cases, or 13.86 per cent. 
Seven cases were of the low-muttering type, and 12 patients 
had wild delirium. The lowered incidence of delirium is prob- 
ably due to the unusually low fever encountered in many of our 
patients. “Typhoid state’ was present in only 18.24 per cent 
of this series. 

The circulatory system.—tThe heart presented the usual toxic 
effect of this disease, and no peculiar conditions were noted. 
The usual pulse found in typhoid fever was encountered in this 
series. The rate was disproportionately low as compared with 
the fever, and was one of the important symptoms that was 
looked for, especially when the fever was irregular. 


TABLE 1V.—Pulse rate in typhoid. 


| ‘throughout high | for twoor more. | Pulse not over 100 

temperature. days. : 

Number.| Per cent.| Number. |Per cent. | Number.| Per cent. 

10 UE Se pa en een rr ee a 47 55. 95 24 28.57 13 15.47 

Sie CE eae ae ae ee eee See 16 35. 71 10 35.70 8 28.35 

EIGEN sen oe ee ee ee on oie ae (ES es 3 23.07 10 76, 92 
Rogers: 

PT) Beer ore oe Seen ee ee ee 22 36. 06 26 33.30 18 30. 00 

Mera al eis 8 sa ees Sr eee | ee OE ee i, 11.10 16 88. 90 

Childrents. - 2st) wre seen. Su eee eee SS Se |---------- jesseeesone 27 | 100.00 

I 


The spleen.—Probably next in importance to the pulse rate 
comes the spleen. Of 118 cases, the spleen index was recorded 
in 31. In 16 it was enlarged 2 centimeters or more below the 


374 The Philippine Journal of Science 1914 


arch; in 11 cases it was enlarged to percussion; in 4 cases it 
was not enlarged at all; and in 14 cases it was not mentioned. 
Only one case showed an enlargement of 10 centimeters below 
the costal arch. The diagnostic significance of enlargement of 
the spleen is greatly reduced in the tropics, on account of the 
many diseases giving rise to an enlargement of this organ. 
However, when viewed in connection with the other symptoms, 
it has some value in this respect. 

The respiratory system.—The lungs showed the usual com- 
plications encountered in this disease. Bronchitis was found in 
55 cases. Pneumonic consolidation was noted in 13 cases and 
tuberculosis in 6 cases. In two of the tubercular patients a 
latent infection was probably excited by the typhoid, because 
the patients showed no active lesions of tuberculosis at the time 
of entrance. 

The digestive system—The typhoid tongue was fairly con- 
stant, being present in 108, or 78.83 per cent. Tympanitis has 
not been so marked as to cause alarm; in only one was it a 
very severe symptom. The bowels were more often constipated 
than loose. There were 9 cases that showed diarrhcea through- 
out the disease, making 6.56 per cent of the cases, and 21 
cases, or 15.36 per cent, had transient diarrhea. The rest 
were constipated throughout the course of the illness. 

Intestinal hemorrhage—tThis complication occurred in 21 
cases, or 15.23 per cent of the series. In one patient with hem- 
orrhage, diarrhcea persisted throughout the course of the 
disease, while two other cases showed transient diarrhoea, thus 
making a total of 3 cases which showed diarrhea, or 13.33 per 
cent of patients with hemorrhage associated with diarrhoea in 
the same person. In 8 cases with hemorrhage, or 38.14 per 
cent, special attention was called to the severity of the disease. 
Out of the 21 cases with hemorrhage, 11 lived and 10 termi- 
nated fatally. 

Intestinal perforation.—Probably next in frequency to hzemor- 
rhage is perforation. In this series there were 10 patients 
who had perforation, or 7.29 per cent of the total cases. This 
figure probably is higher than the normal percentage of per- 
forations in the Philippines, because frequently only the most 
severe cases, or those with complications, are sent to the hospital. 
Three cases in this series had general peritonitis when they 
were admitted. 

Malarial incidence.—Intermittent malarial infection was found 
four times in this series. In one, the fever was ushered in 
with chills and the malaria was not discovered until its re- 


IX, B, 4 Gutierrez: Typhoid Fever in the Philippines 875 


appearance during convalescence from the typhoid. In one of 
the four cases malarial symptoms predominated. In two cases 
the stage of fastigium was somewhat irregular, the temperature 
varying greatly. Repeated doses of quinine, both intravenously 
and by mouth, made the temperature more regular, but other- 
wise failed to modify the course of the disease. The double 
infection was verified by blood examination in each case. 

Other complications.—Periostitis was found in 5 cases, throm- 
bosis of the left femoral in 1, and typhoid spine in 1. Post- 
typhoid psychosis or mania was encountered three times, but 
all of the patients affected recovered. Peripheral neuritis was 
encountered in a number of cases, but the total number could 
not be ascertained as they were not recorded in every case. 

Mortality —Osler gives 7.5 per cent as the mortality rate 
in his Baltimore series. However, he states that the death 
rate varies from 5 to 12 in private practice and from 7.03 to 
12 per cent in hospital practice. Curschmann places it at 12.7 
per cent, and Rogers at 16.3, for white troops in the tropics 
and at 26 per cent for the Indians. When we come to the 
Philippines we find Chamberlain placing it at 17.65 per cent for 
the Filipinos, while 16.8 per cent was the figure set for Americans. 
In this series there were 20 deaths, or about 20.43 per cent. 
Upon closer examination it was found that about half of these 
cases were admitted in a dying condition, or, if not actually 
dying, they were in a hopeless condition. ‘There were as follows: 
three cases with general peritonitis; 4 patients lived only two or 
three days after admission; 3 were admitted from the twenty- 
third to the sixtieth day of the disease; and another 3 cases 
came in after the fifteenth day of the disease. Excluding 10 
cases which certainly were hopeless from the time they came 
in, we have a mortality of 13.13 per cent. This is certainly 
a high percentage of mortality, but it compares favorably with 
those cases of Rogers, Chamberlain, and Curschmann. Outside 
of the delayed hospitalization, a serious handicap is the generally 
poorly nourished condition among most of the Filipinos. The 
class of patients dealt with in this article are mostly from the 
poorer class, many of whom cannot afford the services of an 
outside physician, so that alimentation and intestinal disorders 
are wholly neglected. 

Laboratory method of diagnosis.—There does not appear to 
be any unusual disturbance of the blood picture. From a 
diagnostic standpoint the leucocyte count is interesting, and it 
is especially so in the tropics because of the increased difficulties 
of diagnosis. 


376 The Philippine Journal of Science 1914 


The number of leucocytes in the various cases in this series 
was as follows: 
TABLE V.—Cases with leucocytes. 


Leucocytes. Cases. 
From 2,000 to 3,000 3 
3,000 4,000 12 

4,000 5,000 19 

5,000 6,000 27 

6,000 7,000 18 

7,000 8,000 16 

8,000 9,000 8 

9,000 10,000 9 

10,000 13,000 7 

Of twenty differential counts made, there was a decrease to 
59.8 per cent of the polymorphonuclears, while in about 50 per 
cent of these cases the lymphocytes exceeded 30 per cent. 
Large mononuclears were 3.7; transitionals, 4 per cent; and 
eosinophiles, 1.65 per cent. The hemoglobin estimation had an 
average of 84 per cent by the Tallqvist method. 

Serum reactions were made 247 times, the dilutions being 
1-40, the time one hour. Olser considers this an almost specific 
diagnostic method for typhoid; certainly for routine methods 
it serves the purpose. During the fiscal year 1912, laboratory 
facilities were limited and frequently only one reaction was 
obtained. During that year 37 out of 47, or 78.72 per cent 
of the suspected cases, gave a positive reaction. During the 
fiscal year 1913, the agglutination test was made more frequently 
and several tests were often made on the same patient, par- 
ticularly when a negative report was received with suggestive 
clinical findings. 

TABLE VI.—Serum reaction. 


Negative, zs. | Positive, zy. Positive, zo. 
| | 


= . 
vo 

i i : oO Lo] x] ] 
Period of disease. : 3 3 3 3 g 3 ® ® Z 
w 3 o ° on) o ° i) oO ts} as | 3 i) 
g/ S/o (3/8) 5) 81/8) 8) 8\8) 8] & 
»lalelelealeal|elala|el/e|ea Aa 
eb Te go 1912) | neues | ee iy Wee 2 ha ek | i Ha 2| 1] 50.00 
1913] 8] 7 10)|) A 899) Bil) 2) 2719) "e7s00 
sSesdosiid sek sei | TESTES Sia GH Pe ea SE ae Ol Nae 30 | 23 | 76.66 
1918| 5) 8| 2] 58) 4) 4] GO] 4] 4/ 108) i108 | 95.87 
igi he | TTD, Ne PAN eae ee 10 bse Pilea ee 15 | 13 | 86.66 
O13) eae dl sees 32| 2| 2] 32| 2] 2] 65| 64] 98.48 
Total | 19128 MeaON Welln 4. (kop |e ibe eae 47 | 37 | 78.72 
ta j9is| 14| 11! 9] 95] 10/ 9] 91] 9] 8] 200} 186 | 93.00 
Grand total __-_-_-|_----- 24| 17] 18 | 120} 10! 9{|103| 9| 8| 247 | 228 | 90.28 
Rogers’ 2 ee 5 ee 31 | 22 | 20] 10 1/ 1] 24| 2| 2| 150 | 119 | 79.80 


IX, B, 4 Gutierrez: Typhoid Fever in the Philippines 3877 


Blood cultures were not taken in all of these cases, owing to 
the patients coming to the hospital late in the course of the dis- 
ease. However, 45 blood cultures were taken by Dr. HE. H. 
Ruediger of the Bureau of Science, and of the 13 cases in which 
the blood was taken during the first six days of the disease 8 
cases, or 61.53 per cent, were positive for Bacillus typhosus. 
After the sixth day of the disease there were 32 blood cultures 
taken with 12 positive findings, or 37.5 per cent. 


SUMMARY 


1. The age incidence of typhoid fever as seen in the Philip- 
pine General Hospital corresponds with that of other countries, 
but is not so high as claimed by Rogers, Manson, and Nichols. 

2. The fever course of the disease is essentially the same as 
in the United States and in Germany, but the temperature is 
much lower and the stage of fastigium is not as typical. 

3. Relapses occur about as frequently as in other countries. 

4. Abortive cases are rather common and often difficult to 
diagnose. Every continued fever with slow pulse and enlarged 
spleen should have a serum reaction and other confirmatory 
- tests. Blood culture should be employed in all cases of doubt. 

5. Delirium and the typhoid state are not so common as in 
other countries. This is probably due to the lower temperature 
in typhoid in the Philippines. 

6. Intestinal hemorrhage occurred in 15.23 per cent of all 
cases. I am unable to ascertain the cause of such a large num- 
ber. It may be due to neglect of the gastrointestinal canal con- 
sequent to delay in receiving medical attention. 

7. Perforation is also common, for which no special cause 
can be attributed. 

8. The mortality on superficial examination seems to be high, 
but after eliminating these cases that came to the hospital in a 
dying or hopeless condition we obtain a more normal death rate. 
The further slightly higher percentage of death rate may be 
due to the lower resistance of the Filipinos consequent to their 
modes of life, undevelopment, and limited diet. 

Since this paper was written, the number of intestinal hemor- 
rhages and mortality has been decreased, because the cases were 
admitted to the hospital earlier in the course of the disease and 
the diet of the people in general has been improved. 

In conclusion I wish to express my sincere gratitude to Dr. 
W. E. Musgrave for his interest in the course of the preparation 
of this article and to Dr. L. Gomez for his suggestions. 


378 The Philippine Journal of Science 


REFERENCES 


BRUNS, EARL H. Milit. Surgeon (1908), 22, 369. 

CASTELLANI, A., and CHatMeErRS, A. J. Tropical Medicine. Wm. Woodcock 
& Co., N. Y. (1910). 

CHAMBERLAIN, W. P. Phil. Journ. Sci., Sec. B (1911), 6, 299. 

CHAMBERLAIN, W. P., and BLOOMBERGH, H. D. Miulit. Surgeon (1910), 27, 
529. 

Editorial. Ibid (1910), 26, 586. 

EDWARDS, ARTHUR R. Principles and Practice of Medicine. Lea Febriger, 
N. ¥. (1909); 

GooDHUE, E. S. Am. Journ. Pub. Hlth. (1912) 2, 883-895. 

HEISEeR, V.G. Phil. Journ. Sct., Sec. B (1910), 5, 171. 

JACKSON, T. W. Med. Rec. (1912), 76, 511. 

LA Garpe, L. A. Milit. Surgeon (1908), 22, 373. 

MANSON, P. Tropical Diseases. Wm. Woodcock & Co., New York (1911). 

NicHous, H. J. Milit. Surgeon (1909), 22, 365. 

OsLER’S Modern Medicine. Lea Brothers & Co., Philadelphia and New York 
(1907), 2. 

PHALEN, J. H., and KILBOURNE, E. D. Milit. Surgeon (1910), 27, 56. 

Rocers, L. Fevers in the tropics (1910), 110-148. 


REVIEWS 


Biochemic Drug Assay | Methods | with special reference to the phar- | maco- 
dynamic standardization of drugs | by | Paul S. Pittenger, Ph. G., 
Ph. C., Phar. D. | instructor [etc. 5 lines] | edited by F. E. Stewart, 
M. D., Ph. G. | professor [etc. 7 lines] | Philadelphia | P. Blakiston’s 
Son & Co. | 1012 Walnut Street | Cloth, pp. i-xiv + 1-158. 89 illustra- 
tions and diagrams. Price $1.50 net. 

The book is not exactly what the title—_A Manual of Biochemic 
Drug Assay Methods—would lead one to believe as the author 
has included only those methods of drug assaying which are 
physiological in nature. Although it represents the first attempt 
at preparing a manual of this sort, it contains very little that is 
original in material or in the arrangement thereof. 

The contents of the book are grouped into eight chapters; 
chapter I is headed Preliminary Considerations and deals pri- 
marily with the factors making drug assaying desirable; chap- 
ters II to VI, inclusive, are devoted to descriptions of the more 
important physiological assay methods and their application to 
the standardization of the cardiac stimulants and depressants, 
epinephrine and products of the suprarenal gland, ergot, pitui- 
tary extracts, and cannabis indica; chapter VII contains descrip- 
tions of apparatus and technique employed in conducting the 
assays described in previous chapters; and chapter VIII gives 
directions for preparing a few of the stock solutions commonly 
used in work of this nature. 

As stated in the preface, the manual is intended for students 
of pharmacy, students of medicine, and experts engaged in drug 
standardization work. With respect to students in pharmacy, 
the author presupposes such students to have a very considerable 
knowledge of animal anatomy and practical physiology. Until 
such studies are made a part of the curriculum of the schools 
or colleges of pharmacy, the manual will be beyond the ordinary 
student of pharmacy. To the student of medicine who is accus- 
tomed to the use of a textbook on pharmacology, such as that of 
Cushny or of Sollman, the manual will seem elementary indeed 
and will be found of little value. It should, however, be of some 
service to experts engaged in the standardization of drugs or to 
those who desire a general knowledge of the physiological 
methods employed in the standardization of drugs without the 
trouble of consulting the larger works on pharmacology or of 
looking up the original references. 

A. G. DUMEZz. 
379 


380 The Philippine Journal of Science 


Beriberi | by | Edward B. Vedder, A. M., M. D. | Captain Medical Corps, 
U. S. Army. Member of the United States Army Board | for the 
Study of Tropical Diseases as They Exist in the Philippines, | Decem- 
ber, 1910, to April, 1913. | [dash] | The Cartwright Prize of the 
Alumni of the College of Physicians and Surgeons, | Medical Depart- 
ment of Columbia University, New York for 1918, and | published by 
permission of the Surgeon-General of U. S. Army. | [dash] | Illus- 
trated by Numerous Engravings | and by Five Colored Plates | [dash] 
| New York | William Wood and Company | MDCCCXIII. Cloth, 
pp. i-viii + 1-427. 

This book is the most extensive treatise on beriberi in the 

English language, and appears at a time when not only all 

medical scientists but also the lawmakers in several countries are 

interested in the subject. Vedder has had very favorable op- 
portunity for carrying on investigations of the etiology of beri- 
beri and its cure. As a member of the United States Army 

Board for the Study of Tropical Diseases as They Exist in the 

Philippines, his experience in Manila made it possible for him 

_to seeemany cases of the disease and to prove the value of the 
extract of rice polishings prepared by Chamberlain and Vedder. 

The book deals with the history and the clinical and pathological 

aspects of the disease, especial emphasis being placed on the 

etiology. His conclusions that avian polyneuritis is the same 
disease as human beriberi and that beriberi is caused by a too 
exclusive diet of polished rice or by any other diet lacking in 
the vitamines which are necessary for normal metabolism are 
now accepted by the majority of workers on the subject, and 

Vedder’s review of the work of himself and others is presented 

in a very pleasant and convincing manner. The abundant illus- 

trations are excellent, and the book is presented in Wm. Wood 

& Co.’s usual good form. B. C. C. 


The Mechanistic Principle and | the Non-Mechanical | an inquiry into funda- 
mentals with extracts from | representatives of either side | by | 
Paul Carus | Chicago | The Open Court Publishing Company | 1913 | 
Cloth, pp. i-iv + 1-125. 


Memory | lectures on the specific energies | of the nervous system | by | 
Prof. Ewald Hering | University of Leipzig | fourth edition, enlarged | 
1913 | The Open Court Publishing Company | Chicago London | Cloth, 
pp. 1-70. 


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MALARIA IN THE PHILIPPINE ISLANDS 


I. EXPERIMENTS ON THE TRANSMISSION OF MALARIA WITH ANOPHELES 
(MYZOMYIA) FEBRIFER SP. NOV., ANOPHELES (PSEUDOMYZOMYIA) 
ROSSII, ANOPHELES (MYZORHYNCHUS) BARBIROSTRIS, ANOPH- 

ELES (MYZORHYNCHUS) SINENSIS, AND ANOPHELES 
(NYSSORHYNCHUS) MACULATUS * 


By ERNEST LINWOOD WALKER and MARSHALL A. BARBER 
(From the Biological Laboratory, Bureau of Science, Manila, P. I.) 


I. INTRODUCTION 


The efficient and economical conduction of an antimalarial 
campaign in any country must be based not only upon accurate 
information of the anopheline mosquitoes and their distribution, 
but also upon the experimental determination of the ability of 
the different species to transmit malaria. As a practical exam- 
ple of the importance of this knowledge, it has been estimated * 
that in the sanitation of the Panama Canal Zone between 100,000 
and 250,000 dollars were saved by the knowledge that a certain 
anopheline mosquito, Anopheles malefactor, which breeds in the 
collections of water in hollow stumps, resulting from the ex- 
tensive deforestation of the country, was unable to transmit 
malaria. 

Of about 100 species of Anophelinz now known, probably less 
than one-third have been definitely proved, either by the dis- 


* Part II of this work, The distribution of the more common anophelines 
and the distribution of malaria, will appear in an early number of this 
Journal. , 

*Carter, H. R., Am. Journ. Trop. Dis. & Prevent. Med. (1913) 1, 48. 

129558 381 


382 The Philippine Journal of Science 1914 


section of the naturally infected mosquitoes or by experimental 
infections, to be capable of transmitting malaria. The remaining 
species either have not been investigated or the evidence of their 
susceptibility is inconclusive or contradictory. Furthermore, it 
has been demonstrated that among the anophelines capable of 
transmitting malaria the different species vary widely in their 
susceptibility to infection with the malarial parasite and that 
the same species may not be capable of serving as vector of all 
three types of human malaria. Finally, it should not be over- 
looked that the same species of anopheline may vary in its sus- 
ceptibility to infection with the malarial parasite in different 
countries or under different ecologic conditions. Such variations 
have been recorded in the susceptibility of Glossina morsitans 
to infection with Typanosoma rhodesiense. A similar variation 
in susceptibility may account in part for the contradictory results 
obtained by different authors in their dissections of, or exper- 
iments with, certain anopheline mosquitoes. 

These facts and possibilities make it impossible to predict the 
susceptibility of any species of Anopheline to infection with the 
malarial parasite, and render it imperative that every species 
be tested experimentally in order to be certain of the réle played 
by it in the dissemination of malaria in any country. 

There are two methods of determining the susceptibility of an 
anopheline mosquito to infection with the malarial parasites. 
One of these is the dissection of anophelines caught in houses 
harboring malarial patients, in order to determine the presence 
of odcysts or sporozoites of the parasites in the mosquitoes: the 
other consists in experimentally infecting mosquitoes bred from 
larve, and afterwards dissecting the mosquitoes for the discovery 
of the developing parasites, or allowing them to bite healthy 
persons to prove their ability to transmit the infection. 

It is frequently stated in the literature that such a mosquito 
has been experimentally infected with malarial parasites but 
has not been found infected in nature, with the implied conclu- 
sion that experimental infections furnish a less reliable index 
of the ability of the species to transmit malaria. However, we 
believe that the experimental infections, if properly conducted, 
supply the more reliable index of the relative susceptibility of 
different species. In mosquitoes examined for natural infection 
with malarial parasites it is impossible to determine whether or 
not the mosquito has had an opportunity to bite an infected 
person, whether or not if the opportunity presented itself it 


* Kinghorn and York, Ann. Trop. Med. & Parasit. (1912), 6, 269-285. 


1x,B,5 Walker and Barber: Malaria in the Philippines 383 


was taken advantage of, if the mosquito did suck blood whether 
or not the blood contained gametes sufficiently numerous to 
- infect the mosquito, and whether or not sufficient time had 
elapsed after the feeding for the parasites to attain develop- 
ment. In properly conducted experimental infections all of 
these essential factors are under complete control. We are 
also of the opinion that it is strong, if not wholly sufficient, 
proof of the capacity of a species of mosquito to transmit mala- 
ria if it can be infected with the malarial parasite and if the 
parasites develop sporozoites and infect the salivary glands, 
without subjecting healthy persons to the bites of the experimen- 
tally infected mosquitoes. It is improbable, although possible, 
that sporozoites should be developed, migrate to and infect the 
salivary gland of the mosquito, and yet be incapable of being 
injected into or of infecting man. As it is known that different 
species vary greatly in their ability to transmit malaria, it is 
of the greatest practical importance to determine the relative 
susceptibility of the different anophelines in any country. In 
most infection experiments hitherto performed, careful com- 
parative tests have not been made to determine the relative 
susceptibility of the different species of anophelines. In some 
cases a rough estimation of the proportion of a given species 
that becomes infected has been obtained. However, to be of 
practical value, the test should be comparative of different spe- 
cies of anophelines in a given region. This can be accurately 
determined only by a large series of experiments in which the 
different species are fed at the same time upon the same mala- 
ria patients and in which only those mosquitoes that sucked a 
full meal of the infected blood are selected for dissection. 
Under these conditions of experimentation the percentages of 
infected mosquitoes, and the relative number of odcysts in the 
infected individual of the several species, will give a reliable 
index of the relative susceptibility of these anophelines to 
infection with the malarial parasite. 

Susceptibility, while the most important, is not the only factor 
in determining the importance of any species of mosquito in the 
transmission of malaria. Geographical distribution and preva- 
lence of the species, its habitat in relation to the dwellings of 
man, and its avidity for human blood also play a more or less 
important part in determining the role of the different species. 
All of these factors must be determined for each species before 
we can estimate accurately their relative importance in the 
epidemiology of malaria in any country. 

In the Philippine Islands the species and distribution of the 


384 The Philippine Journal of Science 1914 


anopheline mosquitoes have been extensively, but not intensively 
and exhaustively, studied by Giles (1904), Banks (1906), Lud- 
low (1908), and others. The following species have been - 
credited to the Philippine Islands :* 


Myzomyia funesta Giles. Pyretophorus pitchfordi Giles. 
Myzomyia thornton Ludlow. Myzorhynchus barbirostris van 
Myzomyia mangyana Banks. de Wulp. 
Pseudomyzomyia rossi Giles. Myzorhynchus pseudobarbirostris 
Pseudomyzomyia ludlowii Theo- Ludlow. 

bald. Myzorhynchus sinensis Wiede- 
Psuedomyzomyia indefinata Lud- mann. $ 

low. Myzorhynchus vanus Walker. — 
Stethomyia pallida Ludlow. Nyssorhynchus fuliginosus Giles. 
Pyretophorus minimus Theobald. Nyssorhynchus philippinensis Lud- 
Pyretophorus freere Banks. low. 


Pyretophorus philippinensis Lud- Celia kochi Dénitz. 
low. 

It is probable that a revision of the synonymy of these species 
would reduce the number as well as change the names of some 
of these species, and it is certain that a more intensive study 
of the anopheline fauna of these Islands would disclose other 
species. As they stand, only 6 of the 17 species are reputed to 
be capable of transmitting malaria. Of these, Anopheles funesta 
probably does not occur in the Philippines, the capability of 
Anopheles rossti to transmit malaria is disputed, and the specific 
validity of Anopheles ludlowii is doubtful. 

Of the 5 species considered in this paper, namely, Anopheles 
febrifer Banks sp. nov., Anopheles rossii Giles, Anopheles bar- 
birostris van de Wulp, Anopheles sinensis Wiedemann, and 
Anopheles maculatus Theobald emend. Stanton, 2 are not con- 
tained in the above list and have not hitherto been credited to 
the Philippine Islands and 1 is a new species. 

Anopheles febrifer was found very prevalent in Laguna 
Province. We have been unable to identify it with any of the 
hitherto described species of anophelines, although for a time 
we were inclined to believe it was the same as that identified by 
Ludlow in the Philippines as Myzomyia funesta. Mr. Banks, en- 
tomologist of the Bureau of Science, to whom it was referred, 
decided that it was a new species and named it Myzomyia febri- 
fer... The more distinctive characters of the female of this 
species are as follows: 

A small anopheline mosquito. Head dark brown, with white 
upright forked scales in the middle and a tuft of white hairs 


* Banks (1906). 
*This Journal, Sec. D (1914), 8, No. 4. See also the note on page 439. 


Ix,B,5 Walker and Barber: Malaria in the Philippines 3885 


projecting forward. Palpi dark brown, with two broad cream- 
white apical bands, one of which is terminal, separated by a 
narrow dark band, and one narrow white basal band. Probos- 
cis dark brown with a tawny tip. Thorax light brown on the 
back and dark brown on the sides, and covered with hairlike 
curved scales. Wings spotted, with the light color slightly pre- 
dominating; the front edge black, with 5 pale yellow spots and 
a pale-yellow tip; the fringe with light spots at the end of all 
of the veins except the sixth. Legs dark brown, with very 
narrow light bands at the joints. Abdomen dark brown, hairy. 

The larve of Anopheles febrifer were found very abundant 
in shaded brooks of Laguna Province. They are found in 
depressions of the bank or under overhanging portions of the 
bank in the wooded streams, and especially where collections 
of drift twigs and leaves or tufts of rootlets were projecting 
into the water. This species also breeds in open brooks or 
irrigation ditches, if they contain running water and there is 
overhanging grass or other vegetation to furnish shade. As this 
is a hitherto unknown species, there are no data on its suscepti- 
bility to infection with, and its ability to transmit, malaria. 

Anopheles rossii was first described by Giles in 1899. It is 
credited to the Philippines by Banks (1906) and Ludlow (1908). 
The synonymy of this species is as follows: 

Anopheles rossii Giles, 1899. 

Anopheles vagus Dénitz, 1902. 

Myzomyia rossii Theobald, 1903. 
Pseudomyzomyia rossii Theobald, 1907. 
Nyssomyzomyia rossii James and Liston, 1911. 

Anopheles ludlowii is a species nearly related to, if not a 
variety of, Anopheles rossii. It was first discovered by Ludlow 
in the Philippine Islands in 19038. She considered it to be a 
new species and sent it to Theobald, who named it. The syn- 
onymy of this species is as follows: 

Anopheles ludlowii Theobald, 1903. 
Myzomyia ludlowii Theobald, 1903. 
Nyssomyzomyia ludlowii James and Liston, 1911. 

The differential characters separating ludlowii from rossi 
are given by Theobald (1903) as follows: 

A very variable species, somewhat like rossi at first sight, but easily 
told by the spotted (yellow) legs and much shorter fork-cells. The base 
of the first submarginal cell is always slightly nearer the apex of the 
wing, and the costal spots differ slightly, but are to some extent variable. 


The cross veins are most unstable. The palpi are very similar, but the 
apical band in rossii is rather longer. The chief difference is that in 


386 The Philippine Journal of Science 1914 


rossti the second white band is a third of the way down the palpi; in this 
species (ludlowti) it is less, and the black intervening area is much smaller. 

James and Stanton (1912) list the two as distinct species. 

Stanton (1913) distinguishes the two species, but states that 
Anopheles ludloww is nearly allied to Anopheles rossit. 

Alcock (1913) describes the two species, but says that if it 
were not for the fact that Anopheles ludlowii is said to transmit 
malaria in the Andamans he would consider it a variety of 
Anopheles rossii. 

Knab (1913) considers Anopheles ludlowii as a species closely 
resembling, but distinct from, Anopheles rossi. 

Mr. Banks, entomologist of the Bureau of Science, considers 
rosst and ludlowii as variations in one and the same species, 
namely, Anopheles rossii. He states that this opinion is based 
upon examination of the type specimens of these two forms at 
the British Museum several years ago and that Theobald agreed 
with him at that time that lwdlowti was not a distinct species. 

All of the mosquitoes of the rossii-ludlowii group collected by 
us, both from fresh water in Laguna Province and from brackish 
water along Manila Bay in Rizal Province, have been of the 
rossti type. Since these experiments were completed, one of 
us (Barber) has collected in fresh water in Mindoro larve from 
which were bred an Anopheles with conspicuously spotted legs, 
possibly ludlowii. Our experimental infections reported in this 
paper have all been made with Anopheles rossii. ~ 

Anopheles rossii is, according to the observation of Banks 
(1906), Ludlow (1908), and ourselves, one of the most, if not 
the most, prevalent and widely distributed anophelines in the 
Philippines. This species may be found breeding everywhere 
in water open to the sunlight, both in fresh water and in the 
brackish or salt water of esteros along the extensive coast of 
the Archipelago. It will even breed in stagnant water if run- 
ning water is not available. We have found the larve of this 
species in the esteros along Manila Bay, in salt beds used by the 
natives for evaporating sea water, in irrigated rice fields, open 
rivers, the overflow from artesian wells, carabao tracks filled 
with water, about water holes and troughs, carabao wallows, 
and even in the foul water of tanks used to soak sugar-cane 
stalks. 

The evidence of the ability of Anopheles rossti and Anopheles 
ludlowii to transmit malaria is exceedingly contradictory. 

In India all investigators appear to be agreed that Anopheles 
rossii, although one of the most prevalent species, is not of 


ix, B,5 Walker and Barber: Malaria in the Philippines 387 


practical importance in the transmission of malaria. Liihe 
(1906) gives the following table of comparative results of 
dissections by different authors of Anopheles culicifacies and 
Anopheles rossii captured in malarial houses in India: 


TABLE I.— Malaria parasites found in Anopheles culicifacies and Anopheles 
rossi caught in houses in India. 


Anopheles (Myzo- | Anopheles (Myzo- 
myia) culicifacies. myia) rossit. 


Place. Observer. RSithiones Leal 
: rozoites . rozoites 
sected ao sected. ce 
glands. glands. 
Per cent. 
Mise Mivnn= soon oe oe Stephens and Christophers-_|-_---.-----|---------- 324 0 
1B Ne tae Caer coon Bee pe James s220 ee eR ed 259 12-4.6 496 0 
Ennur (fishing village near | Stephens and Christophers. 69 6-8.6 364 0 
Madras).® | 
LOL dee 2 ei ie ghee Sl MINES coe ne ae see ee ea aca me eae sierra ane 18 0 
Near Madrase ___.__.___-__ Cornwallhe eee 25 | 4-16 35 0 


% Anopheles rossii very rarely in the houses. Anopheles culicifacies incomparably less 
frequent ; the 69 specimens are the total collection for almost a week. 

>In one of these 18 specimens, however, odcysts were found. 

© Both species of mosquito were collected in the same houses and under the same conditions. 


James (1902), however, states that he has obtained positive 
results in experimental infections of Anopheles rossti with sim- 
ple tertian, malignant tertian, and quartan malarial parasites. 

Schiiffner (1902) worked on the experimental transmission 
of malaria at Deli, Sumatra, with a mosquito which he designates 
as “Anopheles I.” From his description and especially from 
his figures of this mosquito it is very probable that it was Ano- 
pheles rossii. Eysell (1910) is also of this opinion. Schiiffner 
obtained undoubted infections of this species of mosquito with 
both tertian and subtertian malaria, as the excellent figures of 
sections of the mid-gut and salivary glands in his plates demon- 
strate, and he was successful in transmitting the infection with 
both types of malaria to healthy persons by bites of the experi- 
mentally infected mosquitoes. 

Banks (1907) was the first investigator to determine experi- 
mentally the ability of identified specimens of Anopheles ludlowii 
to transmit malaria. He succeeded in infecting, and securing 
development of the malarial parasites up to the sporozoite stage 
in, mosquitoes which he found as larve in brackish water at Olon- 
gapo, Luzon, P. I., and which he identified as Myzomyia ludlowii 
by feeding them on the blood of a patient infected with subter- 


388 The Philippine Journal of Science 1914 


tian malarial parasites. He was furthermore able to transmit 
the disease by allowing the experimentally infected mosquito to 
bite a healthy man who had volunteered to submit to the experi- 
ment. Banks now considers the Myzomyia ludlowi with which 
he experimented to be identical with Anopheles (Myzomyia) 
rossit. 

In 1910 De Vogel infected and secured development of the 
malarial parasites up to the odcyst stage in mosquitoes bred 
from the larve collected at Samarang, Java. He failed to follow 
the development of the parasites in the mosquitoes further 
because of the difficulties in keeping the mosquitoes alive suf- 
ficiently long in captivity. Infections were obtained with indi- 
viduals bred from larve collected from brackish or salt water 
and not with individuals of the same species bred from the larvze 
obtained from fresh water. De Vogel first identified the mos- 
quitoes breeding in brackish or salt water as Anopheles vagus 
D6nitz, which is considered by some as a synonym of Anopheles 
rossii Giles. Specimens were submitted to Professor De Meyero 
of Amsterdam, and he declared them to be Myzomyia rossii. 
Individuals hatched from the same lot of larvee as those infected 
were submitted to Theobald, who likewise fost ee them to 
be Myzomyia rossii. 

Strong (1910), in the discussion of De Vogel’s paper at the 
first biennial congress of the Far Eastern Association of Trop- 
ical Medicine held at Manila in 1910, made the following 
statement with reference to the transmission of malaria by 
Anopheles ludlowii or rossii in the Philippine Islands: 

During the past year in connection with the work in the courses of trop- 
ical medicine in the Philippine Medical School relating to the study of 
malaria, we attempted to infect numerous specimens of Myzomyia rossi by 
exposing patients suffering with severe cases of zstivo-autumnal and tertian 
malaria to their bites. However, although these experiments were extensive 
and were carried on over a period of several months during the autumn, 
they were entirely unsuccessful. In no case did the dissection of any of 
these mosquitoes, although a large number were examined, reveal any 
odcysts in the walls of the stomach, and in the study of stained sections made 
of the salivary glands no sporozoites could be detected. Later attempts to 
infect other human beings by the bites of specimens of Myzomyia rossii, 
which had been previously fed on the blood of patients suffering with severe 
malaria and whose blood certainly contained gametes, also failed. The 
larve of these mosquitoes were collected in the estuaries about the city. 

Strong states that it is now known that Myzomyia ludlowii 
and M. rossii are one and the same species. 

Christophers (1912) found Anopheles (Nyssomyzomyia) lud- 
lowii, a species which breeds in and about salt swamps and which 
is not found at a greater distance than a kilometer and a half 


pl i 


~~ 


; 


1x,B,5 Walker and Barber: Malaria in the Philippines 889 


from salt or brackish water, to be the chief carrier of malaria 
in the Andamans. The author is doubtful whether any part is 
taken in the transmission of malaria here by the other common 
species, Nyssomyzomyia rossii and Myzorhynchus barbirostris. 

Stanton (1913) states that a large series of dissections and 
infection experiments, carried on by him in the Federated Malay 
States, failed to show any development of malarial parasites in 
Anopheles rossit. 

Anopheles barbirostris is a common Malayan species, having 
the following synonymy: 

Anopheles barbirostris van de Wulp, 1884. 
Myzorhynchus barbirostris Theobald, 1908. 

This species is reported in the Philippines by Banks (1906) 
and Ludlow (1908). We have found it breeding scatteringly 
but widely spread in Laguna Province. It was first found 
breeding in December in a semistagnant pool, which was densely 
shaded and contained growths of duck week, pond lilies, and alge. 
Later it was found breeding in open rivers and brooks, asso- 
ciated with Anopheles rossii, and to a lesser extent in shaded 
brooks, associated with Anopheles febrifer. Larve of this 
species are particularly to be found in collections of driftwood 
and dead leaves at the lower end of the semistagnant pools and 
under overhanging vegetation along the banks of open streams. 
The larve are rarely numerous and are usually found only 
scatteringly. 

James (1902) states that he was successful in infecting and 
securing development of odcysts and sporozoites of the malarial 
parasite in Anopheles barbirosiris. 

Schiiffner (1902) describes and illustrates as “Anopheles II” 
an anopheline which is apparently Anopheles barbirostris. He 
was unsuccessful in attempts to transmit malaria with this 
species. 

Stephens and Christophers (1906) do not include this species 
among those known to transmit malaria. However, Liihe (1906) 
states that Stephens and Christophers believe from the geograph- 
ical distribution of this species that it may be a carrier of malaria. 

Stanton (1912) was unable in eight trials to infect the Malayan 
strain of this species with the parasites of subtertian malaria. 

Anopheles sinensis is another oriental species closely related 
to Anopheles barbirostris. Its synonymy is as follows: 

Anopheles sinensis Wiedemann, 1828. 
Myzorhynchus sinensis Theobald, 1901. 


Anopheles jesoensis Tsuzuki, 1902. 
Myzorhynchus peditaeniatus Leicester, 1908. 


- 


390 The Philippine Journal of Science 1914 


This species is reported in the Philippine Islands by Banks 
(1906) and Ludlow (1908). In April of the present year we 
found it breeding in a rice paddy in Laguna Province. 

James (1902) states that he has not carried out any feeding 
experiments with this species. 

Tsuzuki (1902) infected Anopheles sinensis (jesoensis) with 
the parasites of tertian malaria, and transmitted the infection 
to a healthy person; he believes it to be intimately concerned in 
the transmission of malaria in Japan. 

Stephens and Christophers (1906) believe from its geograph- 
ical distribution that Anopheles sinensis is of little importance 
in the transmission of malaria. 

Stanton (1912) observed malaria zygotes in this species on two 
occasions in the Federated Malay States, but was unable to infect 
it under experimental conditions with the parasites of subtertian 
malaria. 

Anopheles maculatus is a species whose identity has been very 
uncertain. The synonymy of this species is given by Stanton 
(1912) as follows: 

Anopheles maculatus Theobald, 1901. 

Anopheles maculata Theobald, 1901. 
Nyssorhynchus maculatus Theobald, 1903. 
Nyssorhynchus willmori Leicester nec James, 1908. 
Nyssorhynchus pseudowillmori Theobald, 1910. 

According to Stanton (1912), who has examined the type 
specimens of Anopheles maculatus Theobald, this confusion has 
arisen from the fact that the types are not male and female of 
the same species, but represent distinct species, the male being of 
the species known to oriental investigators as maculatus and the 
female of the species known to them as karwari. Stanton pro- 
poses to obviate this confusion by retaining the name maculatus 
for the species which is now well known under that name to the 


students in the Orient and to rename the species which had hither- 


to been known as karwari. 
Alcock (19138) lists Anopheles willmori as a variety of Ano- 
pheles maculatus. 


This species has not hitherto been credited to the Philippine | 


Islands. Mr. Banks, entomologist of the Bureau of Science, 
states that he recently collected several specimens at Baguio, 
Mountain Province, at an altitude of about 1,500 meters, but 
the report of these has never been published. Anopheles macu- 
latus is reputed to be essentially a mountain or highland species. 
However, we have found it breeding to a limited extent in Laguna 
Province, at an elevation of only about 100 meters above sea level. 


= so 


ee ee ee 


a ae 


Wi 


1x,B,5 Walker and Barber: Malaria in the Philippines 391 


The larve were found always along the banks of densely shaded 
brooks, limited in distribution, and more plentiful during the 
cool season. 

Schiiffner (1902) attempted experimental infections at Deli, 
Sumatra, with an anopheline described by him as “Anopheles Ia,”’ 
which he considered a variety of his “Anopheles I” (Anopheles 
rossi). From his description and figure this species appears 
to be Anopheles maculatus. He was unable to get this species 
to bite or to suck human blood. 

Stephens and Christophers (1906) list Anopheles maculatus 
among the species that have been proved to be capable of trans- 
mitting malaria, but in view of the confusion existing at the time 
they write as to the identity of this species their statement is not 
conclusive. In their diagnosis of this species they state that 
the female of Anopheles maculatus has four white bands on the 
palpi, which does not correspond to the revised species of Stanton, 
but rather to Anopheles karwart. 

Stanton (1912) states that both he and Doctor Watson have 
found this species infected in nature, in the Federated Malay 
States, and that he has been able to infect it under experimental 
conditions with the parasites of subtertian malaria. 

One other species, Anopheles (Nyssorhynchus) fuliginosus 
Giles, has been bred from our collection of larvze, but not in suf- 
ficient numbers to determine its ability to transmit malaria. 
This species, however, is included by Stephens and Christophers 
(1906) among those capable of transmitting malaria; and Stan- 
ton (1912) states that he has infected Anopheles fuliginosus 
experimentally. 

II. METHODS 


This investigation was conducted at the Calamba Sugar Estate, 
Canlubang, Laguna Province, Luzon, about 60 kilometers from 
Manila. A moderate amount of malaria existed there among the 
Japanese and Filipino laborers. As malaria was not prevalent 
in Manila, Canlubang offered a satisfactory place for carrying 
on our experiments within a reasonable distance from the Bureau 
of Science. The officials of the Calamba Sugar Estate codperated 
freely in the investigation. We are especially indebted to the 
resident physician of the company, Dr. Isaac S. Diller, who not 
only made available to us the hospital and patients for infection 
experiments, but took us into his home and permitted us to use 
one of his rooms in which to set up our temporary laboratory. 

The Calamba Sugar Estate is situated in Laguna Province, 
Luzon, near the borders of a large lake, Laguna de Bay. Along 


392 The Philippine Journal of Science 1914 


the borders of the lake the land is low and is for the most part 
devoted to the cultivation of rice with abundant irrigation from 
the numerous streams flowing through it. At a distance of 
several kilometers from the lake, the land rises to an extensive 
undulating plain elevated from one hundred to several hundred 
meters. This region is a part of the old Friar lands, devoted to 
the cultivation of sugar for many years during the Spanish rule. 
During the insurrection, in the latter years of the Spanish domi- 
nation, much of it was laid waste and grew up to cogon grass or 
jungle. Since the American occupation it is rapidly being re- 
claimed and devoted to sugar cultivation again, both by the Fili- 
pinos in a small way and by several American and European 
companies on an extensive scale. The soil is fertile and un- 
usually well watered. - The region is crossed by numerous small 
rivers and brooks, many of which are fed by springs and flow 
throughout the dry season. These streams are peculiar in that 
they have in most cases cut deeply into the soil, forming deep 
ravines, and often into the underlying soft volcanic rock, form- 
ing canons. The banks of the streams are for the most part 
densely covered with jungle vegetation. The lake, the extensive 
irrigated rice fields, and the innumerable rivers and brooks in 
this country afford abundant and varied breeding places for 
mosquitoes and furnish unlimited collecting grounds for the 
species of anophelines that occur in this region. 

All of the mosquitoes used in our experiments have been col- 
lected and bred from larve. The species considered in this 
paper were all collected in Laguna Province on the Calamba 
Sugar Estate and adjacent country, with the exception of the 
strain of Anopheles rossii that breeds in brackish or salt water, 
the larvee of which were collected in esteros on the borders 
of Manila Bay, about 50 kilometers distant, and brought to 
Canlubang. 

COLLECTING LARV45 OF MOSQUITOES 


In collecting larve in the brooks and rivers, it has been found 
advantageous to wade the stream, preferably against the current. 
In this way the breeding places are more accessible, and it is 
often the only way to penetrate the jungle or to gain an entrance 
into the cafions peculiar to the region. 

A variety of methods were tried in collecting larve. At first, 
white porcelain evaporating dishes, about 14 centimeters in 
diameter, were used. By dipping these dishes into every sus- 
picious-looking pool, the larve, if present, were readily seen 
against the white background. When larve were found, the 


x,B,5 Walker and Barber: Malaria in the Philippines 393 


excess of water was first poured off. This was readily accom- 
plished, as the larve tend to swim against the current and collect 
at the opposite side of the dish. The remainder of the water 
with the larve was then poured into the collecting jar. How- 
ever, the method finally adopted as the most efficient was the 
use of white granite-ware pans, about 20 centimeters long, 13 
broad, and 5 centimeters deep, for dipping up the larve. The 
larvee were then removed from the surface of the water in the pan 
with a large spoon, which for convenience was carried attached 
to the finger or belt of the collector by a string, and were placed 
in the collecting jar. This pan provides a larger surface for 
skimming the surface of the water, while the corners are readily 
inserted into small nooks. The larve can be removed rapidly 
with the spoon from the surface of the water in the pan with 
a minimum excess of water. 

The collecting jar has usually consisted of a large wide-mouthed 
bottle of heavy glass, holding about one-half to three-fourths of 
a liter. One of these is attached to the belt of the collector by 
a cord about the neck, leaving both hands free to manipulate 
the dipping pan and spoon. On extended collecting trips it was 
necessary to take extra jars, which were carried by a boy ina 
hand bag. Occasionally these extra jars were replaced by a 
large 5-gallon can with a handle, which served as a storage 
receptacle for the larve. 

Various methods were made use of to get rid of the excess 
of water which accumulated in the collecting jars. One of these 
consisted of a collecting can with an inner cylinder of fine wire 
gauze provided with a bottom and extending above the top of 
the can. This permitted the escape of the water without loss 
of the larve. Another method was to siphon off the water 
from the bottom of the jar. The latter method accomplishes 
the end without loss of the larve if it is done from an undis- 
turbed jar in which the larve have collected at the surface. Two 
pieces of glass tubing connected by a short section of rubber 
tubing make a convenient siphon which can be carried in the 
pocket of the collector. 

From our experience in this investigation, we wish to em- 
phasize the advantages in experiments of this kind of collecting 
very large quantities of larve. Large numbers of the larve fail 
to develop and many of the adult mosquitoes die during the 
course of the experiments; therefore, time and labor will be 
saved ultimately if the collections of larve be made on as ex- 
tensive a scale as possible. 


394 The Philippine Journal of Science 1914 


BREEDING MOSQUITOES FROM THE LARV& 


The collected larvee were carried to the laboratory and placed 
with the water in the breeding jars. These consisted of battery 
jars, or large open-mouthed bottles, the tops of which were 
covered with netting secured by rubber bands. These jars were 
kept on the open veranda outside of the house. The sun-loving 
species were placed where they would get a certain amount of 
sunlight each day, and the shade-loving species were placed in 
a shaded position. Every morning the laboratory assistant 
aérated the breeding jars by passing a current of air to the 
bottom of the jar, as recommended by Darling (1910). A major- 
ity of the pupz and large larve always developed, but many 
of the small larvze failed to develop from lack of food or for 
other reasons. i 

When the pupz had hatched and the mosquitoes had collected 
in the upper part of the breeding jar, they were allowed to escape 
into the biting cages. These latter consisted of lantern chim- 
neys, aS recommended by Darling (1910). The upper end of 
the chimney was covered with mosquito netting fastened by 
a rubber band, and the lower end was set into the half of a 
Petri dish. The supply of a proper amount of moisture to sat- 
urate the atmosphere, but not drown or sprawl the mosquitoes, 
was a problem of considerable importance and difficulty. In open 
dishes of water placed in the cages, the mosquitoes were fre- 
quently drowned, even when a float of cork was placed on the 
surface. In our earlier experiments small stender dishes, filled 
with absorbent cotton over which were placed several layers 
of filter paper, were filled with as much water as the cotton 
would absorb. This served the purpose fairly well, but as the 
hot season advanced the heavy mortality among the mosquitoes 
led us to seek other methods that might prolong the life of 
the insects. In an attempt to imitate natural conditions, the 
Petri dish bottoms of the cages were filled with moist earth 
containing a growing plant. This proved to be little, if any, 
superior to the saturated cotton, and molds rapidly developed on 
the moist earth. Finally the bottom of the cage was filled with 
washed sand, which was kept saturated with water. This method 
proved to be the most satisfactory. Sufficient water was sup- 
plied, and the sand served as a resting place for the mosquitoes 
without danger of drowing; a large surface for evaporation was 
furnished which saturated the atmosphere without condensation 
on the surface of the glass, in which the mosquitoes become 
sprawled; and there was no difficulty from the growth of molds. 


1x,B,5 Walker and Barber: Malaria in the Philippines 395 


In these cages the mortality of the mosquitoes was low, even 
during the hot season. 

The cages were kept in a dark closet in the laboratory at 
room temperature. In order to keep out the ants, the legs of 
the closet were set in tins of petroleum. When not fed on blood, 
split raisins, placed on the netting covering the top of the cages, 
served as food for the mosquitoes. Every morning the cages 
were gone over and fresh water and food supplied. The cages 
were changed occasionally when they had become soiled. It 
was found that the mosquitoes bit better if no food was supplied 
before and between the feedings on blood. 


IDENTIFICATION OF THE SPECIES OF THE MOSQUITOES 


The species of each mosquito was determined when it was 
removed from the cage for dissection. All of the males and 
such females as died in every cage and the remains of every 
female dissected were preserved for confirmatory identification. 
These confirmatory identifications were made by Mr. Charles §S. 
Banks. 


THE SELECTION OF PATIENTS FOR INFECTING THE MOSQUITOES 


A routine blood examination was made of every patient en- 
tering the hospital at the estate. In addition, large numbers 
of blood slides were made of the laborers in the bunk houses 
on the plantation and of the inhabitants of the outlying barrios. 
Whenever a blood slide showed malarial parasites, it was 
examined especially for the presence of gametes, and if they 
were found in numbers that appeared to justify it a differential 
count of the gametes and leucocytes was made to determine if 
the gametes were sufficiently numerous for use in infecting the 
mosquitoes. When a patient was found to be suitable for our 
experiments, attempts were made at once to induce him to 
submit to the bites of our experimental mosquitoes. This was 
usually accomplished by a little persuasion and a small monetary 
compensation. The hospital patients were available at any 
hour for our experiments; gamete carriers from the plantation 
or outlying barrios reported to us outside of their hours of 
labor, either in the evening or early in the morning. Gamete 
carriers from outside barrios were several times taken wholly 
into our employ, and paid the same wages they would have 
received if employed at their regular work. Such carriers were 
not given treatment, unless they developed marked symptoms, 
until we were through with them. 


396 The Philippine Journal of Science 1914 


The number of gametes in the blood of patients was deter- 
mined approximately by making a differential count of gametes 
and leucocytes. In a few of the earlier experiments the gametes 
were not counted. Following these, differential counts were 
made on thin blood smears, from 200 to 500 leucocytes being 
counted in each case. In the latter experiments, which include 
all of the more exact quantitative experiments, the differential 
counts were made on thick blood smears, in which from 1,500 
to 3,000 leucocytes were counted. Darling (1910) has estimated 
that the limits of infectiousness of man to mosquitoes is about 
1 gamete to 500 leucocytes, or 12 gametes to a cubic millimeter 
of blood. No patient was intentionally used in which the gametes 
approached the limits of infectiousness as estimated by Darling, 
and an effort was made to obtain patients for our experiments 
with as high a gamete count as possible in order to increase 
the probabilities of infection of the mosquitoes. Every time 
the mosquitoes were fed on a patient, a blood smear was taken 
for a gamete count in order to have information of the number 
of gametes in the blood at the time of feeding. 

Some of the patients employed for the biting experiments 
were on quinine treatment. Darling (1910) claims, howeyer, 
that this does not affect the infectiousness of the blood for 
mosquitoes. This question will be considered at length in the 
discussion of our experiments. , 

Thomson (1912) found that in a case of subtertian malaria, 
treated with quinine until all of the schizonts were killed, the 
gametes persisted for about twenty-one days; that is, the ga- 
metes of Plasmodium precox (falciparum) are not affected by 
quinine and their life is about three weeks. This has not 
been wholly our experience. In tertian and quartan infections 
the gametes quickly disappear from the blood under quinine 
treatment, and frequently also without any treatment. On the 
other hand, the gametes of subtertian malaria are much more 
persistent. They have frequently persisted for weeks, whether 
or not the patient was under quinine treatment. Usually, 
however, the gametes after reaching the maximum gradually 
diminish in number whether or not the patient is being treated. 
Many of the gamete carriers showed few or no vegetative forms 
of the parasites in their blood, and frequently presented no 
symptoms over considerable periods of time. These were kept 
off quinine and used repeatedly for biting experiments. 


ae 


x, B,5 Walker and Barber: Malaria in the Philippines 397 


INFECTING THE MOSQUITOES 


The mosquitoes have, in most cases, been kept from twenty- 
four to forty-eight hours without food after emerging before 
use in our experiments. While at times they would bite well 
on the same day that they emerged, on the whole they bit 
better if kept without food until the second day. The time 
of feeding the mosquitoes on the malarial patients was for the 
most part in the early morning. The advantage of feeding the 
mosquitoes in the early morning, rather than in the evening, 
is that good light, necessary for separating the females that 
sucked blood from the empty females and the males, is available. 
Some attempts were made to induce them, by darkening the 
cage, to bite during the daytime, but with little success. 

In the biting experiments, the lantern-chimney cage was 
removed from the Petri plate and netting placed over the lower 
end. The sides and bottom of the cage were wrapped with 
black cloth, and the top, covered with netting, was applied to 
the moistened skin of the patient. By holding the gauze-covered 
end toward the light before applying it to the patient, most of 
the mosquitoes could be induced to collect on the netting, which 
was then applied to the skin of the patient. In most cases the 
arm, but in some cases the body, of the patient was used for 
biting. It was found convenient to place several cages in a 
row in a valise or box not deeper than the height of the cages 
to hold them in position, and then have the patient rest his 
arm on the tops of the cages. Three or four cages and some- 
times more were fed at one time on the patient. The cages 
were left in position until the patient felt no more bites or until 
it was considered that no more of the mosquitoes would bite. 
This was usually from fifteen to thirty minutes. 

After the feeding, the gauze on the lower end of the cage was 
- removed and the cage replaced over the Petri dish. Our lots 
of mosquitoes were allowed to bite the malarial patient on one, 
two, or three, or even more successive nights or mornings as 
circumstances or the purpose of our experiments demanded. 
During this period they were given no other food. After the 
feedings on infected blood were completed, the mosquitoes were 
supplied with raisins and sufficient water and were kept in the 
dark closet until ready for dissection. 

In the preliminary experiments to determine the capabilities 
of the different species of mosquitoes to become infected with 


the malarial parasites, no attempt was made to separate the 
129558——2 


398 The Philippine Journal of Science 1914 


\ 

mosquitoes that bit from those that did not bite the infected 
patient nor the males from the females. The mosquitoes were, 
when possible, given several opportunities to feed on blood on 
successive evenings, and it was presumed that the majority 
availed themselves of the opportunity. However, it is probable 
that a greater or less number of them failed on every occasion 
to suck blood. In the later experiments, in order to obtain an 
accurate comparison of the susceptibility of the different species 
proved to be capable of carrying malaria, the different species 
were fed simultaneously on the same gamete carrier and then 
the females full of blood were separated from the empty or 
doubtful females and the males. This was accomplished by 
feeding the mosquitoes in large numbers in the cages in the 
ordinary manner and then removing them individually from the 
cages in test tubes and examining them with a hand lens to 
determine whether or not they were full of blood. This was 
determined by the distinct red color of the abdomen of those 
which are gorged with fresh blood. The examination must be 
made shortly after feeding on the patient, as the blood becomes 
dark in color a few hours after it is ingested. The swollen 
condition of the abdomen is not a safe criterion, especially in 
the dark-bodied species like Anopheles barbirostris, because this 
condition may be due to engorgement with water or raisins 
used as food. Those full of blood were then liberated together 
in a fresh cage. The mosquitoes in the comparative cages con- 
taining the different species fed simultaneously on the same 
patient were then dissected on the same date to determine the 
proportion of infections and the relative number of odcysts in 
the infected individuals. 


DISSECTION OF THE MOSQUITOES 


In the experiments to determine the capability of the different 
species to become infected, the mosquitoes were kept alive from 
five to ten days after the last feeding on infected blood before 
dissection ; that is, long enough to obtain well-developed odcysts, 
but not until the sporozoites had escaped from the odcysts. The 
advantages of such early dissections for the purpose intended 
are that there is less loss of the mosquitoes by death and a 
considerable saving of time in the dissections, as it is only 
necessary to examine the mid-gut. The dissection of the salivary 
glands takes much more time and careful work. 

The mosquitoes to be dissected were killed in the cage by a 
few drops of chloroform. The males when present were re- 


ied 


Sil 


1x,B,5 Walker and Barber: Malaria in the Philippines 399 


moved, and each female, as she was removed from the cage for 
dissection, was carefully examined for the identification of the 
species, as it is not uncommon for two or more species to be 
represented in the same cage. When only the mid-gut was dis- 
sected, the thorax and head were preserved for later confirma- 
tory identification. 

The dissection of the mid-gut was performed according to the 
method described by Stephens and Christophers (1906). The 
mosquito is laid on a glass microscope slide over a white card 
on which is a blackened area. Over the white background, 
the abdomen of the mosquito is cut off at its junction with the 
thorax with a sharp needle. The abdomen is then moved over 
the black background, and a drop of physiological salt solution 
is placed upon it. The abdomen is held at the proximal end 
by one dissecting needle, while a second needle is pressed on the 
terminal segment and gentle traction exerted. With proper care 
and experience, the intestinal tract and the ovaries will be drawn 
out intact, attached to the terminal segment. If the hind-gut 
breaks off, leaving the mid-gut in the abdomen, it can be dis- 
sected out by slitting the wall of the abdomen with dissecting 
needles. The mid-gut is then cut off, and all of the Malpighian 
tubules are carefully removed with the needles, otherwise they 
tend to lie over the mid-gut and obstruct the microscopic view. 


_All of this dissection should be made under a simple lens sup- 


ported by a holder. A cover glass is then placed over the mid- 
gut, and it is examined first with the low power and then with 
the high-power dry lens of the compound microscope for oécysts. 
At the period of development at which our dissections were 
made, the odcysts could always be identified with a Zeiss DD 
objective, and usually with the AA objective. The immature 
odcysts of this age appear against the granular background of 
the gut wall, as round or slightly oval, transparent, structureless, 
feebly refractive bodies, 19 to 42 microns in diameter, and 
having a definite wall. Their identity is made certain by the 
presence of malarial pigment in the protoplasm. The older 
odcysts, in which the sporoblasts or sporozoites have developed, 
are larger, more granular, and darker in appearance, and the 
sporoblasts or sporozoites in them are readily recognized with 
the higher magnification. 

In order to determine whether the malarial parasites were 
capable of attaining complete development in the several species 
of the mosquitoes, it was considered necessary to give the para- 


400 The Philippine Journal of Science 1914 


sites an opportunity to develop sporozoites and infect the salivary 
glands. For this purpose, mosquitoes which had fed on infected 
blood were kept alive for from twelve to eighteen days before 
dissection. Both the mid-gut and the salivary glands were 
then dissected. In removing the salivary glands from the mos- 
quito, the methods described by Stephens and Christophers 
(1906) were employed. The legs and wings of the mosquito, 
the abdomen of which had already been removed for the dis- 
section of the mid-gut, are cut off with a sharp needle, the cuticle 
at the prothorax is torn with the needle, the slide is placed 
over a black background, the thorax is held by one dissecting 
needle, while gentle traction is exerted on the head with a second 
needle. This will, if properly done, draw out the salivary glands 
from the thorax attached to the head. They are then cut off 
from the head with a sharp needle, a cover glass is placed over 
them, and they are examined with the high dry and then with 
the oil-immersion objective of the compound microscope for the 
presence of sporozoites. In case of failure to remove the salivary 
glands by this method, they can still be found by carefully 
teasing apart the tissues of the anterior ventral part of the 
thorax. 

The mid-guts when found infected were preserved. In most 
cases the entire mid-gut was placed in 0.85 per cent sodium 
chloride solution containing 10 per cent formalin. In a few 
cases Schaudinn’s sublimate alcohol mixture was used as a 
fixative, after which the organs were preserved in 70 per cent 
alcohol. Smears were made of the infected salivary glands, 
which were stained by Giemsa’s stain, by sliding off the cover 
glass. Sections were not made for diagnosis, as the dissection 
method is much quicker and simpler; but in a few cases sections 
were cut for permanent preparations. 


Ill. EXPERIMENTS 


There is a notable lack of detail in the reports of most authors 
on their experiments in infecting mosquitoes with malarial par- 
asites. For this reason, and because our experiments have 
been more extensive and more quantitatively accurate that those 
hitherto performed, it has seemed desirable to give a detailed 
account of our investigation. In order first to present a com- 
prehensive view of our experiments in a form that will economize 
space and at the same time facilitate a comparison of the data 
and the results, their essential facts are tabulated in Table II. 


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Toten eens == MAMOMIQLDG * VY. 


The Philippine Journal of Science 


416 


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ix,B,5 Walker and Barber: Malaria in the Philippines 4?1 


In the text and tables that follow, the details of these experi- 
ments are discussed at length under appropriate headings. 


THE DEVELOPMENT OF ANOPHELINE LARV4 IN CAPTIVITY 


The rearing of adult anopheline mosquitoes from mature or 
nearly mature larve and from pupz in the laboratory presents 
no difficulties, as the majority of them live to emerge as imagoes 
within a few days; on the other hand, large numbers of the 
younger larvee fail to develop and die in the breeding jars. This 
is possibly due in part to the stagnant condition of the water in 
the breeding jars and in part to the lack of proper food. The 
water in our breeding jars was aérated daily, as recommended by 
Darling (1910), and no difficulty was experienced with foulness 
of the water due to the development of anaérobic bacteria. In 
collecting larvee, more or less of their natural food was undoubt- 
edly included, and in many cases alge, pond weeds, or débris, 
in which the larve were found, were added to the breeding jars. 
Duck weed, which in some cases was found associated with the 
larve in nature, when added to the breeding jars quickly over- 
grows the surface of the water and prevents the larve from 
coming to the surface for air. It is probable that the develop- 
ment of a larger proportion of the younger larve would be 
secured if they could be bred in the tanks furnished with run- 
ning water and supplied with an abundance of natural food. In 
practice it was found best to collect, so far as possible, the larger 
larve and pupe. 


PROPORTION OF FEMALES TO MALES IN ANOPHELINES BRED FROM LARV4 IN 
CAPTIVITY 


This is best shown in a table constructed from the data of those 
experiments in which the males and females were separated and 
counted. The counts were made of the mosquitoes after feeding 
on the malaria patient in the cages in which they were collected 
from the breeding jars, and include the dead as well as living 
mosquitoes. They are, therefore, approximately accurate. 


TABLE III.—Proportion of females to males in anophelines bred from larve. 


Lots of 
Species. mosqui-| Males. |Females.}Females. 
toes. 
ee ary 

Per cent. 
Anopheles febrifer -.....--.---------------- porte 8 Oe 35 | 697 812 53.8 
AR ODRELES TOBE ea en en te ie Se eS la a 37 787 1, 073 57.6 
Anopheles barbirostris__.__-__-_---- Sst 4a ae ee ee 23 206 289 58.3 
AIO DELES SULETISTS Se ete eee ee eee ee o> 4 96 143 59.8 
Anophelesimaciulatiss.. 5925. saae oe os eee oe = Bee 1 3 2 40.0 
NUD nt Ek red se RR AR el 100| 1,789 | 2, 319 | 56.4 


422 The Philippine Journal of Science 1914 


From this table it is apparent that the females outnumbered 
the males in every species but Anopheles maculatus. However, 
in this species the number of mosquitoes is too few to draw any 
conclusions. Authors have sometimes complained of a prepon- 
derance of males among mosquitoes bred from larve in the 
laboratory, and it has been supposed that this preponderance was 
due to lack of sufficient food in the breeding jars. As Table 
III shows, we have not experienced this difficulty; in our lots 
of laboratory-bred mosquitoes of 5 species, the males and fe- 
males have varied more or less in the different lots as is to be 
expected, but on the whole there has been a slight preponderance 
of females. 

THE LONGEVITY OF ANOPHELINES IN CAPTIVITY 


We have succeeded in keeping a fair proportion of our mos- 
quitoes alive from five to eighteen days, a time sufficient for the 
development of the malarial parasite to the odcyst or sporozoite 
stages. The different cages of mosquitoes showed considerable 
variation in this respect, even in the case of the same species, 
reared and kept under apparently the same conditions. At the 
beginning of our investigation, during the cool season, but little 
difficulty was experienced in maintaining a fair proportion of 
mosquitoes alive to the time of dissection. As the hot season 
approached, the mortality occurring among our experimental 
mosquitoes increased. The use of wet cotton and filter paper as 
a bottom for our cages was therefore discarded, and after ex- 
perimenting with moist earth and other materials moist sand 
was finally substituted, which proved successful in maintaining 
the life of the mosquitoes in captivity. Undoubtedly, if our 
mosquitoes had received unlimited personal attention, a larger 
proportion would have survived, but owing to the pressure of 
work the care of them had to be intrusted to the laboratory boy, 
who was also overworked and necessarily neglected them at 
times. Experiments 95 to 227a, in which both the number of 
mosquitoes that fed on blood and the number that lived to be 
dissected were determined, supply the following data bearing on 
this question. In all cases the mosquitoes had emerged from 
twenty-four to forty-eight hours before biting, and were con- 
sequently from 1 to 2 days old when the experiment began. 


x, B,5 Walker and Barber: Malaria in the Philippines 423 


TABLE 1V.—The longevity of anophelines in captivity. 


| 
| . 
| Mosquitoes Mosquitoes Mosquitoes 


* Paar that sur- 
. . - at begin- | surviving 
Interval in days between blood meal and dissection. minnie atidatelot vived and 


periment. | dissection. ee dis- 


Per cent 

Epa ae Se EN, A ee eee 11 | 9 81.8 
HOG Heme enemies sel Nees MT) Pt 158 121 76.5 
ee ee ee 139 69 49.6 
fo Ae Sak ous ee te ae SO a 188 80 42.5 
|G) ctl a ee I le a a Se eae A 298 151 50.6 
Open see teen. Suede ST) Se he td 100 47 47.0 
iif auiso wt Bud aS VER eet Re es ae Ce ces eee 112 32 28.5 
[NRE SAP ag eS PA AISNE) eee 120 64 53.3 
115 pee hart lle OS Ren re tse Ries ces eee tien ee Be Ae NE 32 19 59.3 
| Tisha oie SING EE 12 2) (OED REY Oe ee emeees BY TERI 8) 17 8 47.0 
0 TG ee Ae cea ee Bes eae ee eee eae aa 54 17 81.4 
| 1G Gest 2h eS ey Sven ais h 8 Weer bien ts eee oe 24 14 58.3 
ata atk Eola Rs ae alee parts nally oem =| 8 21.0 
RAR ReCORie ern ss - tka eS FREES _ See ee Oe ee 11 0 0 
Spoperee hiss “ated it panes CED alee ay Oe 1,302 | 639 49.0 


Therefore, on the average, about one-half of our experimental 
mosquitoes lived to be dissected. The proportion that lived to 
the date of dissection, so far as the differential counts of both 
the females that sucked blood and the females that lived to be 
dissected were made, show the following distribution among the 
several species: 


TABLE V.—Longevity of the several species in captivity. 


aces j aE 2 
Females | Females | Females 
Species. that sucked; aliveat living to be’ 
blood. dissection. | dissected. 
| Per cent. 
PARI ODER SIT EUT TER Ce tone oe na nn a ee 9 BA al 398 219 55.0 
PAWL OTINGLRMIT ORE hee we gee ee ee eR! ee 523 221 42.2 
PAMOMRELOSI ORY USTOSET ES) =. eee j 221 | 137 61.9 
PAR OTIEIDS SEIATEGtH Lees = eee Soets Felt Oke ee oS 23 18 18.2 
WeAnophetsaannculatud. 6 atone oe Mi ok Se eo | 2 2 100.0 | 


In this table are included all of the mosquitoes that lived to be 
dissected without reference to the length of time that they 
survived. The time intervening between the date that the mos- 
quitoes fed on infected blood and the date they were dissected 
probably averages nearly equal in the several species. 


AYA The Philippine Journal of Science 1914 


The data are probably approximately accurate for the first 
3 species, but in the last 2 the numbers are too small for the 
percentages to be reliable. 


THE AVIDITY OF THE SEVERAL SPECIES OF ANOPHELINES FOR HUMAN BLOOD 


In experiments 1 to 94, inclusive, the females which contained 
blood were not separated from the empty females and males 
after feeding on infected blood; the mosquitoes in this series 
were usually given two or more opportunities to feed on the 
malarial patient on successive days. It is, therefore, impossible 
to say what proportion of the mosquitoes in this series fed on 
the infected blood at any one time. In experiments 95 to 227a, 
inclusive, the mosquitoes, which were given an opportunity to 
bite the infected patient only once, were immediately removed 
individually from the biting cages in test tubes and examined 
with a hand lens. The females containing blood were separated 
from the empty females and the males, and were placed in a 
separate cage which was given the experiment number. The 
number of females containing blood, empty females, and males 
were, in most cases, recorded in this series of experiments. In 
Table VI are given the results of these differential counts, clas- 
sified according to the species of Anophelinze. Some of these 
mosquitoes were fed on the malarial patient within twenty-four 
hours, others not until the second day after emergence. In 
the latter case, all food was usually withheld from the mosquitoes 
until they were fed on the malarial patient. 


TABLE VI.—The proportion of the different species of anophelines taking 
a meal of blood when given one opportunity. 


Females Taking a 
“1 Total Eas Empty 5 
Species. containing meal of 
females. Blood: females. Rica 
| 5) eee ee y: Neel beet eT) 
Per cent. | 
Anopheles febrifer--------- ih hie Rue cee 696 382 314 54.8 
BMOTRELER IT OSE. 06 oo oe re ae coe ee on 936 596 340 63.6 
|| CAMO DAles DAT DITORUL IS eens eee en ee 273 135 138 49.4 
LAL OMIVALOB RUILOIIS ER a ot ee ee a eee 59 40 19 67.8 
Anopheles maculatus.__.-.2-.--------~-----.----- 2 nf 1 50.0 
[ pes pa ate ae oe Oe Ae 1, 966 | 1, 154 812 | 58. 69 


From these data it appears that, as a whole, slightly over half 
of our mosquitoes took a meal of human blood when given one 
opportunity from twelve to forty-eight hours after emerging 
from the pupz. In some instances the mosquitoes bit well on 


rae ne ee ye ee 


1x,B,5 Walker and Barber: Malaria in the Philippines 425 


the same day that they emerged, but in general they bit better 
on the second day after emergence. With reference to the sev- 
eral species, while the differences are not pronounced, Anopheles 
sinensis shows the largest percentage of individuals taking a 
meal of blood, followed in order by Anopheles rossii, Anopheles 
febrifer, Anopheles maculatus, and Anopheles barbirostris. 


MALARIA PATIENTS USED TO INFECT THE MOSQUITOES 


Seventeen different malaria patients whose blood contained 
gametes were used for infecting the mosquitoes in our experi- 
ments. Of these, 8 were suffering with subtertian, 7 with ter- 
tian, and 2 with quartan malaria. More experiments were made 
with subtertian malaria, because this type was most frequently 
met with and because the gametes persist in subtertian malaria, 
while they quickly disappear from the blood in tertian mala- 
ria, especially if the patient is on quinine treatment. Quartan 
malaria was found only rarely, and consequently only a few in- 
fection experiments were performed with this type of malaria. 
Thirteen of the patients were Filipinos, of whom 11 were males 
and 2 females; and 4 were Japanese, all males. The number 
of cages of mosquitoes fed on one patient varied from 1 to 32, 
and depended upon the persistence of the gametes in the blood 
and the ability to keep the patient in our service. 


NUMBER OF FEEDINGS ON INFECTED BLOOD 


The mosquitoes were allowed to feed from 1 to 6 times on 
the infected patient in the different experiments. In the earlier 
experiments the number of feedings depended upon the supply 
of mosquitoes and the availability of a gamete carrier; in experi- 
ments 95 to 227a, the mosquitoes were fed only once, when 
those containing blood were separated from the empty females 
and males. It is probable that a greater number of feedings 
on infected blood would increase the proportion of infections; 
but in these later experiments it was our purpose to obtain 
comparable results, and this was possible only under uniform 
conditions of experimentation. 


THE GAMETE COUNT IN MALARIA PATIENTS USED IN THE EXPERIMENTS 


In experiments 1 to 10, 17 to 19, and 29 to 33, the gametes 
were not counted but were estimated as +, ++, +++; in 
experiments 11 to 16, 20 to 28, and 34 to 55, inclusive, a differ- 
ential count of gametes and leucocytes was made in thin blood 
smears, from 200 to 600 leucocytes being counted; in experi- 
ments 56 to 227a, the differential counts of gametes and leuco- 


426 The Philippine Journal of Science 1914 


cytes were made in thick blood smears, from 1,500 to 3,000 
leucocytes being counted. In every case the gamete count is 
expressed in percentage of the leucocytes in the blood, and the 
counts were made from blood preparations made at the time 
the mosquitoes were fed on the patient. 

The counts range from 0.6 to 87.0 per hundred leucocytes. 
This would equal approximately from 42 to 6,090 gametes to 
1 cubic millimeter of blood, basing the estimate on an average 
of 7,000 leucocytes per cubic millimeter of blood. In all cases 
this would be well within the limits of infectiousness for the 
mosquitoes as estimated by Darling (1910). In three experi- 
ments (50, 51, and 53), owing to the fact that the mosquitoes 
were fed on the patient at the same time that the blood speci- 
mens were taken and before the gamete counts were made, it 
was found that the gametes had disappeared from the blood. 
However, in two of these experiments the mosquitoes had pre- 
viously been fed on the same patient when his blood contained 
gametes. 

The gametes being the only stage of the malarial parasite 
capable of infecting the mosquitoes, it would be supposed that the 
percentage of infections, or at least the intensity of the infec- 
tions, would vary with the number of gametes in the infected 
blood on which the mosquitoes fed. In Table VII are given 
the percentage of infections and the average number of odcysts 
to an infected mosquito, classified according to the gamete count 
of the patient on which the mosquitoes fed. In this table are 
included all of the mosquitoes dissected. 


TABLE VII.—Relation between the number of gametes in the blood and the 
percentage and intensity of infection of the mosquitoes. 


Total | 
FS , Average 
Mosqui- Mosqui- eee of Infected | odcysts to 
Gametes in the malarial blood. toes dis- toes in- inf jet mosqui- jan infected 
sected. fected. ait toes. mosqui- 
qui- an 
toes. a 
Per cent. 
O-diper'cent:.-=..:-=-22 5-0-5. 556 86 494 15.5 5.7 
B10 peritentiesso 5 ee cast see 364 45 329 12.3 7.8 
10-15 per cent\< 22. ose i -h25- Sees 105 12 39 11.4 3.2 
15-20 ‘per cent)o<.. 22.2 eae ee 5 5 185 100.0 37.0 
20-25 per cent 222.25 22<. fee sees sees 27 18 622 66.6 35.1 
25-80 per cent. Wso25 be ee 23 10 200 43.4 20.0 
80-35 per cent 5.02 5- oe eee anaes 32 6 153 18.7 25.5 | 


76-80 per'cent -=.- 25-2 osce ees sosoeaek 22 9 196 40.9 7 arg 
80-90 percent ef. se Eee eS 22 1 4 4.5 4.0 
| No record: <<< === -c2sSesseece acts 135 5 11 3.7 gu 


Jn the first and third numbers in this column, in 4 and 6 of the mosquitoes, respectively, 
the salivary glands only were infected at the time of dissection. Each of these was recorded 
as having 1 odcyst in the mid-gut, but it may have had more. 


ie a a 


1x,B,5 Walker and Barber: Malaria in the Philippines 427 


From this table it is apparent that given a malarial patient 
whose blood contains gametes above the limits of infectious- 
ness, the percentage of infections and the intensity of infections 
do not depend alone upon the number of gametes ingested, but 
some other factors must be involved in their determination. 

James (1902) calls attention to the question of the maturity 
of the gametes as bearing on the infectiveness of the blood for 
the mosquito. He believes that the maturity of the gametes 
should be determined at the time of every infection experiment, 
by drawing some of the patient’s blood on a microscopical slide 
under conditions that induce exflagellation of the microgameto- 
cytes in order to estimate the proportion of the mature gametes. 
We have not attempted this, believing that if the blood contained 
gametes in number well above the limits of infectiousness at 
least a part of them would be mature. Variations in the matur- 
ity of the gametes may be one of the factors that would account 
for the irregularities in Table VII. 


THE AMOUNT OF QUININE ADMINISTERED TO THE MALARIAL PATIENTS USED IN 
OUR EXPERIMENTS AND ITS EFFECT ON THE INFECTION OF THE MOSQUITOES 


Darling (1910) states that in his experiments the routine 
ward treatment with quinine, grains 10 ter in die, in solution 
apparently had no effect on the parasites and their develop- 
ment in the gut of the mosquito. However, he cites one experi- 
ment in which the patient had received no quinine for several 
days before the mosquitoes were fed and none during the experi- 
ment. In this experiment, one mosquito of the very suscep- 
tible species, Anopheles albimanus, showed a very large number 
(168) of zygotes in its mid-gut and 2 mosquitoes of the very 
unsusceptible species, Anopheles pseudopunctipennis, became 
infected. Therefore, he thinks that quinine may have a slight 
inhibitory effect on the parasite in the mid-gut of the mosquito. 
It would seem possible, although the gametes of the malarial 
parasite are resistant to quinine, that if the patient was saturated 
with the drug sufficient free quinine might be taken up with 
the blood into the stomach of the mosquito to injure or kill 
the delicate microgametes or the zygotes developed in the gut 
of the mosquito. 

In Table VIII are given the total amounts of quinine received 
by each of the patients during the period they were used in 
our experiments, the average amount per day, the percentage 
of infected mosquitoes obtained, and the average number of 
oécysts in the mid-guts of the infected mosquitoes in each case. 
The quinine was administered in the hospital, except in the 


428 The Philippine Journal of Science 1914 


cases mentioned in the footnote, and consequently the patients 
are known to have received the doses stated in the table. 


TABLE VIII.—Hffect of quinine treatment on infection of anophelines with 
malarial parasites. 


No, ot |Days useal ,Quining, | intraver | ‘Guinme |M@SMUtOe tomy | AUSERES, 
tie mt aaa during ex- = ples dunitioed- that fed infected. an infected 
periments. iments. | periments. on patient. | mosquito. 

Grams. Grams. Grams. Number.|Per cent. 

808 17 5. 83 9.0 0.85 60 0 0 0 
1251 6 0.64 7.2 1.3 hea 0 0 0 
1372 3 0 0.9 ET Hee ey ae 0 0 0 
1413 3 0.97 0.9 0. 62 22 1 4,54 1.0 
1261 13 5.18 9.0 1.09 82 5 6.09 37.0 
1486 3 0 0 0 12 0 0 0 
1506 2 0 0 0 12 1 8.3 3.0 
1845 2 0 0 0 21 2 9.5 3.0 

223 22 6. 80 10.8 0.8 271 21 Tem 11.5 
1240 3 2.59 0 0. 86 15 1 6.6 1.0 
1548 3 (3) 1.8 0.6? 19 2 10.5 2.5 
1612 16 0.97 0 0. 06 256 46 17.9 63.09 
1197 6 9.7 0 1.62 96 0 0 0 
1559 5 1.29 0.9 0, 44 57 0! 0 0 
1312 5 ©4, 86 0 0.97 252 74 29.3 bD5.7 
1786 2 0 0 0 44 10 22.7 20.0 
1806 7 9. 07 0 1,29 83 34. 40.9 28.8 


® Given quinine to take at home. 

> Several of the infected mosquitoes had only sporozoites in their salivary glands; therefore, 
they had at least 1 odcyst in the mid-gut and possibly more. 

© Given quinine to take at home. The patient said that he took the amount stated. 


From these data it does not appear that the quantity of quinine 
taken by the patient during the time he was used for infection 
experiments had any influence upon the percentage or intensity 
of infections of the mosquitoes. 


NUMBER OF EXPERIMENTS 


Anopheline mosquitoes, bred from larve, distributed among 
227 cages were fed on malaria patients whose blood contained 
gametes of the malarial parasite. Some of these cages were 
combined, after the mosquitoes had fed, to form the 184 experi- 
ments recorded in Table I. In our earlier experiments our 
technique was not fully developed, difficulties in keeping the 
mosquitoes alive were encountered, and the results were not 
quantitative; later our technique was improved, the difficulties 
were overcome, and the experiments were made to yield quanti- 
tative results. It has, however, been considered advisable to in- 
clude all of our experiments, the failures as well as the successes, 


IX B, 5 Walker and Barber: Malaria in the Philippines 429 


in our report. This will indicate better the reliability of our 
work, and will be helpful to others who may undertake similar 
investigations. 

Of these 184 experiments, 79 were with Anopheles febri- 
fer, 117 with Anopheles rossi, 59 with Anopheles barbirostris, 
5 with Anopheles sinensis, and 19 with Anopheles maculatus. 
Each cage contained a variable number of mosquitoes, and many 
of the cages contained two or more species, just as they had 
emerged from the larve collected in one locality. 


NUMBER OF MOSQUITOES DISSECTED 


In the 184 experiments, 1,287 lived to be dissected for the 
presence of odcysts in their mid-guts; of these, the salivary 
glands were also dissected in 316 for the presence of sporozoites 
although as shown in Table V about 50 per cent of the mos- 
quitoes died during the course of the experiments. These totals 
are distributed among the several species as follows: 


TABLE IX.—Distribution of the dissected mosquitoes among the several 


i species. 
c Mid-guts ENE 
Species. . lands 
| dissected. dissected.| 

PAT OMNeleRepTufer = ae ee ot ee ee Pee eee ge: Seed RS Se SR 373 93 
BAEEYT LEAST ORE ree ae Pe rey em en i et he a Sg 642 129 
ESIRIEITNE LE? OCS SE DSUERE 2a ne 5 ne ee ee. ee ee 205 10 
POTEET Oe eee re ee ee 2 ee oe ee ee ee 18 0 


DATEAREY ELAR NTL CULLIE IES © oe oa te ee ee tae On See a ORI S eo 49 24 
TREO i cee ae PLA IRA 9 Ai LE SS 7s Me a AN er a 1, 287 | 316 
— I _ 


DEVELOPMENT OF OVA IN THE EXPERIMENTAL MOSQUITOES 


Of the 1,287 female anopheline mosquitoes dissected, the ova 
were developed in 73, not developed in 1,139, and not recorded 
in 75. As in the case of the mosquitoes in cages 95 to 227a 
the females were separated from the males shortly after emerg- 
ing, it is fairer to exclude these from consideration, as the 
opportunity for copulation was greatly reduced. Of the 630 
females dissected from these cages only 26 showed development 
of the ova. In the remaining cages, 1 to 94, the males remained 
with the females up to the time of dissection. Six hundred 
fifty-seven females were dissected from these cages, of which 
the ova were developed in 47 and not developed in 492. No 
record was taken of the remainder. The distribution of the 
females having ova developed among the several species is shown 
in Table X. 


129558——4 


A430 The Philippine Journal of Science i914 


TABLE X.—Development of ova in the experimental mosquitoes. 


a Eercent- 
va not | age hav- 
Species. ove oS de- ing ova 
" ped. | veloped. de- 
veloped. 
AMODRELES FEOTISET, —— = xe Poa ee RE ee oe eee 9 381 2.3 
1 Anopheles rosavist Stoo ee 8 Se ER Se oe OL Bee See eS 17 541 3.0 
Anopheles barbirostyviss. --525 sos se ee ee ee ee 43 154 27.9 
Anopheles sinensis. o2 302) 0S eee ee ee eee 0 18 0 
Anopheles maculatwse a son nee ee ee ee 4 45 8.0 
| Total _...2.- 4/00 nite een pe Me ee anes "8 | 1188) Nene 


Annett, Dutton, and Elliott (1901) performed a number of 
experiments on the relation between a blood meal and the 
development of ova in anopheline mosquitoes; and from these 
experiments they came to the conclusion that a blood meal was 
necessary for the development of ova in the female. It would 
seem that the converse of this might be true; that every female 
which had a blood meal and copulated with a male would de- 
velop ova. It was hoped that the presence of developed ova 
might supply an index of whether or not the mosquito had bitten 
a patient and sucked blood, but this does not appear to be the 
case. 

PERCENTAGE OF INFECTED MOSQUITOES 


Of the total 1,287 female anopheline mosquitoes dissected 
after having had one or more opportunities of taking a meal 
of blood containing gametes, 205, or 15.92 per cent, contained 
odcysts in their mid-guts, or sporozoites in their salivary glands, 
or both odcysts and sporozoites, depending upon the time elaps- 
ing between the date of feeding and the date of dissection. Of 
the 205 infected mosquitoes, 189 contained odcysts in their mid- 
guts in various stages of development, and 28 showed sporo- 
zoites in their salivary glands. In experiments 1 to 94, the 
females containing blood were not separated from the empty 
females and it was impossible to make any differentiation at 
the time of dissection, but in experiments 95 to 227a only the 
females containing blood were dissected; in this series the pro- 
portion of infected females should be considered separately. 
Furthermore, a still more reliable comparison of the infections 
in the several species will be obtained by considering separately 
certain parallel experiments in which the different species of 
mosquitoes were fed at the same time on the same patient, in 
which the females containing blood were separated, and in 


1x,B,5 Walker and Barber: Malaria in the Philippines A831 


which at least one mosquito in the series became infected. The 
fact that one mosquito in such a series became infected proved 
that the gametes were capable of infecting. In Table XI are 
given the total infections and the infections grouped according 
to the above divisions, classified by species. 


TABLE XI.—Percentage of infected mosquitoes. 


be Af | aroma] Art| spre 4ypame| eas 
otal dispected Co - ee 3738 642 205 18 49 1, 287 
Total infected: 
TN) oz) cites Re SIE A ee I ie su ae 132 68 7 0 3 205 
IRemCen tes ee ere eee at ee 35.38 9.81 3.41 0 6. 12 15. 92 
Blooded females not separated from 
empty females: 
Phssected (2.5 fete 138 380 93 0 46 657 
Infected— 
Nim bercses anaes tas ee 24 28 1 0 1 54 
IRericent=2 26.22 ee 17.39 7.36 1.07 0 Peal; 8.21 
Blooded females only: 
IONICS os LE eee 0 Pe ee 235 262 112 18 3 630 
Infected— 
iN Der! seo oa Ue ie 2 108 85 6 0 2 151 
Réricent ste eee eee 45.95 13.35 5.85 0 66. 66 23.96 
Strictly comparative experiments: 
Phigsected == foes ere! 162 187 100 12 3 464 
Infected— 
Number seas! aM ds 108 36 | 6 C 2 151 
JEDIT) NS eee ee Cee ee 66. 66 18.71 6.0 0 66. 66 32.54 


In the last section of Table XI, the experiments with the 
several species are strictly comparable and the results give 
a more accurate measure of the susceptibility of the first three 
species. In the last two species the number of individuals 
included in the data is insufficient to draw reliable conclusions. 

In 1910 de Vogel was able to infect ‘““Myzomyia rossw’” breeding 
in brackish or salt water, but not those breeding in fresh water. 
Banks (1907) also obtained infections of “Myzomyia ludlowit” 
breeding in brackish and salt water, but he made no experi- 
ments with those breeding in fresh water. As our experiments 
were made with Anopheles rossii breeding in both fresh and 
salt water, it is interesting to compare the susceptibility of the 
mosquitoes from the two sources. Five hundred ninety-five 
specimens of Anopheles rossii breeding in fresh water were dis- 
sected, of which 58, or 9.74 per cent, were infected; 42 speci- 
mens of Anopheles rossii which were bred in salt water were 
dissected, of which 5, or 10.63 per cent, were infected. There- 


432 The Philippine Journal of Science 1914 


fore, while much fewer of the salt water forms were dissected, 
the percentage of infections in the mosquitoes from the two 
sources was practically the same. 


INFECTIONS WITH THE DIFFERENT SPECIES OF MALARIAL PARASITES 


The number and percentage of infections obtained with the 
parasites of subtertian, tertian, and quartan malaria in the 
different species of anophelines are given in Table XII. 


TABLE XII.—Infections with the different species of malarial parasites. 


= = 
Species. A. Se i- A. rossii. eae la. ae esas Total. 
Plasmodium prxcox (falciparum): 
Experiments 222. 2-0 66 91 48 5 18 228 
Mosquitoes dissected____-_------ 320 546 181 18 45 1.110 
Mosquitoes infected— 
Wamber= 3a cee rae 132 57 7 0 3 199 
Per .cent- ese ee 41.25 10. 43 3.86 0 6. 66 17. 92 
Plasmodium vivax: 
experiments S423. 525- ee ae 3 17 5 0 1 26 
Mosquitoes dissected __.--____-_- 13 75 il 0 4 103 
Mosquitoes infected— | 
INMimber So eee ee eee 0 6 0 0 0 6 
Percent 25. eee 0 8.00 0 0 0 5. 82 
Plasmodium malarizx: 
Experiments s5---.----ceee ee 9 8 7 0 0 24 
Mosquitoes dissected ____________ 40 21 13 0 0 74 
Mosquitoes infected— 
Number. so ceesc tt ee 0 0 0 0 0 0 
Per cent ¢ 62538 snes es 0 0 0 0 0 0 


In this table the total number of experiments with the sev- 
eral species of mosquitoes exceeds the number of experiments 
recorded in Table II because many of the cages contained more 
than one species of mosquito. While nearly as many patients 
with tertian (7) as with subtertian (8) malaria were used 
in our experiments, a much smaller number of experiments 
was made and mosquitoes dissected in the case of the tertian 
type of malaria. This is due chiefly to the fact that the gametes 
are much less persistent in tertian than in subtertian malaria. 
In the case of quartan malaria only two patient were available 
for experimentation. Moreover, both the tertian and the quar- 
tan patients happened to be available either early in our investi- 
gation before our technique was well developed or during the 
time when unusual mortality was occurring among our mos- 
quitoes. Therefore, our data with reference to infections with 
the tertian and quartan parasites are not so complete as desired. 


sare d<)! 


a 


1x,B,5 Walker and Barber: Malaria in the Philippines 433 
INFECTION OF THE SALIVARY GLANDS 


Of the 332 mosquitoes in which the salivary glands were dis- 
sected out for the determination of infection, 21, or 6.32 per 
cent, contained sporozoites. The total percentage of infected 
salivary glands is of no particular significance, as it depends 
upon the time elapsing between the meal of infective blood 
and the date of dissection and upon the species of anopheline. 
Therefore, the distribution of the mosquitoes dissected for 

_the salivary glands and the percentages infected are classi- 
fied in Table XIII according to the species and to the time of 


development. 
TABLE XIIJ.—Infected salivary glands. 


Days after infective feeding. 
Species. Fj 
10 il 12 13 14 15 16 18 | Total. 
Anopheles febrifer: 
Dissected: 22/2). ols aia 0 | 8 21 | 14 5 15 37 11! 111 
Infected— 
Numbers). es eee 0 0 3 3 0 2 6 6 20 
Pericent ute ee 0 0 | 14.28 | 21.42 0 | 13.33 | 16.21 | 54,54 (18.01 
Anopheles rossii: 
IDISSEC ECU een ee rey eae Hi 6 382 54 | 8 10 2 6| 125 
Infected— 
NumberBa.so22 = sesh) t 0 0 0 0 0 0 0 0 0 
Pemcerities 208 ae 3 0 0 0 0 0 0 0 0} 0 
Anopheles barbirostris: 
Dissectedeiat abet. yak 0 7 18 1 13 | 22 16 3 15 
Infected— | 
Nm bere = eee 0 0 0 | 0 0 0 | 0 | 0 0 | 
Rericent) 2222 - sik ee. NMG 0 0 0 0 0 Oe ot | 
Anopheles sinensis: 
Missected( esse lw me 2 oe 0 0 0 0 0 0 1) 0 0 
Infected— ! | | 
Numpbersi2as Sys. ee 0 0 0 0 | 0 0 0 0 
Pericent 22 en c ee 0 0 0 0 0 0 0 0 0 
Anopheles maculatus: | 
Dissected! <4. 2s2e se. o22 3. 4 4 1 6 2 4 0 0 21 
Infected— 
Number’ reuse weve 3c. 0 0 1 0 ae 0) 0 Oller 2 
Pericenteoe ea4 es 0 0 |100. 00 0 0 0 0 0 | 4.76 


The earliest day after the infective meal of blood that sporo- 
zoites were found in the salivary glands was the twelfth, in 
both Anopheles febrifer and in Anopheles maculatus infected 
with Plasmodium precox (falciparum), the parasite of sub- 
tertian malaria. These infections were found during the warm 
season; in the cool season the period of development of the 


434 The Philippine Journal of Science 1914 


subtertian parasite up to the infection of the salivary glands 
of the mosquito appeared to be slightly longer—from thirteen 
to fifteen days. No infected salivary glands were found among 
the mosquitoes dissected that had fed on the blood of patients 
infected with tertian or quartan malaria. For comparison with 
these results it is interesting to note that Darling (1910) found 
sporozoites of the tertian parasite after eleven and one-half 
days and of the subtertian parasite after eleven days in the 
salivary glands of Anopheles (Celia) albimanius in the Panama 
Canal Zone. 

It is noteworthy that, while 18.01 per cent of 111 Anopheles 
febrifer dissected between the tenth and eighteenth days after 
the infective meal showed sporozoites in the salivary glands, 
none of the 125 Anopheles rossiit dissected during the same pe- 
riods after the infective meal showed sporozoites in the salivary 
glands. However, odcysts containing sporozoites were found 
repeatedly in infected Anopheles rossi. 


IV. SUMMARY AND CONCLUSIONS 


Number of experiments, 184. Number of mosquitoes infected, 205. 
With subtertian malaria, 134. Mid-guts, 189. 
With tertian malaria, 26. Salivary glands, 28. 
With quartan malaria, 24. Anopheles febrifer, 132. 
With Anopheles febrifer, 79. Anopheles rossii, 68. 
With Anopheles rossii, 117. Anopheles barbirostris, 7. 
With Anopheles barbirostris, 59. Anopheles sinensis, 0. 
With Anopheles sinensis, 5. Anopheles maculatus, 3. 


With Anopheles maculatus, 19. Percentage of total mosquitoes in- 
Number of malaria patients used, 17. fected, 15.92. 


Subtertian, 8. Anopheles febrifer, 35.38. 
Tertian, 7. Anopheles vossti, 9.81. 
Quartan, 2. Anopheles barbirostris, 3.41. 
Gamete counts (percentage of leuco- Anopheles sinensis, 0. 
cytes), 0.6 to 87.0 Anopheles maculatus, 6.12. 
Number of mosquitoes dissected, Percentage of mosquitoes infected in 
1,287. the strictly comparative experi- 
Mid-guts, 1,287. ments, 32.54. 
Salivary glands, 316. Anopheles febrifer, 66.66. 
Anopheles febrifer, 373. Anopheles rossi, 18.71. 
Anopheles rossii, 642. Anopheles barbirostris, 6.00. 
Anopheles barbirostris, 205. Anopheles sinensis, 0. 
Anopheles sinensis, 18. Anopheles maculatus, 66.66. 


Anopheles maculatus, 49. 


The results of these experiments show that Anopheles febri- 
fer is probably, among the anopheline mosquito in the Philip- 
pine Islands, the most susceptible to infection with the parasites 
of subtertian malaria. While the number of experiments with 


1x,B,5 Walker and Barber: Malaria in the Philippines 435 


tertian and quartan malaria is insufficient to determine the 
fact, it is probable that this species is also an efficient carrier 
of these types of the disease. This species is from three to 
four times as susceptible as Anopheles rossii, which has hitherto 
been considered the malaria carrier of the Philippines, and 
eleven times as susceptible as Anopheles barbirostris. 

The number of mosquitoes of the species Anopheles sinensis 
and Anopheles maculatus dissected, especially in the compar- 
ative experiments, is too small to give reliable percentages. It 
is possible that a larger series of experiments with Anopheles 
sinensis would show that this species can be infected. Its 
susceptibility, however, is probably feeble. We believe it to be 
the least susceptible of the five species with which we experi- 
mented. The percentage, 66.66, of infections of Anopheles macu- 
latus in the comparative experiments was obtained with only 3 
specimens, and is probably too high; that of 6.12 in the total ex- 
periments is probably too low. We are of the opinion that the 
susceptibility of this species is at least as high as that of Ano- 
pheles rossii, and probably lies between that of the latter species 
and that of Anopheles febrifer. 

The réle played by a species of Anopheles in the transmission 
of malaria in any country depends chiefly upon (1) its suscep- 
tibility and (2) its geographical distribution and prevalence; 
also, to some extent, upon (3) its avidity for human blood and 
(4) its domesticity. 

Of these factors susceptibility is of fundamental importance. 
It is obvious that a mosquito which is immune or only slightly 
susceptible to infection with the malarial parasite will, no matter 
how prevalent or widely distributed, be of little or no importance 
in the transmission of malaria; on the other hand, a very sus- 
ceptible species may, although less prevalent, play a leading 
role in the spread of this disease. For example, Anopheles 
rossii was collected in native houses in certain regions in India 
by Stephens and Christophers and others in far larger numbers 
than was Anopheles culicifacies; but, while the latter species 
was found naturally infected with malarial parasites to the 
extent of from 4 to 16 per cent, not a single Anopheles rossti was 
found infected. These authors, therefore, concluded that Ano- 
pheles rossii, although more prevalent, played a subordinate, 
while the less plentiful Anopheles culicifacies played the chief, 
role in the transmission of malaria in these regions. 

A species that was rare or limited to certain regions or alti- 
tudes might, although very susceptible, be of little importance 
in the dissemination of malaria in a country as a whole. A 


436 The Philippine Journal of Science 1914 


species, like Anopheles ludlowii, which breeds for the most part 
near the sea coast, would be of little importance in an inland 
country; Anopheles maculatus, which is chiefly a mosquito of 
high altitudes, would be unimportant in a lowland country; and 
an anopheline that is not more or less adaptable to various con- 
ditions and altitudes would play a lesser rdéle in the transmission 
of malaria in a country of varied topography, vegetation, and alti- 
tudes. An anopheline must not only be susceptible, but it must 
be of wide distribution and prevalence to be of prime importance 
in the epidemiology of malaria in any country. 

Anophelines vary widely in their avidity for human blood. 
Schiiffner (1902) was unable to make any individuals of one | 
species with which he experimented bite and suck the blood of 
man; on the other hand, another species exhibited an intense 
voracity for human blood. It is obvious that an anopheline 
which has a strong inclination to seek and bite man will, other 
things being equal, be more apt to transmit malaria than a 
species which has less avidity for human blood. 

Certain species of Anopheline, like Anopheles rossi, may be 
called domestic species, although not to the extent of Stegomyia 
calopus or some species of the genus Culex, in that they breed 
in the open near human habitations; other species, like those of 
the Myzorhynchus group, may be termed wild mosquitoes, in the 
sense that they breed in and frequent the forest. Other things 
being equal, it is probable that the more domestic species would 
be the more important in the transmission of malaria, because 
of the greater opportunity offered to them to bite and suck the 
blood of malarial and healthy persons. However, in most trop- 
ical countries, especially outside of the cities, the importance 
of this factor would be more or less neutralized by the habits 
and customs of the natives. The natives of most tropical 
countries build their houses in or about the borders of the forest 
and near water, usually along the banks of the jungle streams, 
for the purpose of shade and other protection which the forest 
offers and in order to be near a supply of water. It is the 
custom, at least in the Philippines, for the natives to wash their 
clothes and to bathe in these jungle streams and also to carry 
water from them, often in the early morning or evening. There 
is thus every opportunity for these people to be bitten by the 
forest-loving anophelines that abound there. 

Much more work must be done, especially on the distribution 
and prevalence of the Philippine Anopheline, before the rdle 
of the different species in the epidemiology of malaria in the 


1x,B,5 Walker and Barber: Malaria in the Philippines 437 


Philippine Islands can be accurately determined. These aspects 
of the problem are now being investigated by one of us (Bar- 
ber), and already interesting and important results have been 
obtained, which will be published later. However, the impor- 
tance of the 5 species of Anophelinz, investigated in the trans- 
mission of malaria in the Philippines, can be roughly estimated 
as follows: 

Anopheles maculatus is probably a moderately susceptible, 
semiwild species, with a moderate avidity for human blood; but 
on account of its very local distribution it probably plays a very 
small part in the transmission of malaria in the Philippines, 
especially in the lowlands. It is said to be primarily a highland 
species, and if it should be found to be more prevalent in the 
mountain provinces it might prove to be of importance in the 
dissemination of malaria in those regions. 

Anopheles sinensis, has a low, if not negative, susceptibility. 
It appears from the literature, as well as from our own observa- 
tions, to be extremely localized in its geographical distribution 
in the Philippines; it is scare, and is a relatively “wild” species. 
Therefore, while the few experiments made with it showed 
this species to have a relatively high avidity for human blood, 
it is probable that its part in the transmission of malaria in the 
Philippines is negligible. 

Anopheles barbirostris stands the lowest in our experiments 
in its avidity for human blood, and it is a relatively “wild” 
species. It appears to have a wide but scattered distribution in 
the Philippines, and its susceptibility to infection with malarial 
parasites is rather feeble. On the whole, it is probable that this 
species plays a subordinate part in the spread of malaria in 
these Islands. : , 

Anopheles rossi is one of the most domestic of the anophe- 
lines, with a relatively high avidity for human blood. It is very 
widely distributed, especially along the coast and lowlands, and 
is relatively prevalent. Its susceptibility to infection with the 
malarial parasite is rather low. It is possible that this species 
may play a certain réle in the dissemination of malaria, espe- 
cially along the extensive coasts of this Archipelago. 

Anopheles febrifer is both a “wild” and also a domestic 
species in so far as shaded breeding places are afforded, with 
a relatively high avidity for human blood. It is by far the most 
susceptible among the 5 species investigated, and is probably 
the most susceptible species in the Philippines. If investiga- 
tions, which are now being carried on by one of us (Barber), 


438 The Philippine Journal of Science 1914 


prove it to be as widely distributed throughout the Archipelago 
as it is in Laguna Province, Anopheles febrifer is the most 
important mosquito concerned in the epidemiology of malaria 
in the Philippine Islands. 


LITERATURE CITED IN THE TEXT 


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™~ 


1x,B,56 Walker and Barber: Malaria in the Philippines 439 


STANTON, A. T. The anopheles mosquitoes of Malaya and their larve, with 
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Nore.—Specimens of Anopheles febrifer were sent to Dr. C. S. Ludlow 
at the Army Medical Museum, Washington, D. C. Doctor Ludlow kindly 
examined the specimens and reported by letter dated December 1, 1914, 
that the species is Myzomyia christophersi Theobold and is the same mos- 
quito as that which she has reported from the Philippines under the name 
of M. funesta Giles. (See Ludlow, Disease-bearing mosquitoes of North 
and Central America, the West Indies, and the Philippine Islands. Bull. 
No.4. War Department. Office of the Surgeon General. November, 1913. 
p. 36.) As Doctor Ludlow states in this footnote, “What the proper name 
for this species is, seems a little clouded.” Myzomyia christophersi (=M. 
listoni?) is a well known malaria carrier in the foothills of the Himalayas 
in India. 


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MALARIA IN THE PHILIPPINE GENERAL HOSPITAL, MANILA, 
P. I., DURING THE FISCAL YEAR 1913 


By Davin G. WILLETS 
(From the Biological Laboratory, Bureau of Science, Manila, P. I.) 


INTRODUCTION 


The object of this paper is to analyze the cases of malaria 
discharged from the Philippine General Hospital, during the 
fiscal year ending June 30, 1913. This analysis is based not on 
the results of a special study of malaria but on those of routine 
work. That this fact greatly affects the data submitted is 
apparent throughout the paper. I am responsible for the labo- 
ratory results adduced, while the credit and the responsibility 
for the clinical data rest with the clinicians. 


FREQUENCY 


A diagnosis of malaria was rendered in 348, or 5.17 per cent, 
of 6,732 cases discharged during the year; in 324, or 10.6 per 
cent, of 2,985 medical cases; in 15, or 2.09 per cent, of 717 obstet- 
rical cases; and in 9, or 0.3 per cent, of 3,030 surgical cases. 
It follows that 93.1 per cent, 4.3 per cent, and 2.6 per cent of 
the 348 infections occurred in medical, obstetrical, and surgical 
cases, respectively. 

According to the annual report of the Bureau of Health for 
the fiscal year 1913, a diagnosis of malaria was rendered in 6.1 
per cent of the admissions to Baguio Hospital, Baguio, Benguet; 
in 1.5 per cent to Bilibid Prison Hospital, Manila; in 10.2 per 
cent to Butuan Hospital, Butuan, Agusan (Mindanao) ; and in 
51.1 per cent to Iwahig Penal Colony Hospital, Palawan. 


ORGANISM PRESENT 


Of the 296 cases which were positive microscopically, 161, 
or 54.4 per cent, were infected with tertian; 150, or 50.6 per 
cent, with estivo-autumnal; 3, or 1.0 per cent, with quartan; and 
4, or 1.4 per cent, with unidentified parasites. Infection with 
both tertian and estivo-autumnal organisms occurred in 22, or 


7.4 per cent. 
441 


442 The Philippine Journal of Science 1914 


DISTRIBUTION OF CASES 


A. Geographic distribution—An analysis of the cases accord- 
ing to the apparent place of contraction of infection gave Manila 
35.6 per cent, Mindoro 17.5 per cent, Tarlac Province 9.8 per 
cent, Laguna Province 9.5 per cent, and miscellaneous provinces 
27.6 per cent. It is absolutely certain that the vast majority of 
the infections apparently contracted in Manila were in reality 
contracted elsewhere. As a matter of fact, while malaria is 
endemic in Manila it is not a common disease here. It is believed 
by some physicians that it is very rare indeed, because it is 
almost impossible to find an infection in an individual who has 
never been outside of the city limits. This same argument 
could, however, be applied to typhoid fever or any other disease 
which occurs here, as Manila occupies such a limited amount of 
territory that it would be rather difficult to find a child, to say 
nothing of an adult, who had not been recently beyond its 
borders into Rizal or Bulacan Province. 

The Bureau of Health’s report for 1913 for the entire Philip- 
pines, excepting Mindanao and Mindoro, gives a very different 
distribution of malaria in the Islands than is indicated in Table 
I. According to the clinical cause of death the distribution is, 
in part, as follows: 


TABLE I.—Distribution of malaria in the Philippine Islands. 


| 


| Percent- | Percent- | 


| age age 
Province. ea aa Bravince: ne aa 
malaria malaria | 
clinically. wera 
|] ec a 8 RO eS ee 
Ce Sh eee ee 27.6 


Ambos Camarines -__.--------_----.- 20.6 | 
18.6 | 
18. 4 
18.0 


Musgrave, Walker, and others! found malarial organisms in 
the blood of 34.06 per cent of 1,095 individuals in Mindoro, in 
February, 1912. 

B. Race, sex, and age distribution.—The race, sex, and age 
distribution of the cases is given in Table II. Fifty per cent 
of 66 Japanese, 4.8 per cent of 5,166 Filipinos, 4.8 of 352 
persons of miscellaneous race, and 4.2 per cent of 1,148 Amer- 
icans were cases of malaria. 


*This Journal, Sec. B (1914), 9, 187. 


IX, B, 5 Willets: Malaria in General Hospital, Manila 443 


TABLE I].—Race, sex, and age distribution. 


| Sex. Age. 


Num-; Per Male. Female. 4 Adults. Children. 
Race. ber. | cent. |_ cay 


Num-} Per |Num-|j Per |Num-| Per |Num-] Per 
ber. | cent. | ber. | cent.| ber. | cent. | ber. | cent. 


Bilipinowe. = cee 250} 71.9 188 | 75.2 62 | 24.8 218 | 87.2 32 | 12.8 


American/ 222.0522 242. =. 48 | 18.8 45) 93.8 3 6.2 45} 93.8 3] 6.2 

Japaneses... 202 25.25 33 9.5 BB LOONO) eee eee Bon OONO! | Sasa eee os 

Miscellaneous ___-.-_--_- au) 4.8 i BOON 0] ee cees eeeeeess 16] 94.1 Dag) 
Total ese base 348 | 100.0 283} 81.3 65 | 18.7 312 | 89.7 36 | 10.3 


a There are few foreizn women in Manila. 


LABORATORY EFFICIENCY 


IN FINDING MALARIAL ORGANISMS IN CASES CLINICALLY POSITIVE 


Seven of the 348 cases diagnosed were not examined in the 
laboratory. Of the remaining 341, 45 were negative and 296, 
or 85.1 per cent, were positive. In making the examinations, 
smears were made in the usual way and Wright’s stain was used 
exclusively. The 45 negative cases have been analyzed as 
follows: 

Two cases were positive in the laboratory when previously in 
the hospital. Each was given quinine after one negative exam- 
ination and later was discharged recovered. 

One was given quinine intravenously several hours before 
smears were taken. There was one laboratory examination; the 
patient was discharged recovered. 

Twenty-one were given no quinine in the hospital excepting 
iron, quinine, and strychnine to 2 of them. Six gave a definite 
history of having taken quinine prior to admission. Most of 
the 21 gave a history of more or less typical malarial paroxysms. 
Some were doubtless convalescent when admitted. Fifteen were 
discharged recovered and 4 improved. One died of acute bacil- 
lary dysentery, chronic malarial splenitis being found at autopsy. 
One was a surgical case, which had rises of temperature after 
operation, and which was discharged later, the febrile attacks 
having subsided without the administration of quinine. . 

Ten, in addition to the 6 already mentioned, had been taking 
quinine before admission. Hight of these were given quinine 
in the hospital after one negative; 1 of these was given quinine 
in the hospital after two negative examinations. One was 
transferred to another institution (San Lazaro) after two neg- 


444 The Philippine Journal of Science 1914 


ative examinations, no quinine having been given. Eight were 
discharged recovered, 1 improved. 

Eleven other cases were given quinine in the hospital. The 
records do not give the quinine history of these cases. Six 
were negative microscopically once; 3, twice, 2, four times. 
Seven were discharged recovered; 1, improved; and 1, unim- 
proved. Twodied. Autopsy in one case disclosed chronic 
splenic hyperplasia of. malarial origin, death being due to lobar 
pneumonia. Autopsy was not obtained in the other case. 

From a consideration of the foregoing facts it appears that 
failure to find malarial organisms in the peripheral blood of these 
cases may have been due to several factors, namely: 


1. Rendering a positive clinical diagnosis, upon scanty laboratory work. 
This is evidenced by the fact that 29 cases were examined once; 11, 
twice; 3, three times; and 2, four times. The practical question as to 
whether it is justifiable to delay treatment of suspected malarial 
cases until a positive laboratory diagnosis is made is here involved. 
In some cases it is justifiable and absolutely indicated, while it is quite 
contraindicated in others. The last word in an accurate diagnosis of 
malaria rests with the laboratory. 

2. Absence of malarial organisms from the peripheral circulation. This 
may have been a factor in the seventeen or more cases which had 
been taking quinine prior to admission to the hospital and, also, in 
those cases which were apparently convalescent from slight attacks 
of malaria. 

3. Errors in the laboratory. 

4. Errors in the clinical diagnosis. 


IN FINDING MALARIAL ORGANISMS IN CASES POSITIVE AT AUTOPSY 


Dr. B. C. Crowell, pathologist of the Bureau of Science, found 
evidences of malarial infection in 19, or 5.6 per cent, of 338 
autopsies performed upon individuals who died in the Philippine 
General Hospital during the year. Malaria was the immediate 
cause of death in 13, or 68.4 per cent, of the cases in which it 
was present and hence in 3.8 per cent of the total cases which 
came to autopsy. In 12 of the 13 cases the lesions were acute; 
in 1, chronic. The lesions were chronic in the 6 cases in which 
the immediate cause of death was not malaria, but bacillary 
dysentery in 1 case; nephritis in 1 case; bronchopneumonia 
in 1 case; lobar pneumonia in 2 cases; and peritonitis in 1 case. 

The Bureau of Health report for 1913 gives 26,138, or 14.2 
per cent, of 183,236 deaths in the Philippines, excepting Min- 
danao and Mindoro, due to malaria. 

Of the 19 cases, clinical diagnosis was made of 73.7 per cent. 
The 5 cases which were not diagnosed clinically were as follows: 
(1) The patient arrived at the hospital in an unconscious condi- 


. 


IX, B, 5 Willets: Malaria in General Hospital, Manila 445 


tion and lived one hour after admission, (2) the patient was in 
the hospital twenty hours, (3) the patient had severe jaundice 
and was in the hospital three days, (4) the patient died of peri- 
tonitis, (5) the patient died of bacillary dysentery. In the first 
3 cases acute, and in the last 2 cases chronic, lesions were found 
at autopsy. 

Of the 19 cases, 15 were examined microscopically ante mor- 
tem. The cases not examined were 1 to 4, inclusive, enumerated 
above. Of the 15 cases examined, 13, or 86.7 per cent, were 
positive. Each of the 2 negative cases was examined once, and 
chronic lesions were found in each at autopsy. Bacillary dys- 
entery was the cause of death in one, and lobar pneumonia in 
the other, case. Twelve, or 92.3 per cent, of the 13 positive 
cases were infected with zestivo-autumnal organisms and 1, or 7.7 
per cent, was infected with tertian parasites. 


MISCELLANEOUS LABORATORY FINDINGS 


Of the 296 cases which were positive microscopically for 
malaria, 275 were medical, 13 obstetrical, and 8 surgical; 243 
were admitted to the hospital primarily because of malaria, and 
32 for miscellaneous reasons, the malaria infection being detected 
in the routine examinations. Among the 243 admitted because 
of malaria, conditions were present in 58 which obviously ex- 
cluded them from consideration when endeavoring to establish 
the usual miscellaneous laboratory findings of malarial cases in 
the Philippines. The urinalysis, fecal findings, hemoglobin per- 
centages, and leucocyte counts of the remaining 185 cases are 
given in Tables III to VI, inclusive. 


TABLE III.—Urinalysis.* 


Examinations and findings. Number. | Per cent. 
Parsons notexamined co eee oe eee eet rf ae 
Prersone\examinedt 2a: 22 32-3 a a ee et ee eee Gy il hee a 
PemAON MINOR BLUE) saan oe te ee oe ee ee ee Fo oe avr rg eee al 
Persons with— 
Albuminie <2. sees et cece a eee tee eee ee ee ee 73 46.5 
A humin' and casts 2 coco soso oe ee et ee 38 24.2 
Albumin, casts, pus cells, and red cells ____________--___------------------ 13 8.3 
| AN umiinya nar once see ae es eee em ee yom SA Suen Ee 9 5.7 
Albuminiand)red) cells h2 2225s ae ee eae Beas ee a oe ed 8 5.1 
Albumin; castajand redi celle io) 222 a oe ea ee 8 5.1 
Albumin' caste; (andinus cellsis sees eese eee = se ew ane sae coe neue ce ans 3 L9 
| Albumin, pus cells, and red cells_________._.._________---_..--.-------__--- 2 1.3 
Otel ae eee a ae er a ee eee eee See a ae Lee 8 157 100.0 


Asa rule but one morning specimen of urine was examined. The heat and acetic acid test was 
used for albumin. 
129558——5 


AAG The Philippine Journal of Science 1914 


TABLE 1V.—Fecal findings. 


Examinations and infections. Number. | Per cent. 

Fe crasfeynls) 1G) Ae en GS eo ao Soo eer Se oe se oe ee PY | ee 

ersonsiexamined io. 6oi i. See ee eee ee ee as eee ABS) | sa 
Personsiinfected 2-35-55 022 Se ee aloe is al 0 otk ae 133 86.9 
IPersonsinerative sis. = ts 6a ee a EE oe oD 13.1 

Persons infected with— 

‘One apecies pete eee Se se ease aces 37 24.2 
‘Two species... othe SBR eR ee Se I ig Re eae 54 35.3 
Three species; 2325 <2 = =n 35 22.8 
Four BRGCIOD |. 5 = 22 sos oe ce Se eee see ee pe RE ape RG 7 4.6 
Total nooo oe ee eS Ce ee Oe REI oni 153 100.0 
Y (Pe 71 FN a Reh ate A as Mi a uate Sle A es i) ee 
Ascuris.f2 02.2 aks sss BoE Se Se oe eS ee eas 68 44.4 
Entlamoba' 22 Go 25502 eave Bos: Pee eS ee eee 49 32.0 
Hookworm 22.222-2-2-52 0255 2epc no oh ee ae ae oe eel 44 28.8 
Monad en <5. rio sa seeee ar ne ene oe eras oe ree eee aoe eee eee ee 14 9,2 
Balantidium . . 222s \so30 6 Hes eet ee ee Be ee ee ee 2 1.8 
Onpuris ocr a Base Se Se et ee oe eee 1 0.7 
Strongyloides Ao 15 ee Le ie, a ee = eee ee 1 0.7 
MP Gta ee Sek ke os ER Sh ee Se ee A ee 278 181.8 


The feces, as a rule, were examined but once, and the speci- 
mens were obtained after the administration of magnesium 
sulphate. Four cover slip preparations were examined of each 
case. It appears from Table III that 183, or 86.9 per cent, 
were infected with one or more species of intestinal parasites 
and that there were 278 infections found in the 153 patients, 
or 181.8 infections per 100 persons. 


TABLE V.—Hzmoglobin percentage." 


Examinations and percentages. Number. | Per aes 
(Persons not Gxamined 922. soncoc este once ces coen as eep tape canes Uae Eee Eanee 94) .coetore 3 
(Persons examined oi cic5. es see Soe wk aoe ceed cece cmee es eee 94, [ae wes 


Persons with percentage of— 


aEstimated by Tallqvist’s method. 


IX, B, 5 Willets: Malaria in General Hospital, Manila 447 


TABLE VI.—Leucocyte counts. 


| Examinations and counts. Number. | Per cent. 


Persons notlexairiinee ae = eer ne mete mi myeny PRT a ee ee ee EJ Ne teen ed 
Personaiexamimeds ts eset Lee eae ya Eee fee oe hae ee gh IN| ogee etre a 
Persons with counts, per c. mm., of— 
2600 tioie) SOG Berar, Sees es SO a se Ss ee Je ek 2 3 20 
BAKU SG Ss tel tI Re le a eae ate teh IO Sek ae ee 13 5.9 
SOOO) iar ne oe rece Re a Se Se ee es See 23 17.6 
BS CON artery ne ee er ele tN ee be 23 17.6 | 
COWS ET Dae oe aE See ee, eee ee ee ards Oe Cer Sener ee 26 19.8 
TAY EY th ee 8 oS SRS ee aren ae Se ret Parse Bee Dae ees ee 12 9.2 
OO E OLS GUN) earn eek ASS ad a Se ee ees Ce ee 12 9.2 
QTC 0 TOYO Se Sea Re Ree ae eee ee ey ee Gene peer as 6 4.5 
ROOOTOH10:900 so sess ob Red A sone seas Bee eee eee 5 3.8 
COUR Oni te GOO eee orcs cee Lie ge OL aay 2 se See AY Dee ei 5 3.8 
3 


12,000 TROL US 2 eae na os Re a eos a ee Senate omer eae ee 2.3 | 
| MUS UE ate ete sans a lias SMS BPs Aah de «UE TE 8 Coe UN I 131 00.0 

The counts were made as soon as possible after admission. 
As a rule one count was made on each case, a Turck’s counting 
chamber being used. 

If from 6,000 to 10,000 leucocytes per cubic millimeter of 
blood are accepted as normal, the approximate average count 
would be about 8,000. In this series of cases the average count 
was about 6,420 which, although within normal limits, is so 
near the lower limit that it represents a leucopenia. Sixty-two, 
or 47.3 per cent, had a definite leucopenia; 56, or 42.7 per cent, 
a normal count; and 13,’or 10.0 per cent, a leucocytosis. It will 
be recalled that cases with complications, which would obviously 
influence the leucocyte count, were excluded from this series 
and that the individuals in whom these infections occurred 
entered the hospital because of malaria. These results, there- 
fore, apparently represent the leucocyte counts in malaria for 
this locality. 

Thomson? found that the absolute count in malaria varies 
considerably. During sporulation, if it be slight (as occurs in 
latent or apparently cured cases), a leucocytosis is present, 
whereas, if it be marked (as exists during a definite paroxysm), 
there is almost invariably a leucopenia. Between paroxysms in 
cases of marked acute malaria the count increases up to or even 
beyond the normal count. These findings may explain some of 
the normal and abnormally high counts in the series of cases 
here reported. It would appear, however, that 52.7 is a rather 


*Ann. Trop. Med. & Parasit. (1911-1912), 5, 83; ibid. (1912), 6, 215. 


4A8 The Philippine Journal of Science 1914 


high percentage to be thus explained. Having noted the great 
frequency of intestinal parasitism and genito-urinary disorders 
in the Philippines, it was thought that perhaps a tabulation of 
the cases according to the fecal and urinary findings might re- 
veal the cause of many of the normal and the abnormally high 
counts. The results follow: 


TABLE VII.—F'zcal findings arranged according to the presence of a leuco- 
penia, a normal count, or a leucocytosis. 


Leucopenia. | Normal count. | Leucocytosis. 
Examinations and findings. Cases. 
Num-| Per | Num-| Per | Num-| Per 
} ber. | cent. ber. cent. | ber. cent. 
H ac ee a ee eee 
Rérdonsinecative:c-2-2 cena ft ot Ee 13 7 53.8 5 38.5 1 12% 
Persons infected with— 
| One speries shock ee a A eer ees 30 15 50.0 12 40.0 8} 10.0 
| Wwomkpeciess seo ae ee 41 16 89.0 22 53.6 3 1.3 
| Three !spectes —-4...0.= 52-9 s ee 29 14 48.3 11 87.9 4] 13.8 
| Pour bpeciedio. 222. ee ae 4 2 50.0 2 BOLO fo4 oy. Se 
i Total: 2 esse Se ee ae 117 54 46.1 52 44.5 11 9.4 
! ——— es] -——— ———| 
} A scar ie so Oe eee ae eco 56 27 48.2 24 42.9 5 8.9 
| Oyrohsires 5525 Sues ee ee eae 2th! 76 82 42.1 38 46.1 9} 119 
Hookworm <oe¢ 3.- Lena eae een eet: 35 15 42.9 16 45.7 4] 114 
YH a, Nae oes ee RN eS SEO oS 36 18 50.0 16 44.4 2 5.6 
Mond Sf) 2220) + ee eee ae 8 3 87.5 4 50.0 a Oo 
BOUlaAnhains = cep eee nesta l eee 2 Bly 1000 2k SS ee ee eee 
| Sirangulariees -A3e e e e Dl ee a ee AP TBONO Wot sf ae oe cee 
CERT AR oa ee ae eee 7A enh) She FSA al eS Pw Lai le se ere Lt Sees 
Air i ee aap Ney oe Magee od AOE 215 | 97 |, yi4.9 96| 44.4 22 | 10.2 
i ie 


TABLE VIII.—Urinalyses arranged according to the presence of a leuco- 
penia, a normal count, or a leucocytosis. 


Persons nerative._-. 2-08.22. 5822-4 Mitac. eletowetae i POO Olfece., £8 | Lic pied 


Persons with— 

Albumin. 2 2ss38 5 cece esosstaedaas 56 23 42.6 27 

| Albumin and red cells -___---------- 7 4 57.1 3 
Albumin and pus cells -__------_---- 4 Mt) WOOLOW E2652] 22.22 ee ee eee 
Albumin, pus cells, and red cells ___ 2 1 50.0 | it 
Albumin and casts-___.-_----------_- 29 14 48.3 12 
Albumin, casts, and red cells _______ 5 2 40.0 3 
Albumin, casts, and pus cells_______ 3 2 Ci Sy fe Wee Sees Kaya sae Oe 

| 


Albumin, casts, pus cells, and red 


Cells. ee eee eee ees ere 11 {{ 63.6 4 
Ig a A any Tie 118 57 48.3 | 51 | 43.2 


A study of these tables will convince one that neither the fecal 
nor the urinary findings alone nor the two combined are obviously 
responsible for causing the normal or the abnormally high 
counts. On the contrary, it would appear that each may have 


IX, B, 5 Willets: Malaria in General Hospital, Manila 449 


contributed to the leucopenia. The fecal findings among per- 
sons of foreign birth were excluded and those among Filipinos 
alone considered, but the general rules were not changed appre- 
ciably. It is not overlooked that the urinary findings were due 
doubtless to malaria itself in many of these cases. 

The cause of many of the normal and the abnormally high 
counts in this series of cases is, therefore, considered to be 
undiscovered. Despite the unconvincing evidence presented in 
the tables, I am inclined to hold intestinal parasitism and genito- 
urinary disorders responsible for some of them. 


SPLENIC INDEX 


To determine the splenic index of malaria, the records of the 
first 1,000 cases for the year in which a complete physical exam- 
ination had apparently been made were examined. In 799 of 
these cases the spleen was not enlarged; in 201, or 20.1 per 
cent, it was enlarged. Eight, or 1 per cent, of the cases with 
negative spleens were positive for malaria both clinically and 
microscopically. In these cases the spleen was tender in 1 case, 
not definitely mentioned in 3 cases, and not enlarged in 4 cases. 
Of the 201 enlarged spleens, 100, or 49.75 per cent, occurred 
in malarial, 67 in miscellaneous, and 34 in typhoid cases. 

A number of the spleens were stated to be enlarged to percus- 
sion or palpable upon deep inspiration only. In order to arrive 
at a practical splenic index for malaria, it is deemed justifiable 
to exclude these cases. They were distributed as follows: 


TABLE IX.—Spleen enlarged to percussion or palpable on deep 
inspiration only. 


Palpable 
Enlarged 
on deep Total. 


Cases. topercus-| - . 
. imspira- 
sion. cr 
UATE SE [Ec See eee Se ee ee ee P NS Ue ar e 7 2 9 
Mincellancouns isos e mane = toe ee 5 re ee ine a ne oe Nee 17 10 27 


Subtracting these cases from the original ones, there remain 
definitely enlarged spleens in malarial cases 91, or 56.1 per cent, 
in miscellaneous cases 40, in typhoid cases 31. If those in ty- 
phoid fever are excluded, the splenic index of malaria would be 
69.5 per cent, or 91 in 131 cases. 

An analysis of the 40 miscellaneous cases with definitely en- 
larged spleens is given in Table X. It indicates that the pres- 


450 The Philippine Journal of Science 1914 


ence of malaria was not excluded satisfactorily from a number 
of them. Eighteen were examined once microscopically; 1, 
twice; and 21 not at all. Considering the foregoing data, it’ 
seems reasonable to believe that had sufficient effort been made 
some of these cases would have been positive. In other words, 
the splenic index of malaria, typhoid fever being excluded, would 


have been greater than 69.5 per cent in this series of cases. 


TABLE IX.—Forty miscellaneous cases with enlarged spleens. 


Negative! 7 euco- 


No. Sao ieate Diagnosis. 
tion 
i 1 13, 800 | Tuberculosis, pulmonary. 
2 1 5,600 | Tuberculosis, pulmonary; pleuritis serofibrinous; helminthiasis. 
3 1 12,000 | Bronchitis, acute; amcebiasis; nephritis, acute; helminthiasis. 
4 1 Mi eee Ee = 2 Scabies; malaria; helminthiasis. 
5 2 5,400 | Malaria (?), amcebiasis, helminthiasis. 
6 if 12, 940 Cystitis, acute; prostatitis, acute; gonorrhcea, chronic; pyelone- 
phritis(?). 
7 1 5,000 | Fever, denrue; nephritis(?); helminthiasis. 
8 1 10,000 | Fever, dengue; monadiasis. 
9 1 8,200 | Fever, dengue; helminthiasis. 
10 1 5,000 | Yaws, tertiary; amebiasis; helminthiasis; wound, infected, tibia. 
ll 1 4,000 | Fever, undetermined. 
12 1 6, 130 Do. 
13 1 5,400 | Helminthiasis; ameebiasis; blepharitis, acute; dermatitis, acute. 
14 1 7,500 | Rheumatism, acute. 
15 1 10,000 | Pneumonia, lobar; helminthiasis, 
16 1 5,000 | Malaria(?); helminthiasis. 
17 1 7,320 | Undetermined. 
18 1 6,000 | Fever, dengue. 
| 19 1 4,400 | Splenomegaly; wound, lacerated, perineum; wound, bilateral, cervix. 
(i eS es 6,000 | Pleuritis, serofibrinous; helminthiasis; monadiasis; conjunctivitis. 
| 3 eee eee 8,000 | Splenomegaly at post mortem, etiology undetermined. 
| 22a See 5,400 | Fever, undetermined; helminthiasis. 
2 (tail ia 5,000 | Fever, dengue; helminthiasis. 
CA he 4,000 | Fever, dengue; splenitis, acute. 
Din ee ees 14,000 | Yaws, tertiary; helminthiasis. 
24 ea 16,000 ; Gastritis, acute. 
27 |e seen 6.000 | Pyelonephritis; monadiasis; helminthiasis. 
ji #28 [scene oe 5,000 | Nephritis, chronic. 
OO oe ee 10,000 | Ankylosis, monadiasis; helminthiasis. 
Ul eee 4,000 | Fever, dengue; syphilis, tertiary; gonorrhea, acute. 
SU ieee eee 6,000 | Fever, undetermined.’ 
AN IRAE ee eae 10,800 | Ankylosis. 
BS issn ae 9,035 | Undetermined. 
34.4... eae 12,700 | Tuberculosis, pulmonary; nephritis, acute. 
61 ee 4,400 | Fever, dengue; helminthiasis. 
BOT Ss crew eS 8,790 | Nephritis, acute; amcebiasis; helminthiasis. 
| BT eee 7,000 | Pneumonia, lobar; peritonitis, acute; nephritis, acute. 
BS See 5,800 | Nephritis, acute; helminthiasis. 
| 3} Pe Ae ee 8,000 | Dysentery, bacillary; colitis, chronic; acne vulgaris. 
| Cy eee ee 5,000 | Pleuritis, acute; tuberculosis, pulmonary, incipient; helminthiasis. 


IX, B, 5 Willets: Malaria in General Hospital, Manila 451 


Approaching the subject of splenic index from another point 
of view, one may consider what percentage of cases of clinical 
malaria with definitely enlarged spleens was positive micro- 
scopically. Two hundred sixty-six of the 348 cases diagnosed 
as malaria had enlarged spleens. Of the 266 cases, 2 were not 
examined in the laboratory. Of the remaining 264, 225, or 85.7 
per cent, were positive and 39 negative microscopically. 

Of the 39 cases which were microscopically negative, 14 were 
given no quinine in the hospital excepting that contained in iron, 
quinine, and strychnine to 2 of them. Six of these were exam- 
ined once; 5, twice; and 3, three times. Sixteen cases had been 
taking quinine prior to the examination. Thirteen of these were 
examined once; 3, twice. One case had been positive in the lab- 
oratory during a previous admission. He was examined once 
microscopically. Five of the remaining 8 cases were examined 
once; 1, twice; and 2, four times. It thus appears that of the 
39 cases under discussion 24 were examined once; 9, twice; 3, 
three times; and 2, four times. 

The factors to be considered in interpreting these negative 
results are enumerated elsewhere in this report. A number of 
the cases came from districts known to be malarial, and positive 
laboratory findings would undoubtedly have been obtained had 
treatment been delayed; that is, more than 85.7 per cent of the 
cases with enlarged spleens and positive clinically for malaria 
would have been positive microscopically. 

Musgrave, Walker, and others? found malarial organisms in 
the blood of 105, or 41.01 per cent, of 256 individuals with 
enlarged spleens and enlarged spleens in 105, or 31.91 per cent, 
of 329 cases which were positive microscopically for malaria. 
In comparing their results with mine it is perhaps important 
to note that they were dealing chiefly with unselected persons 
whereas my cases were hospital patients. 


SUMMARY 


1. A diagnosis of malaria was rendered in 348, or 5.17 per 
cent, of 6,732 patients; in 10.6, 2.09, and 0.39 per cent of medical, 
obstetrical, and surgical cases, respectively. 

2. Tertain, zstivo-autumnal, and quartan parasites were 
present. 

38. Manila, Mindoro, and Tarlac and Laguna Provinces ap- 
peared to be the chief sources of infection. Many of the in- 


* This Journal, Sec. B (1914), 9, 137. 


452 The Philippine Journal of Science 


fections apparently contracted in Manila probably occurred 
elsewhere. 

4. The majority of the infections occurred among adult male 
Filipinos. 

5. Malarial organisms were found in 85.1 per cent of 341 cases 
examined microscopically. Failure to find them in a higher per- 
centage was probably due to (a) rendering a positive clinical 
diagnosis upon scanty laboratory work; (b) absence of organ- 
isms from the peripheral circulation; (c) errors in the labor- 
atory; and (d) errors in the clinical diagnosis. 

6. Malarial lesions were found in 5.6 per cent of autopsies per- 
formed upon individuals who died at the Philippine General 
Hospital. A correct clinical diagnosis was rendered in 73.7 per 
cent of the cases. Organisms were found microscopically ante 
mortem in 13, or 86.7 per cent, of 15 cases examined. In the 2 
cases which were negative, chronic lesions were present and 
death was not due to malaria. 

7. In miscellaneous laboratory work upon 185 cases which were 
admitted to the hospital because of malaria, and which were pos- 
itive both clinically and microscopically, special attention was 
given to the leucocyte counts. In 1381 of the cases a leucopenia 
was present in 47.3 per cent; a normal count, in 42.7 per cent; 
and a leucocytosis, in 10.0 per cent. Some of the normal and 
abnormally high counts may be explained by the supposition that 
the blood was taken during sporulation in light or chronic cases 
and between paroxysms in heavy acute cases. It is believed, but 
not proved, that intestinal parasitism and disorders of the genito- 
urinary tract are responsible for some, at least, of the normal and 
abnormally high counts. 

8. Typhoid fever being excluded, the malarial splenic index 
was 69.5 among the first 1,000 thoroughly examined medical cases 
admitted to the hospital during the year. Malaria was not sat- 
isfactorily excluded from a number of the cases with’ enlarged 
spleens. Typhoid fever likewise being excluded, malarial or- 
ganisms were found in 85.7 per cent of 264 cases with enlarged 
spleens examined microscopically. Organisms could have been 
found in a number of the negative cases had treatment been 
delayed. 


THE CHIEF INTESTINAL LESIONS ENCOUNTERED IN ONE 
THOUSAND CONSECUTIVE AUTOPSIES IN MANILA 


By B. C. CROWELL 


(From the Biological Laboratory, Bureau of Science, and the Department 
of Pathology, College of Medicine and Surgery, University 
of the Philippines) 


One thousand consecutive autopsies, performed during the 
period of eighteen months from August, 1912, to February, 
1914, have been reviewed to determine the incidence and char- 
acter of the chief intestinal lesions. Especial attention has been 
given to the incidence of intestinal tuberculosis, typhoid fever, 
and ameebic and bacillary colitis. 

The cases were derived from a large general hospital, a hos- 
pital for contagious diseases, and the medicolegal service of the 
city of Manila, so that all classes of cases were included. The 
overwhelming majority of the cases were Filipinos, although 
some other oriental races and some Caucasians were included. 
Two epidemic diseases, bubonic plague and Asiatic cholera, fur- 
-nished 149 of the 1,000 cases, while many more were subjected 
to examination in order to prove or eliminate cholera. These 
latter cases furnished many examples of intestinal disorders, 
as the public-health authorities during the cholera epidemic nat- 
urally selected cases for autopsy diagnosis which gave a history 
either of sudden death or of diarrhceal disease. 

The incidence of parasitic metazoa has not been especially 
investigated in this series; their incidence in 500 autopsies al- 
ready has been reported. Other than numerically the cholera 
cases will not be referred to in this report, as they furnish the 
subject of another paper.? - 


TABLE I.—Incidence of intestinal diseases in 1,000 consecutive autopsies in 


Manila. 

Asiatic cholera 92 
Intestinal tuberculosis 56 
Typhoid fever 39 
Entameebic colitis 31 
Bacillary colitis 25 
Duodenal ulcer 9 
Noninfectious or unclassified inflammatory lesions of 

intestines 57 


In 17 cases two or more of the above lesions were coexistent 
in the same case. For example, 7 of the cases of tuberculosis 


* Crowell and Hammack, This Journal, Sec. B (1918), 8, 157. 
* Crowell, B. C., ibid. (1914), 9, 361. 
453 


454 The Philippine Journal of Science 1914 


died of cholera; bacillary colitis had supervened upon an old 
case of entamebic colitis, The anatomical lesions associated 
with these intestinal diseases have been so characteristic of the 
diseases in which they occurred that their simple notation al- 
most suffices to explain their importance. 


INTESTINAL TUBERCULOSIS 


In view of the well-known prevalence of tuberculosis in the 
Philippine Islands, it was to be expected that tuberculous in- 
testinal lesions would stand first numerically. The latest report 
of the Director of Health shows that during the fiscal year 1913 
tuberculosis was responsible for 17.9 per cent of the deaths of 
the residents in the city of Manila. Our work shows that 5.6 
per cent (56 cases in all) of all cases coming to autopsy had 
intestinal tuberculosis. 


TABLE II.—Causes of death in 56 cases of intestinal tuberculosis. 


Generalized tuberculosis 35 
Perforation of tuberculous ulcer 3 
Intercurrent diseases: 
Asiatic cholera 
Bacillary dysentery 
Typhoid 
Malaria 
Leprosy 
Gastric cancer 
Fatty heart 
Pregnancy 
Postpartum sepsis 
Postoperative peritonitis 
Cholangitis 
Chronic nephritis 


ee 


In a majority of these cases tuberculosis was widespread 
throughout the body, and 35 patients died of generalized tuber- 
culosis. Three cases of perforation of tuberculous ulcers 
occurred, 1 at a point 30 centimeters above the ileocecal valve, 
1 in the cecum, and 1 in the rectum. The other cases died of 
intercurrent diseases. From the standpoint of clinical diag- 
nosis, it would seem important to remember that an extensive 
tuberculous colitis may closely simulate an entameebic colitis, 
and several such cases have been encountered. 

The cases dying of intercurrent diseases require no comment. 
In general, it may be said that tuberculosis as it attacks Filipi- 
nos is very frequently extremely widespread throughout the sys- 
tem and in the intestine the lesions extend throughout a large 
part of its length. 


IX; B, 5 Crowell: Intestinal Lesions | 455 


TYPHOID FEVER 


The frequence of entameebic and bacillary colitis in Manila 
is often referred to, while but little emphasis has been placed 
on the prevalence of typhoid fever. In our autopsy experience, 
it is found more frequently than either entamcebic or bacillary 
dysentery. It will never be known whether this increased sta- 
tistical frequence of typhoid fever is actual or is due to improved 
methods of diagnosis and closer investigation. Thirty-seven of 
the 39 cases occurred in Filipinos, 1 in a Chinese, and 1 in a Japa- 
nese. No typhoid in Caucasians was encountered in this series. 

The following table excellently exemplifies the usual causes of 
death in typhoid fever cases and their relative frequence. In 
22 of 39 cases (56.4 per cent) typhoid lesions were present in 
the colon as well as in the ileum. 


TABLE IlI.—Causes of death in 39 cases of typhoid fever. 


Intestinal perforation 12 
Intestinal hemorrhage 5 
Suppurative nephritis (pyzmic) 2 
Lobar pneumonia 2 
Perforation of gall bladder 1 
Toxemia 17 


Those cases tabulated as dying of toxzemia include those cases 
in which there were no severe gross anatomical lesions explan- 
atory of death other than the intestinal lesions, bronchopneu- 
monia, degeneration of the heart muscle, or other evidences of 
severe toxemia. Thirty per cent died of intestinal perforation 
and 12 per cent died of hemorrhage. 


ENTAMGBIC COLITIS 
TasLe 1V.—Causes of death in 31 cases of entamebic colitis. 


' Liver abscess 9 
Acute peritonitis: 
Perforation * 
No perforation 
Toxzmia 
Intercurrent diseases: 
Lobar pneumonia 
Tuberculosis 
Perforation of duodenal ulcer 
Accident 
Sarcoma 
Cirrhosis of liver 
Pulmonary abscess 
Arteriosclerosis 
Beriberi 
Postpartum sepsis 
® One of these had also a liver abscess. 


Co & po 


to BC) 


456 The Philippine Journal of Science 1914 


The causes of death in entameebic dysentery cases are summed 
up by Strong * as follows: 

Death may occur in ameebic dysentery from the gravity of the intestinal 
lesions; from exhaustion in protracted cases; from severe complications, 
particularly such as peritonitis due to the perforation of an ulcer in the 
large intestine or appendix or an abscess of the liver or lung; from a 
terminal infection sometimes entering through the ulcerations in the large 
bowel; from intercurrent disease, and from severe intestinal hemorrhage. 

All of these conditions, except the hemorrhage, have been 
encountered in our series. 

The liver abscesses here referred to are entamcebic abscesses 
secondary to entameebic colitis, and do not include ordinary 
pyzmic abscesses or those secondary to suppurative cholangitis; 
a number of these cases have been encountered in the present 
series of 1,000 autopsies. 

Liver abscesses occurred in 9 (29 per cent) of our cases of 
entameebic colitis. This represents, it must be remembered, 
the incidence in fatal cases of entameebic colitis, and is no in- 
dication of its frequence clinically. Three of these cases were 
in Americans and 6 in Filipinos, all being males. Five of the 
cases of liver abscess had operative drainage of the abscesses. 
In all of the cases of liver abscess entameebic colitis was present, 
but there has been established no relation between the severity of 
the intestinal lesions and the occurrence of the liver abscesses. 
It is noteworthy that in several cases of liver abscess with ex- 
tensive ulcerative colitis the patients denied any history of dysen- 
teric symptoms. In 4 of the cases there was a single abscess, 
while in the other 5 cases there were multiple abscesses. The 
right lobe was the most frequent site of the abscess, but in some 
cases the left lobe also was involved and in 1 case the Spigelian 
lobe was entirely destroyed. In two cases the abscess had per- 
forated the diaphragm, but in both the destructive process was 
confined to the diaphragmatic surface of the lung by the presence 
of fibrinous adhesions. Four other cases presented a right-sided 
fibrinous pleurisy over the diaphragmatic surface and in some 
cases fibrous adhesions were also present. One of the cases with 
liver abscess died as the result of an acute peritonitis from an 
entameebic ulcer of the vermiform appendix, and in 1 case the 
gall bladder was filled with pus similar to that found in the 
liver abscess. 

The 5 cases referred to in the table as having acute peritonitis 
do not include the cases of liver abscess. In 2 of the cases there 
was actual perforation of the intestine at the site of ulceration, 


° Pub, P. I. Bur. Gov. Labs. (1905), 5, No. 32, 5. 


IX, B, 5 Crowell: Intestinal Lesions 457 


while in the other 3 cases the infection of the peritoneum is sup- 
posed to have entered through the thinned wall of the intestine 
at the site of ulceration. Of the 2 perforations, 1 occurred 
in the descending colon near the sigmoid flexure and 1 in the 
vermiform appendix. The latter case also had liver abscesses. 

The 3 cases referred to in the table as dying of toxemia ap- 
parently died as the result of the severity of the intestinal lesions. 


BACILLARY COLITIS 
TABLE V.—Causes of death in 25 cases of bacillary colitis. 


Toxemia 13 
Complications: 
Acute peritonitis 3 
Abortion 2 
Postpartum sepsis 1 


Intercurrent diseases: 


Tuberculosis 2 
Malaria 2 
Noma il 
Leprosy i 


Fourteen of these cases occurred in children under 7 years 
of age. In 12 of the cases there was involvement of the lower 
portion of the ileum, this involvement varying from a hypere- 
mia to marked diphtheritic inflammation. 

The cases referred to as dying of toxzemia are those in which 
no essential lesions were found outside of the intestine, save 
degenerative lesions, and 6 of those cases had a bronchopneu- 
monia. Stated roughly it may be said that bronchopneumonia 
is present in from one-third to one-half of the cases of bacillary 
colitis in this series, dying as the result of the severity of the 
intestinal lesions. In none of the cases presenting an acute peri- 
tonitis was this lesion an extensive one, and it apparently rep- 
resented only a terminal infection which had passed through 
the diseased colon. 

The occurrence of 3 cases associated with pregnancy indicates 
the danger of bacillary colitis occurring during pregnancy and 
that the two conditions may influence each other unfavorably. 

In 1 case intestinal tuberculosis coexisted with the bacillary 
colitis, and in 1 case entameebic and bacillary colitis were both 
present. It is possible that more careful investigation would 
enable one to recognize a superadded bacillary infection in a 
greater number of the cases of entamebic colitis, whereas it 
is rather difficult of detection in routine examinations of a large 
number of bodies, on account of both the anatomical and bac- 
teriological difficulties in diagnosis. 


458 The Philippine Journal of Science 1914 


It may be stated here that the etiologic agent was n6t isolated 
in all of the cases of either entameebic or bacillary colitis here 
recorded. In the majority of the cases either the entamceba 
or the dysentery bacillus was isolated, but in the remainder 
the diagnosis has been based on the gross and microscopic 
examinations. 

No serious attempt has been made here to differentiate the 
cases of bacillary colitis into types corresponding to types of bac- 
teria, the bacteriological diganosis, when made, having been based 
chiefly on agglutination with a polyvalent antidysenteric serum. 


DUODENAL ULCERS 

A review of the intestinal lesions encountered would not be 
complete without reference to the duodenal ulcers. 

Nine cases of duodenal ulcer have been encountered in the 
series, 6 of which had perforated. In no case had the clinical 
diagnosis of duodenal ulcer been made. In 2 cases a diagnosis 
of cholecystitis was made, and in 1 cholecystectomy was per- 
formed. Severe anemia was apparently prominent in 2 cases, 
for in 1 case the diagnosis of pernicious anemia and in another 
that of secondary anzemia had been made. 

Eight of the ulcers were in the first part of the duodenum, 
while one involved the orifice of the common bile duct, so that 
the bile duct emptied into the base of the ulcer. Contrary to 
rule, 7 of them occurred on the posterior wall, while 1 was 
on the anterior wall, and the position of the other was not 
recorded. Three occurred in the third decade of life, 3 in the 
fourth, 2 in the fifth, and 1 in the sixth. There are in the de- 
partment the records and specimens of 2 cases, not included in 
this series, of duodenal ulcers in infants, 1 aged 6 months and 
1 aged 7 months. 

In the same series, 15 peptic ulcers of the stomach have 
occurred. 

NONSPECIFIC OR UNCLASSIFIED INFLAMMATORY LESIONS 
OF THE INTESTINE 

This very important group includes 57 cases, 35 of which 
occurred in infants and 22 in adults. 

Reference to Table VI will show that there was sufficient 
explanation for the intestinal lesions in all but 5 cases in 
adults. These 5 cases were examined for cholera vibrios un- 
successfully, and their etiology has not been explained. The 
majority of the cases in infants were also found negative bac- 
teriologically for cholera, and none of those included in this 
table presented the characteristic anatomical lesions of cholera. 
Nor were any of these cases in infants of the type of a bacillary 


IX, B, 5 Crowell: Intestinal Lesions 459 


colitis, those cases being included in the previous table. The 
majority of the infants were extremely emaciated, and many 
presented a bronchopneumonia; diseases of the skin were fre- 
quent among them. For the most part, they come under the 
head of those cases which in the hospital wards for children 
are usually styled “feeding cases.” These, however, do not in- 
clude all the cases. It is an open question whether these cases 
are accounted for simply by poor or injudicious feeding, by 
actual, exogenous infection, or by the action of the normal flora 
of the intestine under unfavorable conditions. Careful bacte- 
riological examination of the feces before or after death in 
a series of these cases might produce data of academic or scien- 
tific value, but the practical fact remains that careful nursing 
under hygienic conditions is the remedy. 


TABLE VI.—Associated conditions in nonspecific or unclassified inflamma- 
tory lesions of the intestine. 


Enteritis. Colitis. Enterocolitis. 
Adults, 15. Infants, 11. Adults, 4. Infants, 11. Adults, 3. | Infants, 18. 


1Opium 1. Genitouri- 1. Uremia. | 
poison- nary in- i 
ing. fection. 

. Cholan- 2. Genitouri- 2. Perforation | 
} 


to 


gitis nary in- of gall 

etc. fection. bladder 
into duo- | 
denum. 

-Lobar 8. Uremia. Ge (2) 
pneumo- 
nia. 

~-Lobar 4, Extensive | 
pneumo- burns. | 
nia. 

. Chronic 
nephri- 
tis. 

6. Chronic 
nephri- | a | 
tis. | 

7. Chronic } | 

nep hri- | 
tis. | / 

8. Chronic | 
nep hri- 


i) 


~ 


on 


tis. 
9. Sepsis. 
10. Malaria. 
11. Trauma. 
2 @) 
13. (2) | 
“4 ©) | 


16. (2) 


460 The Philippine Journal of Science 


SUMMARY AND CONCLUSIONS 


In a series of 1,000 consecutive autopsies in Manila, performed 
during eighteen months, aside from the incidence of intestinal 
parasites and tumors and the lesions in bubonic plague, intestinal 
lesions have been encountered in 292 cases. In this series Asi- 
atic cholera (on account of an epidemic occurring during this 
period) stood first numerically. Second in importance was in- 
testinal tuberculosis, and attention has been drawn to the possibi- 
lity of the occurrence of dysenteric symptoms in this condition 
and to the perforation of intestinal ulcers in three cases. Ty- 
phoid fever was present more frequently than either entamcebic 
or bacillary colitis, and these typhoid cases showed a high per- 
centage of perforations (30 per.cent) and hemorrhages (12 
per cent), all of the cases being among Orientals. Entameebic 
and bacillary colitis have been encountered with less frequency 
than the preceding diseases, and have presented many of the 
possible complications and sequele. Liver abscesses occurred 
in 29 per cent of the entamcebic cases, and in 2 cases the in- 
testines had perforated. Bacillary colitis was present more fre- 
quently in children than in adults. Nine cases of duodenal 
ulcers were encountered, 6 of which had perforated, and 15 
cases of peptic ulcer of the stomach occurred in the same series. 
Severe anzmia and symptoms referable to the gall bladder were 
prominent in some of the cases of duodenal ulcer. Unclassified, 
probably nonspecific inflammatory lesions of the intestines, es- 
pecially in infants, occupy an important place, and offer a prom- 
ising field for further etiological study. 


REVIEWS 


A Text-book | of | Physiology | for | Medical Students and Physicians | by | 
William H. Howell, Ph. D., M. D., Se. D., Li. D. | professor of 
physiology in the Johns Hopkins University, Baltimore | fifth edition, 
thoroughly revised | Philadelphia and London | W. B. Saunders Com- 
pany | 19138. Cloth, pp. 1020, 306 figures, $4 net. 


Professor Howell’s textbook, since its first appearance, has 
been the best presentation of this subject for the use of medical 
students. It is particularly fortunate that the demand for the 
book has been so great that frequent revisions are possible. The 
physician or student will find in the text the more significant 
advances in this science almost to date. 

R. B. GIBSON. 


A Text-book of | General | Bacteriology | by | Edwin O. Jordan, Ph. D. | 
professor of bacteriology in the University of Chicago | and in Rush 
Medical College | [dash] | fully illustrated | [dash] | third edition, 
thoroughly revised | Philadelphia and London | W. B. Saunders Com- 
pany | 1913 | Cloth, pp. 1-628. 


The third edition of this already well-known and justly popu- 
lar book deserves praise. The author has covered the ground 
thoroughly, and the important material added to the chapters 
' on cholera, typhoid fever, and leprosy brings this edition well 
up-to-date. 

J. A. JOHNSTON. 


The | Microtomist’s Vade-mecum | a handbook of the methods of | micros- 


copic | anatomy | by | Arthur Bolles Lee | seventh edition | Philadel- 
phia | P. Blakiston’s Son & Co. | 1012 Walnut Street | 1913 | Cloth, 
pp. i— x + 1-526. Price, $4 net. 


Lee’s Microtomist’s Vade-mecum probably needs an introduc- 
tion to but few laboratory workers in English-speaking coun- 
tries. The first edition appeared in 1885 and has been kept up 
to date by frequently revised editions. The seventh edition prob- 
ably includes more accurate information on histological technique 
than any other book in English, giving laboratory methods in 
detail, and making them readily available by a comprehensive 
index. The references to original articles is not one of the least 
valuable parts of the book. 

Be C2 C: 


129558 6 461 


462 The Philippine Journal of Science 1914 


Disease | and Its Causes | by | W. T. Councilman, A. M., M. D., LL. D. | 
professor of pathology, Harvard University | [seal] | New York | 
Henry Holt and Company | London | Williams and Norgate | Cloth, 
pp. i-viili+1-254. 22 text figures. Price, $0.50 net. 


Disease and Its Causes is an example of the exposition of a 
subject for the laity by one of the foremost authorities on 
that subject. In this little volume, No. 68 of the Home Univer- 
sity Library series, Doctor Councilman has very clearly and 
very attractively portrayed disease as “life under conditions 
which differ from the usual.” The development of the science 
of medicine has been forcefully presented, and the causes of 
disease have been classified and displayed as factors producing 
reactions on the part of the body which constitute the symptoms 
of disease. The laws of nature as exemplified in inheritance and 
the normal growth of the body are presented, and their influence 
is shown to be paramount. Every part of the book contains 
something of value, and the book as a whole is one which the 
laity can read profitably without being under the necessity of 
mastering many technical details to understand disease and its 
causes. As an author is mirrored in his writings, this book 
also furnishes the medical reader with a great authority’s con- 
ception of the subject, which might not be gained by a perusal 
of some of his more strictly technical monographs. 


By Coe: 


A | Text-book | of | Histology | arranged upon an embryological basis | by | 
Dr. Frederic T. Lewis | assistant professor of embryology at the 
Harvard Medical School | and | Dr. Philipp Stohr | formerly professor 
of anatomy at the University of Wiirzburg | second edition, with 495 
illustrations | being the seventh American edition of Stéhr’s histology | 
from the fifteenth German edition, edited by Dr. O. Schultze | 
Philadelphia | P. Blakiston’s Son & Co, | 1012 Walnut Street | 1913 | 
Cloth, pp. i—ix+1—539. Price, $3 net. 


The seventh American edition of Stohr’s classical Lehrbuch 
der Histoligie und der mikroskopischen Anatomie des Menschen 
has been prepared by Lewis and is published as of joint author- 
ship. The previous editions of this work have been recognized 
as forming a standard in histology. This edition presents cer- 
tain modifications in the method of presentation of the subject 
matter, and still retains much of the original text and the major- 
ity of its figures. This edition is even an improvement on its 
predecessors. 

Bo) OP: 


IX, B, 5 Reviews 463 


Malaria | etiology, pathology, diagnosis, | prophylaxis, and treatment | by | 
Graham E. Henson, M. D. | member American Medical Associa- 
tion, Florida Medical Association, Southern | Medical Associa- 
tion, American Society of Tropical Medicine, Medical | Reserve 
Corps, United States Army (non-active list) | with an introduction 
by | Charles C. Bass, M. D. | professor of experimental medicine, 
medical department | Tulane University, New Orleans | twenty- 
seven illustrations | St. Louis | C. V. Mosby Company | 1913 | Cloth, 
pp. 1-190. Price, $2.50. 


As the author states in his preface, one of the chief aims of 
the book is to make a presentation of the subject especially 
useful to the general practitioner in malarious regions. In this 
he has succeeded. A large proportion of the work is given to 
the subjects of the pathology, diagnosis, prophylaxis, and treat- 
ment of the disease. The illustrations are reproduced from 
photographs, microphotographs, and drawings. 

M. A. B. 


Tropical Diseases | a manual of the diseases | of warm climates | by | Sir 
Patrick Manson | G. C. M. G., M. D., LL. D. (Aberd.) | fellow of the 
Royal College of Physicians, London; [etc., 7 lines] | with 12 colour 
and 4 black-and-white plates | and 239 figures in the text | fifth edition, 
revised throughout and | enlarged | New York | William Wood and 
Company | MDCCCCXIV | Cloth, i-xxiv+1-987. Price, $5. 


In this edition of Tropical Diseases much new material is 
added to an already excellent book. The relatively small size 
of the volume, 12 by 18.5 centimeters, makes it convenient for 
one who is unable to carry larger reference books with him. 

M. A. B. 


Marriage and | Genetics | laws of human breeding | and | applied eugenics | 
by | Charles A. L. Reed, M. D.; F. C. S. | fellow of the College of 
Surgeons of America; | member [etc., 3 lines] | The Galton Press, 
Publishers | Cincinnati, Ohio, U. S. A. | Rubber-stamped, Date of 
issue Sep. 10, 1918. Cloth, pp. 1-188. Price, $1. 


The book springs from a desire in some measure to overcome 
the ignorance which, in too many instances, keeps innocent vic- 
tims from protecting themselves and their offspring from disease 
and degeneracy. The problem is vital. It is first an individual, 
then arace problem. The question, “what of me and my family,” 
must be asked before that of ‘‘what of my neighbors and their 
families.” Both must be asked, but this is the necessary order. 

The problem deals with the deepest human sentiment and the 
profoundest welfare of society now as well as in the future. 
The book attempts to present as simply as possible the funda- 
mental laws of race perpetuation, considering causes and effects 


464 The Philippine Journal of Science 1914 


in relation thereto and indicating some of the measures available 
to society to repress or to eliminate practices and hereditary 
strains that tend to degeneracy, and to foster as far as possible 
those that tend to human betterment. The Introduction, and 
the two divisions entitled The Race Poisons, and Applied Euge- 
nics, may be easily and profitably read by any intelligent person, 
and most persons can follow the other division called General 
Laws of Genetics even though they are not already familiar 
with the laws of Weismann, Haeckel, Galton, or Mendel. In 
fact, a careful study of Reed’s presentation of these laws will 
make a good introduction to them and well repay the reader 
for his effort. 

The message of the book is especially addressed to every pros- 
pective husband and wife. Knowledge is the best preventive of 
disaster. The book should have extensive and thoughtful 
reading. 

C. E. Cox. 


Further Researches | into | induced Cell-reproduction | and Cancer | consisting 
of papers by | H. C. Ross, M. R. C. S. England, L. R. C. P. 
London | J. W. Cropper, M. B., M. Se. Liverpool, and | E. H. Ross, 
M. R. C. S. England, L. R. C. P. London | with illustrations | the 
McFadden | researches | [Rubber-stamped: P. Blakiston’s Son & Co., 
Publishers, | 1012 Walnut St., Philadelphia] | London | John Mur- 
ray, Albemarle Street, W. | September 1911 | Cloth, pp. 1-63. 
Price, $1 net. 

Muscle Spasm and Degeneration | in intrathoracic inflammations | their 
importance as diagnostic aids and their influence in producing and > 
altering | the well established physical signs, also a consideration 
of their | part in the causation of changes in the bony thorax | 
and | light touch palpation | the possibility and practicability of 
delimiting normal organs and | diagnosticating diseased conditions 
within the chest and | abdomen by very light touch | by | Francis 
Marion Pottenger, A. M., M. D., LL. D. | medical director of the 
Pottenger Sanatorium for Diseases of the Lungs and Throat, | 
Monrovia, California | sixteen illustrations | St. Louis | C. V. Mosby 
Company | 1912 | Cloth, pp. 1-105. 


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Vou. IX, SEc. B, No. 6 NOVEMBER, 1914 
THE PHILIPPINE 


JOURNAL OF SCIENCE 


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GENERAL EDITOR 


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THE PHILIPPINE 


J OURNAL OF SCIENC 


B. TROPICAL MEDICINE 


VoL. IX NOVEMBER, 1914 No. 6 


THE GERMICIDAL POWER OF GLYCERIN ON VARIOUS MICRO- 
ORGANISMS UNDER VARIOUS CONDITIONS 


By E. H. RUEDIGER 
(From the Biological Laboratory, Bureau of Science, Maniia, P. I.) 


Although glycerin is extensively used, especially as a preser- 
vative in vaccine virus, very little has been published regarding 
its germicidal properties. Rosenau‘* seems to be the only one 
who has made any extensive study of these properties. The use 
of glycerin as a germ-destroying agent in the preparation of bac- 
terial vaccine introduced by Row? has aroused new interest in 
the subject. The tests reported here were made in order to 
obtain accurate information as to the extent that glycerin may 
be relied upon to sterilize bacterial vaccines. 


THE GERMICIDAL POWER OF GLYCERIN IN PHYSIOLOGIC SALT 
SOLUTION AT ROOM TEMPERATURE 


TEST 1 


Eight sets of test tubes were prepared, and the sets were num- 
bered 1, 2, 3, 4, 5, 6, 7, and 8. Each set contained four tubes, 
a, b,c, and d. Into tube a were put 2 cubic centimeters of salt 
solution (9 grams per liter) ; into tube b were put 2 cubic centi- 
meters of 12.5 per cent solution of chemically pure glycerin; into 
tube c were put 2 cubic centimeters of 25 per cent. solution of 
glycerin; and tube d received 2 cubic centimeters of 50 per cent 
solution of glycerin. The tubes with their contents were ster- 
ilized in the autoclave. After sterilization the 8 sets of tubes 
were inoculated with the typhoid bacillus, Staphylococcus albus, 


* Bull. Hyg. Lab. U. S. Pol. Hith. & Mar.-Hosp. Serv., Wash. (1913), 
No. 16. 
2 Journ. Trop. Med. (1913), 16, 293. 
129562 465 


466 The Philippine Journal of Science 1914 


Staphylococcus aureus, the bacillus of anthrax, the bacillus of 
plague, the spirillum of cholera, the bacillus of diphtheria, and 
with the bacillus of glanders, respectively. Each tube received 
about one billion organisms. 

For a period of fifteen days a 2-millimeter loopful of the bac- 
terial suspension from each tube was planted on slanted nutrient 
agar, and after from three to five days’ incubation the results 
were recorded (Tables I to VIII, inclusive). The plus sign (+) 
in these tables means that growth of the organisms was obtained 
on the agar, while the zero sign (0) stands for no growth on the 
agar. Y 
This work was done in March, April, and May, 1914, during 
which time the temperature varied from about 28° C. to 35° C. 
No tests were made at incubator temperature, 37° C. 


TABLE I.—The action of glycerin in salt solution at room temperature on 
the typhoid bacillus. 


Days. 
Glycerin. a 
1 2 3 4 | 5 | 6 | 7 8 9 10} 11 | 12 | 18) 14 | 16 
| 0 per cent _--..-. +)t] +} 4+) 4+ +14 t+ ]4+] 4+), +)4+) 4+] +414 
12.5 per cent-____- arene dtaie ece tear esol Gellar cmalle seller Sellen (hh 
25 per cent _____. speliecs It iam «peek |) camels cae |(o stm lt, oe 0 0 0 0 0 0 0 
50 per cent --_--- se} Sr Nae 0 0 0 0 0 0 0 0 0 0 0 0 


TABLE II.—The action of glycerin in salt solution at room temperature on 
Staphylococcus albus. 


| Days. 
Glycerin. ft Se ke eS a Se 
la 2 8 4 5 A 8 9 | 10 ae 18 | 14 | 15 
+ 
12.5 per cent-_---_- + 
25 per cent -._.-- 77 
60 per cent ____-- =F 


TABLE III.—The action of glycerin in salt solution at room temperature on 
Staphylococcus aureus. 


Days. | 
Glycerin. St 
1 2 3 4 5 6 7 8 9 | 10] 11 | 12] 18] 14] 15 


0 per cent -______- a foam or fea Pca eee Poort tse ifr oe pase jp ose 
12.5 per cent____- am jar War Sehr ies [seb eeae hse les eses yb 0 0 
25 per cent___.__. ar i} Sry ar iu 0 0 0 0 0 0 0 0 0 0 0 
60 per cent_-____- aed) 0 0 0 0 0 0 0 0 0 0 0 0 0 
meas TO ee) 


IX, B, 6 Ruediger: Germicidal Power of Glycerin 467 


TABLE 1V.—The action of glycerin in salt solution at room temperature on 
the bacillus of anthrax. 


| | Days. 
Glycerin. _ ; 
| 1 | 2 3 | 4 5 | 6 7 8 9 | 10; 11 | 12) 13] 14) 15 
0 per cent___----- me ae ih 4 a ae a tae epee 
W6perecent.....) +] +) +/+), +) +1 4+) +), t+) 4+, F] +p they 
. 25 per cent_-.---.| + | + | a | ite | + | + | +] 4 | Sef ee tN Se) pe 
50 per cent_._.--- aa ee ne |e ibe al vale cal ea Galion 


TABLE V.—The action of glycerin in salt solution at room temperature on 
the bacillus of plague. 


[ Days. 
Glycerin. aiid Gina eaall = ; - 

1 2 3 5 6 | 7 8 | 9 | 10) 11 12 | 18 peal 

Cee ee ee ee ee ee OO 

0 per cent._-_-_-- + ear ipa +} +|+ 0 0 
12.5 per cent _-____ Ete (ara sce | tring 0 0 0 0 0 0 0 0 0 i 0 0 0 
25 per cent__-___- lt 0 0 0 0 0 0 0 0 0 | 0 0 0 0 0 
60 per cent_____-_ 0 0 | 0 0 0 0 0 0 | 0 0 | 0 0 0 0 0 | 


TABLE VI.—The action of glycerin in salt solution at room temperature on 
the spirillum of cholera. 


Days. 
Glycerin. Ma EAT MIE ERG ELST GRaL ON ClO i la ae 


| 1 2 8 4 5 | 6 7 8 | 9 | 10 | 11] 12) 18 | 14) 15 
0 per cent__--_-_-- ar (tote apg ap dy GE | anwar Ge alos ed Sear Boe Ware | loc mld ay 
12.5 per cent ____- Sell ey ar atl Sel ore se) MS el et a ae ar | Nies SI  iea ce 0 
25 per cent_.___-- + a 0 0 0 0 0 | 0 0 | 0 0 0 0 0 0 
50 per cent______- Ge io) PO. On (0 Om ino 0 | 0 | OOM nO Onl on tO 


TaBLE VII.—The action of glycerin in salt solution at room temperature on 
the bacillus of diphtheria. 


: ; Days. 
Glycerin. se] e]*] jele]s| selma — 


| 1 


oo 
~ 
o 
> 


0 per cent_._-_._- acnets 
12.5 per cent __... RE | se 
+ 
+ 


468 The Philippine Journal of Science 1914 


TABLE VIII.—The action of glycerin in salt solution at room temperature on 
the bacillus of glanders. 


| | Days. 

| Glycerin 

| 1 2 | 3 4 By | 6 7 8 9 | 10} il 12 | 138 | 14 | 15 

' Oper cent_______- ao ee Re RS seep Rep ep ep Re Se Se | 
| 12.5 per cent _____ +/+]a]e}4}a]4]4]4}4]4]4}4)4]4 

| 25 per cent... | + | | so) | 0p Ni Ore Ono | 0 | OF | 0: | 0. | som aagn 

| 50 per cent_______ 0 0 0 | 0 | 0 0 0 | 0 | 0 | 0 | 0 0 | 0 0 0 


It will be seen from the results recorded in Tables I to VIII, 
inclusive, that the germicidal power of glycerin varies with dif- 
ferent microérganisms. In 50 per cent of glycerin in physiologic 
salt solution the typhoid bacillus lived three days; Staphylococcus 
albus and aureus lived one day; the plague bacillus, the spirillum 
of cholera, and the bacillus of glanders were dead before the end 
of twenty-four hours; diphtheria bacilli lived two days; while the 
bacillus of anthrax was alive at the end of fifteen days. Twenty- 
five per cent glycerin was sufficiently active on nonspore-forming 
organisms to receive mention. In 25 per cent glycerin the ty- 
phoid bacillus lived eight days; Staphylococcus albus, Staphy- 
lococcus aureus, and the glanders bacillus lived three days; the 
diphtheria bacillus lived four days; the spirillum of cholera, two 
days; while the plague bacillus was dead on the second day. 


TEST 2 


Because of the great importance of the plague bacillus 
in plague-infected countries, the germicidal power of glycerin 
in 12.5 per cent, in 25 per cent, and in 50 per cent dilutions in 
physiologic salt solution, at room temperature, was determined 
on 4 different strains of plague bacilli. The results are re- 
corded in Tables IX, X, XI, and XII. 


TABLE [X.—The action of glycerin in salt solution at room temperature on 
plague bacillus No. 2. 


Vinee Seer ere a 


Glycerin. 
r}2e/s|/4i6j]6]71]8|9 | 10/41 | 12] 18] 14] 05 
O percent _______ (i ic a aa cae We eee ciel ate ace llanieyy 2 
12.5 percent ---_- “fe | See ten O) (Ot Oy. (Oo), 0) ia ea O we te Os ano OM OO! 
85 per cent..___- atheghhooel: ofa | o-}oc! to) phone serine meamiany 
50 per cent_______ 0 | 0 0 0 | 0 0 0 0 0 0 0 0 | 0 0 | 0 
1 


x) B, 6 Ruediger: Germicidal Power of Glycerin 469 


TABLE X.—The action of glycerin in salt solution at room temperature on 
plague bacillus No. 3. 


Days. 

Glycerin. aT =o 
1 2 8 4 5 6 7 8 9 | 10 11112 )18 ) 14) 15 

DEES ane See Ba eee ea | = ene 
0 per cent_..._____ ae Srp se ar ar Wesel Sp Se A sae date tard eel) Se crt 
12.5 per cent _-___- Set arp dl Oy ae ae) 0 0 0 0 0 0 0 0 |.0 0 
25 per cent _____-- ae Hoare ed 0 0 0 0 0 0 0 0 0 0 0 0 
50 per cent____..__ 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 


TABLE XI.—The action of glycerin in salt solution at room temperature on 
plague bacillus No. 4. 


a ‘nga Days. bees ye aie 


Glycerin. 


1 2 8 4 5 

| Be a ss a 

0 per cent___--_-- ee Wear ose ase ge oe tess ise i ae ae Tae ae) pall Sim aC) 

‘12.5 per cent _-___ Se th ae dl sp tw 0 0 0 0 0 0 0 0 0 0 0 | 

25 per cent____--_- ae ow) 0 0 0 0 0 0 0 0 0 0 0 0 oO | 
50 per cent______- 0 0 0 0 0 0 0 tt) 0 0 0 0 0 0 0 


TABLE XII.—The action of glycerin in salt solution at room temperature on 
plague bacillus No. 5. 


a 


| Days. 
Glycerin. ] Ree Se 

1 2 3 4 5 6 | a 8 9 | 10] 11] 12 | 18 | 14] 15 
0 per cent-____-_-- Sel aR Se Sr) ae lar ese Se I See ae 4 an | ary} Ww 0 
12.5 per cent _____ ap ll ose 55 0 0 0 0 0 0 10 0 0 0 0 0 
25 per cent-_ .__._- + 0 0 0 0 0 0 0 0 0 0 0 0 0 0 
50 per cent_______ 0 0 0 0 0 0'| 0 0 0 0 0 0 (1) 0 0 


The results in Tables IX to XII show that the action of gly- 
cerin on different strains of plague bacilli varies but slightly; 
the differences are perhaps not greater than those obtained by 
suspending the germs in plain distilled water or in plain physio- 
logic salt solution. Of the controls (those kept in physiologic 
salt solution), 2 strains were found dead on the fourteenth day 
and the other. 2 strains were dead on the fifteenth day. 

In 12.5 per cent glycerin 3 strains gave negative results on 
and after the fourth day, while 1 strain gave a negative result 
on the third day, and 3 colonies developed on the agar inoculated 
on the fourth day. 

In 25 per cent glycerin 8 strains were killed in two days and 1 
strain was killed in three days. 


A70 The Philippine Journal of Science 1914 


Inoculations made from suspension in 50 per cent glycerin 
twenty-four hours after the suspensions were made gave neg- 
ative results. 

TEST 3 

Hight sets of test tubes were prepared as in test 1, and the 
mixtures were placed in the refrigerator and kept at a tempera- 
ture of about 15° C. Each day the tubes were removed from 
the refrigerator to inoculate a tube of agar with one 2-millimeter 
loopful of bacterial suspension from each tube. The time re- 
quired for inoculation was short and should not have greatly 
affected the temperature of the suspension or the action of the 
glycerin on the bacteria. Tubes of nutrient agar were inoculated 
daily for a period of fifteen days; the results are recorded in 
Tables XIII to XX, inclusive. 


TABLE XIII.—The action of glycerin in salt solution at a temperature of 
15° C. on the typhoid bacillus. 


Days. 
Glycerin. ee Sel Meee. Se CEE ESE, Seen RMN, | Ae WN A oe Ds te iit 
| 15 


| 

| 

| rl ek Juliet ioe Saltash calles » | 10 11 | 12 18 | 14 

| 

{mens ae  E SES [PS a fle aw | fe Ya a a ed ed et 
0 per cent ----___- eum linet | oe a Wee | (ea i Oe ia enh a A etme sae Sp 
12.5 per cent -__.. | +} 4 | tho] eptia> +} a+] ti} 4e] 4] 444+ 
25 per cent _____- p+ itp t] t+] 4] 4 Pe fem coke dg AOS ge 

! 50 percent _-..__- +i ti +] +] a+] + ee +)epe|ol4 0 | 0 

| | : ie | : ih salle sat | i | i" lk | 1 Bien 


TABLE XIV.—The action of glycerin in salt solution at a temperature of 
15° C. on Staphylococcus albus. 


Ja : ie: GREP eM unD rare ae ps, 
| Days. 

| Glycerin. 5 era 1 Tae ye al ee is es ee? 

| 1/2}]s8|4 se] 7] 8/9 10} 11] 12 | 13 | 14] 15 
| (Opereent=.-.- 5 Speier tom eee df) amt ar | =F | ar dba lose Weer |b eee meats If op 
| 12.6 per cent ____- +) 4a] +] 4]4+]4]4 | +/+} aya] +] +] + 4+ 
| 25 percent ---.-- +/+} +] 4] 4+ y+ | im +i +) 4+] 4+] 4) +)+ 4+ 
| ipo secant a. half ag) balla tt Noes | eta at “gh at gid 0 oul 


TABLE XV.—The action of glycerin in salt solution at a temperature of 
15° C. on Staphylococcus aureus. 


D je Syuies a i} ss 10} 1 | a2 | 15 
F Ty Pale ea ST ea nik ac a 

| Oper cent_.-.-_.. se Neste in| eka hata gh oat ieestcet ct omits |) cma tariler (lar sar |) ae 
12.5 per cent ____- Srlvenm cam ber Wl, tap | ay |)ar | Riel ies aii rural (vcore tN loners ir meyl} eed forcsne 
25 per cent_-_____- AE REE Raia by ee | Se ore same) aa bie leper ih ae |) seb se a ae 

; 50 per cent______- =f | moeliiae |r | 3 | aF | }- | amp tear Lar SP i ar oil ae | cts uz 

— a as ers tn eoetineon 


Eo Eu. ‘dh wel Hd Gh. a | 
Glycerin. ae 2 ape aso ——— ieee Foe 
lake | 


— 


Oe B.. 6 Ruediger: Germicidal Power of Glycerin 471 


TABLE XVI.—The action of glycerin in salt solution at a temperature of 
15° C. on the bacillus of anthrax. 


Days. 
Glycerin. — = 


ple | 3 | 4 | 5 | 6|/ 7/8 | 9 | 10] | 12] 18 | 14] 15 

0 per cent -------- + | +/+ ft] +)+ y+} + y+} 4+ +74 p+ yryt 

12.5 per cent ____- ap Se ba ee ee ad a em ee tl eet ge eae Horie 

| 25 per cent ______- Ap Woe i aN Se Sat Set ae eae ih ae ae) et rel Sarlae 

| 50 per cent... +i +] + ]4+ir>4+}4+]4+]4}+]4+ + | cae al ee 
fen ee ne A BE bon ee ee ee ne ee 


TABLE XVII.—The action of glycerin in salt solution at a temperature of 
15° C. on the bacillus of plague. 


Days. : 
Glycerin. Ee Rone eA Se ES a Ee ee | 


1 2 3 4] 5 6 q 8 | 9 | 10] 11 | 12} 13 | 14} 15 
| 0 per cent -._.___- sp for la \) se ae lee ose ae tse (lee See buss |) ae A 
; 12.5 per cent ----- Se qeSp doar sr ae |) Se oiosr tee bese i ee ber WSe te ae ae 
| 25 per cent____-__ se ge diese ee ll apd Seciivae ise iba tse diary ae Negra sae) oe 
| 50 per cent ______- | Be ONS SRA ee EP aR 0 0 0 0 0 0 0 0 0 


TABLE XVIII.—The action of glycerin in salt solution at a temperature of 
15° C. on the spirillum of cholera. 


i Glycerin. - —- ; ep 

SU OCR Oa 5 | 6 7} 8 | 9 | 10 aA 14 | 15 

| oe 

' Oper cent -______- +Pepe [tpt ye Pt pe] tye tees | a ee 

| 12.5 per cent _____ t+htte]rtit]}at+y tit] +t +} 4+i 4+) 0!o 
25 per cent _______ Hele eee se oe tO Oy {Oo at 0) | o. To 
60 per cent ___-__- Saree Ons One Oeil ED. LOFT O: }5 Oh) OPO oO 


TABLE XIX.—The action of glycerin in salt solution at a temperature of 
15° C. on the bacillus of diphtheria. 


Days. 

| Glycerin. 

| r{els ae an 8 | 9 tli 12 | 18 | 14 | 15 

| 0 per cent_______- Beh an | SEW Sart Sar dp Suey ard reset erat Ae etn oa re pea (a | 
Vamppencent See ie eee ere ie (ee) Fe) ee eee oe 

| 25 per cent_______ ar war dae se fioge |] Se de a ern eos med) ae re te Se Wher 

| 50 per cent__.--_- fe ae hose ar ae Wes [se sey Se ae) ay ag 


L 


472 


TABLE XX.—The action of glycerin in salt solution at a temperature of 
15° C. on the bacillus of glanders. 


The Philippine Journal of Science 


| Glycerin. 


The results obtained and recorded in Tables XVIII to XX, 


1914 


Days. 
6 | 7 8 | 9 
ap i} ar |) Se ae 
Sey) Sb i spelt oe 
ae i) Sp Uo ae i) Se 
Sp i) ae | se | ar 


10 | rb 
+] 4 
+] + 
+] + 
+] 0 


inclusive, show that at low temperature the germicidal power 


of glycerin in physiologic salt solution is very slight. All the 
organisms except the spirillum of cholera lived fifteen days in 
In 50 per cent glycerin these organisms 


25 per cent glycerin. 


lived too long to be of any practical value in the preparation of 


bacterial vaccine. 


Eight sets of test tubes were prepared as in test 1, the gly- 


TEST 4 


cerin being diluted with nutrient broth (bouillon) instead of with 
physiologic salt solution. 
oculated with about 1,000,000 bacteria. 
kept at room temperature, and tubes of nutrient agar were in- 
oculated daily with the suspensions of bacteria in the diluted 


glycerin. 
inclusive. 


Each tube of diluted glycerin was in- 


The suspensions were 


The results are recorded in Tables XXI to XXVIII, 


TABLE XXI.—The action of glycerin in bouillon at room temperature on 


| Glycerin. 


the typhoid bacillus. 


© pericent.2/ 615. 
12.5 per cent __._- 
25 per cent_______ 
50 per cent_____- 


TABLE XXII.—The 


t+ + 
++4+4 


o++ + 


(Oot 4+4 


Days. 


action of glycerin in bouillon at room temperature on 


Staphylococcus albus. 


Days. 
Glycerin. ta. FAT ee Waal k al a x cies 

Re Zee ere eee Bie Gas (2, | 8 | 9 10 | 11 | 12; 13] 14) 15 

oe oes eee alk Fj Me 
0 per cent_______- ae a iota ae omtectos Miata ect | 4 & Ce SF ol Pater Ap ate ge nd | eRe 
12.5 per cent ____- SoU CS cre or pe Mercalli miecrib or lberm llcams oreaf om | s. 
| 25 per cent__-_-_- oie cae ite se in i a WW ce cael sats sh 
50 per cent______- Sd Ste ccta a erie Hivtate aaliaectea atct= a] On LO con |e Mange uO a OU a | 


IX, B, 6 Ruediger: Germicidal Power of Glycerin A473 


TABLE XXIII.—The action of glycerin in bouillon at room temperature on 


Staphylococcus aureus. 


Glycerin. 


0 per cent ________ 
12.5 per cent ____- 
25 per cent _______ 
50 per cent _____-- | 


Days. 
| ine | H 
1 2 3 4 5 6 | 7 8 | 9 | 10 | 11 | 12 | 13 14] 15 
i 
ai tal! ons | Sp Mae ad Sora) ae | ote ee a oe eA Seren Mola ar | eau 
Se At eerie] et SN | ae a mete (ae Te a STAN ede tee an ee 
Sa Vier er ine a Pc a Vm oe ee Ce Vr Ue Do a 
Blt tl ey eye et pe ee a a ea 
1 1 


TABLE XXIV.—The action of glycerin in bouillon at room temperature on 


| | 
Glycerin. 


0 per cent _-_-_--_- 
12.5 per cent .___- 


the bacillus of anthrax. 


25 per cent ______- 
60 per cent ___.- tx 


Days. ! 
1 2 3 als | 8 9 | 10 ni 1 13 | 14} 15 | 
ap Sr |i ae oar yar sea Pam Ge dh aro Se aR See ae I ose 
ar il seeil ae War or ava Wasp oteais | ok ate an i] Se eral (ected 
Se a oe  orallaey) om eer aol ae 
Be) eel Grad) Mazel) cee) ae ta Sia See Meret eae ce oo Beal op 


TABLE XXV.—The action of glycerin in bouillon at room temperature on 


the bacillus of plague. 


Glycerin. 
1 
0 per cent ____.___ Sm alk oe ves 
12.5 per cent _____ ap i sew ae il ae 
25 per cent _____.- ee ae aes 
ae) 0 0 


| 50 per cent _______ 


TABLE XXVI.—The 


action of glycerin in bouillon at room temperature on 
the spirillum of cholera. 


Days. \ 

Glycerin. 

Teal a | 4 Bel sGele sso) 9 10 | 11 | 12 13 | 14 15 | 

=" | ii | I ary Zl | | F 
0 per Bsa aly Steel stats | vais Vi iaie en acter che tipestal anche rie ce iret Mest dnote Wi Sie oie 
12/5 percent} +} Ee ee eee eae ee eS 
25 per cent.--.--- Napili 2 ule Gi) Oidie@ il, Orl (Ord Only 0 yo 
50 per cent___.___ OP OO] OO OO] OOO | 0 10 sb-0 | oO 1-0 

} i 


AT4 The Philippine Journal of Science 1914 


TABLE XXVII.—The action of glycerin in bouillon at room temperature 
on the bacillus of diphtheria. 


| 

Glycerin. emiat: ss 

| | 1 2 3 | 5 | 6 7 | | 9 | 10] 11 12 13 | 14 15 
0 per cent____---- ai lle | Ste tant eat | ron! aera eueel eae | SR oe iced Set Se) ap 
12.5 per cent__-_- +] ape pe le ie | Sel) SP) ae eae) Grek se ar} aro) ae 
25 per cent_---.-. +J+]a] 4! a] at+} ate }4!4] 4+! of] oto 
50 per cent__--__- aR SE) se es + | 0 0 | 0 o| 0 0 | 0 | 0 


TABLE XXVIII.—The action of glycerin in bouillon at room temperature on 
the bacillus of glanders. 


Days. 
Glycerin. Fn 
1 2 8 [als a 9 | 10 | 11 12 13 | 14 16 


0 per cent_-__--_- Ge Pier wt mre Gore Geol) orate) Se Se eth or ie) Sp Se 

12.5 per cent__-_- ant crea e|| Pal earl caer ears here bce eer im Hose | so 3 
25 per cent -_-____ ae isenit Se Sarat eee oy ty oj; 0 0; 0 0 0 0 0 
50 per cent__-___- + | sreneeste nit pete || 200 | 0 0 0 0 | 0 0 0 0 0 | 0 | 


From the results recorded in Tables XXI to XXVIII we learn 
that glycerin mixed with bouillon has but feeble germicidal 
power. In 12.5 per cent of glycerin, all the organisms lived 
fifteen days. The typhoid bacillus, Staphylococcus albus, Sta- 
phylococcus aureus, and the anthrax bacillus were alive in 25 
per cent glycerin at the end of fifteen days. In 50 per cent 
glycerin the typhoid bacillus live three days, Staphylococcus 
albus lived four days, Staphylococcus aureus lived five days, the 
bacillus of anthrax lived fifteen days, the bacillus of plague lived 
one day, the spirillum of cholera was found to be dead at the 
end of one day after inoculation, and the bacillus of diphtheria and 
the bacillus of glanders lived six days and four days, respectively. 


TEST 5 


Eight sets of test tubes were prepared as before. Horse serum 
was used as a diluent for the glycerin. The tubes were inoculated 
and were kept at room temperature, and for a period of fifteen 
days a 2-millimeter loopful of bacterial suspension from each tube 
was transferred to a tube of agar. The results are recorded in 
Tables XXIX to XXXVI, inclusive. 


IX, B, 6 Ruediger: Germicidal Power of Glycerin 475 


TABLE XXIX.—The action of glycerin in horse serum at room temperature 
on the typhoid bacillus. 


Days. 
Glycerin. | F 7 7 i 
1 2 3 | 4 ay tak oa ba 8 9 10| 11] 12) 18] 14] 15 | 
0 per cent -_---.-- Siaveeche (che chy ase Ny oe che 41+ anette fe Galan meet oer 
| 12.5 percent _---- +ley+feyef eye fey ele] eye] eys | + 
| 25 per cent ------- Seo Sell api sae ll Se | Seed eel eal Paral) aed ral ee Se  () 
50 per cent _____-- + 0 0 0 0 | 0 0 0 0 0 0 0 0 0 0 


TABLE XXX.—The action of glycerin in horse serum at room temperature on 
Staphylococcus albus. 


{ 

Days | 

Glycerin. a 

rie] s 4/65] 6 z|slo 10; 11 12 | 13 14 | 16 | 

0 per cent ____-__- te orto eet Doce Cerra ee yarn Tee f | sol) ane ian li ue eer iro he eal le | 
12.5 per cent ____- iat Spret saedl me ay pear tier yl oma ene ah Sorrell (get | Pencil) aud Uh font bet 
25 per cent ______. SoU Sa sb ee a) Se a eee! ere See eal Sarath dived ar 
50 per cent _______ qe ose |b ae 0 ae 0 0 0 0 0 0 0 0 0 0 


TABLE XXXI.—The action of glycerin in horse serum at room temperature 
on Staphylococcus aureus. 


| Days. 

( j 

eee Qi ea Sie 

1f2|a [alo z[s [9 [10/1 | a2 [a8 | 14 | 15 

2 cae eae Pa cet iene rh ek 
0 per cent -____._- (ae oreo ay ea ee: 

| 12.5 per cent -__. +Jtle] 4] + A et Es ge ie 
25 per cent _------ Poo ei arabs ear eho pe ee 
50 per cent ______- na ea an cag | 7G ole a 0 0 eae 


TABLE XXXII.—The action of glycerin in horse serum at room temperature 
on the bacillus of anthrax. 


| [ 
| Die iteite perpen leg | 8 | 9 | 10) 11 | 12 | 18 | 14| 15 
A ad re a eas 
| Oper cent_____._. eal es Beebe | Ph [tlt bth be 
| 12.6 per cent __.-_| Boy SE aie ouh Boe! 2) ie Gg ares BN bee | A (reas a Baer ie | 
25 per cent... fos | Bh ae ee Ge Se) ee Ee 9 ay ae ee 
50 per cent __.... +]e i) aye fe] ey tye) 4) 4] + | fo) Be + | 


476 The Philippine Journal of Science 1914 


TABLE XXXIII.—The action of glycerin in horse serum at room temperature 
on the bacillus of plague. 


Days. | 
Glycerin. 
| 15 


1 | 2 3 | 4 5 | 6 7 | 8 9 | 10 | 11} 12 | 13 | 14 
i: Penne SIDS TD Ep) (SE UME CEQ fe (CTs) Ke EG) a SO ee UE ERS MeN oss Dace 
0 per cent _______- cpiulear (lower isan Woon ae ae ope ee Papi Sr pa) 
12.5 per cent _____ ar Sie th sr i) ar io ae lh sr ae i srip ap lise 0 0 0 0 0 
25 per cent ___-___ me il 4 ap {lGr ae i) on + 0 0 0 0 0 0 0 0 
50 per cent -___-__ oF 0 | 0 0 | 0 | 0 | 0 0 | 0 0 | 0 0 0 | 0 | 0 
1 


TABLE XXXIV.—The action of glycerin in horse serum at room temperature 
on the spirillum of cholera. 


0 per cent _____. __ ae 


Hediste eee |b ee real eae bea Bal oy | See ae 
12.5 per cent _.___ spilicabaey re iiipeecr | celle | eM bertaa lie tall aa ae 
25 per cent _______ +) 4/4) 4+), +] 0] 0 1/0 | 0.) 0 | 1o,] 0 | OL ono 
50 per cent _______ OmO) ino oo 0 | 0 ojo 0) 0.) (0 | Opie a 


TABLE XXXV.—The action of glycerin in horse serum at room temperature 
on the bacillus of diphtheria. 


Days. 
Glycerin. aE . Bs ah ! 


0 per cent -___--_- -+ + + Ss “if =f ae ai = ae ae sa ap ll ar = 
12.5 per cent ---_- Si | coll, Tel oy ee Meer an (eae |e Smale amar |i sun (far 
25 per cent _--___- Satin eaee cela OS lea hse Woah ooo ie (he tl i Me olh | 
50 per cent _-_____ aP | Sal Wea | ate leets | 0; 0 | 0] 0 | 0 | 0 | 0 |] 0 | 0 | 0 | 


i 


TABLE XXXVI.—The action of glycerin in horse serum at room temperature 
on the bacillus of glanders. 


Days. 

Glycerin. ST SSSI GESSSRRGEN TEST Vea seni = a 

1 2 3 4 5 6 | 7 8 9 0 | 11 12 | 18 14 | 15 

ot ee A ee Pee fe i pe a tf 
0 percent ____..__ = ee 2a ea a oa en a | ee | 
12.5 per cent ____- Solel Sei Piceite si lie ceil cre ome dt oka Swe eset lem Sefer I 
25 per cent -___-_- ap | seme sre te Seo ice tt se 0 0 0 0 0 0 0 0 0 
50 per cent ______- ar | Se | at | 0 0 | 0 0 | 0 0 | 0 0 | 0 0 | 0 0 


IX, B, 6 Ruediger: Germicidal Power of Glycerin ATT 


Tables XXIX to XXXVI show that when diluted with horse 
serum the germicidal power of glycerin is very feeble. In 12.5 
per cent of glycerin in horse serum, the typhoid bacillus, Staphy- 
lococcus albus, Staphylococcus aureus, the bacillus of anthrax, 
the bacillus of diphtheria, and the bacillus of glanders were alive 
at the end of fifteen days; the bacillus of plague was dead after 
ten days, and the spirillum of cholera was found to be dead on 
the fifteenth day. Twenty-five per cent glycerin killed the ty- 
phoid bacillus in fifteen days; Staphylococcus albus, Staphylococ- 
cus aureus, and the bacillus of anthrax were alive on the fifteenth 
day. The bacillus of plague lived seven days, the spirillum of 
cholera lived five days, the bacillus of diphtheria lived eleven 
days, and the bacillus of glanders gave negative results after 
six days. 

In 50 per cent glycerin the typhoid bacillus was alive at the 
end of twenty-four hours, but negative results were obtained 
thereafter. Staphylococcus albus lived five days; Staphylococ- 
cus aureus, ten days; and the anthrax bacillus was alive on the 
fifteenth day. At the end of one day, the bacillus of plague 
gave a positive result and the spirillum of cholera a negative 
result. The bacillus of diphtheria and the bacillus of glanders 
lived five days and three days, respectively. 


CONCLUSIONS 


Glycerin has a distinct, although feeble germicidal action. 

The germicidal action varies greatly with the temperature, 
being much feebler at a temperature of 15° C. than at from 
30° to 35° C. 

The germicidal action varies with the diluent employed; in 
glycerin diluted with physiologic salt solution the microdrganisms 
died much sooner than in glycerin diluted with bouillon or with 
horse serum. 

In dilutions up to 50 per cent, glycerin did not destroy the 
bacillus of anthrax in fifteen days. This may be due to the 
presence of spores. 

Glycerin seems to be a selective poison for the bacillus of 
plague, the spirillum of cholera, and the bacillus of diphtheria. 

In 50 per cent of glycerin in physiologic salt solution all the 
nonspore-forming organisms died in less than four days. 


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THE VITALITY OF THE CHOLERA VIBRIO IN MANILA WATERS 


By Otto ScHOBL 
(From the Biological Laboratory, Bureau of Science, Manila, P. I.) 


The character of the recent outbreak of cholera in Manila 
evidenced no indication of being one due to the spread of the 
infectious agents by means of the water supply. 

As water is one of the most important factors in the dis- 
semination of cholera, it was deemed of interest to arrange an 
orientation experiment as given below. 

In order to gain some information as to the vitality of cholera 
vibrios in Manila waters, the tap water and the sea water were 
considered. Sterile distilled water was included for comparison. 

It is evident from the literature,? as well as from theoretical 
grounds, that the vitality of cholera vibrios in water will 
be subject to variations owing to different factors, such as, 
the temperature of water, the chemical composition of water, the 
competition of cholera vibros with other bacteria present, 
the number of cholera vibrios, and the physical condition of 
the water. 

The consideration of the experimental evidence gives a certain 
general idea as to the vitality of cholera vibrios in water; but 
owing to the multiplicity of factors involved as well as to the 
differences in local conditions it is impossible to apply the re- 
sults of the experience gained elsewhere to local conditions in 
a particular place. Therefore, it was thought more instructive 
to arrange a simple experiment under such conditions as might 
occur in this community. 

The following experiments were made: 

1. Series of 5 test tubes containing 10 cubic centimeters of distilled water. 

2. Series of 5 test tubes containing 10 cubic centimeters of tap water. 

3. Series of 5 test tubes containing 10 cubic centimeters of sea water were 
inoculated in such a way that 1 loopful of cholera feces (typical rice- 


water stool) was planted in the first test tube, 3 loopfuls in the second, 
5 in the third, and so on. 


* The results of experiments by various authors regarding the vitality of 
cholera vibrio in water will be found tabulated in Kolle und Wassermann, 


Handbuch der pathogenen Microorganismen (1903), 1, 196. 
» 479 


A80 The Philippine Journal of Science 1914 


The tubes were thoroughly shaken, and a loopful was trans- 
planted from each tube into a test tube containing peptone solu- 
tion. Every tube gave positive growth of cholera vibrio upon 
transplant to a plate containing Dieudonné’s medium, showing 
that a sufficient number of cholera vibrios were inoculated into 
each tube of water to be found in1 loopful. The tubes containing 
water polluted with cholera feeces were allowed to stand at room 
temperature. Sunshine was excluded, but direct daylight had 
free access. 

From time to time a loopful of water was subplanted into 
peptone solution. From the time the cholera vibrio could no 
longer be recovered in a loopful, larger quantities were trans- 
planted. The details of the experiment are evident from Table 
I. Plates of Dieudonné’s medium were used exclusively. The 
experiment was begun in November, 1913, and the one hundred 
sixth day of the experiment was March 2, 1914; that is to say, 
the experiment was carried on during the latter part of the cool 
season. 
SUMMARY 


The table needs but a few remarks. It shows that the cholera 
vibrios disappeared from distilled water first and that they were 
evidently most numerous in the sea water. The room temper- 
ature (25° C.) was apparently very favorable. The unexpect- 
edly long period of time during which the cholera vibrios were 
found alive in the tap water shows that under conditions like 
those in tube I cholera vibrios may remain alive and multiply 
for a considerable length of time. It is also evident from the 
table that the vibrios remained alive in less-polluted water (tube 
I) longer than in the heavily polluted water (tube IX). 

The theoretical possibility in the Philippine Islands of intro- 
ducing Asiatic cholera from port to port by means of water 
carried on board of ships and of maintaining a source of infec- 
tion in waters polluted with human excreta thus finds experi- 
mental corroboration. 


Vitality of the Cholera Vibrio 481 


Schobl 


IX, B, 6 


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THE MORPHOLOGY OF THE ADULTS OF THE FILARIA FOUND 
IN THE PHILIPPINE ISLANDS 


By ERNEST LINWOOD WALKER 
(From the Biological Laboratory, Bureau of Science, Manila, P. I.) 


One plate 


Ashburn and Craig (1906) described the larval filarize found 
in the blood of a native of the Philippine Islands as a new species, 
Filaria philippinensis. The diagnosis of the new species was 
based on the study of the larve from a single case of infection. 
In 1907 these authors made a study of four additional cases of 
filariasis in the Philippines, which they decided were infected 
with the same species, and they performed some experiments on 
the development and transmission of this species by the mosquito, 
Culex fatigans. Ashburn and Craig distinguish Filaria philip- 
pinensis from the hitherto described species by certain peculiar- 
ities in the morphology and the lack of periodicity of the larve. 
Furthermore, they believe it to be the only indigenous species, 
because the previous descriptions of filarial larve observed in 
natives of these Islands contain nothing which would exclude 
the possibility that the observers were dealing with Filaria 
philippinensis. 

Phalen and Nichols (1908 and 1909) made somewhat extended 
observations on the occurrence and distribution of filariasis and 
elephantiasis in the Philippine Islands. They found both to be 
more prevalent and widespread than the observations of Ashburn 
and Craig would indicate. Moreover, they found a distinct noc- 
turnal periodicity in the appearance of the larval filariz in the 
peripheral circulation in all of their cases, and they concluded 
that the larve in all of the cases studied by them corresponded 
to the larve of Filaria bancrofti Cobbold. 

Recently Dr. J. W. Smith (1914), chief sanitary officer of 
Bilibid Prison, has had an extensive series of blood examinations 
made of the prisoners in that institution. These prisoners come 
from all parts of the Philippine Islands. The examinations were 
made of blood taken both during the day and at night. Filaria 
infection was found in about 6 per cent of the prisoners. The 
larval filarize have been found to exhibit a distinct nocturnal 


483 


484 The Philippine Journal of Science 1914 


periodicity in all cases. While elephantiasis or other clinical 
symptoms of infection with filaria are not common in this 
institution, they are by no means unknown. 

The identification of the species of the Philippine filaria both 
by Ashburn and Craig and by Phalen and Nichols was based 
upon the study of larval forms alone. Several pathologists work- 
ing in the Philippines, including Calvert (1902), Bowman 
(1911), and Crowell, have observed sections of adult filaria in 
the tissues, but so far as I am aware no intact adults have 
hitherto been discovered. ; 

I am indebted to Dr. J. W. Smith for the adult filarize de 
scribed in this paper which were obtained from a cyst removed 
surgically by his assistant, Doctor Manalak, from the inguinal 
region of a Filipino in the hospital of Bilibid Prison. This man 
was a native of Rizal Province and had, so far as known, never 
been outside of the Philippine Islands. Examinations of his 
blood for filarial embryos, both day and night, nine days before 
and on the day of the operation were negative. The cyst re- 
moved contained 1 adult male and 3 adult female filariz. One 
of the females was broken in removing it from the cyst; the 
other 2 females and the male were secured intact. 

The worms were washed in physiological salt solution, fixed 
in hot 70 per cent alcohol, and preserved in 70 per cent alcohol 
containing 5 per cent of glycerin. They were prepared for ex- 
amination by evaporating the alcohol from the glycerin-alcohol 
mixture and then mounting each in pure glycerin on a large slide, 
under a large cover glass supported by pieces of glass at each 
corner to prevent crushing the worm. 

The anatomy of these filariz, together with measurements, 
has been very thoroughly described, because it is believed that 
only by such careful descriptions can the identity of the filariz 
of different countries be determined. There is a noteworthy 
lack of careful and thorough work on the hitherto described 
adult filaria, and it is intended that the following description 
shall be accurate and complete for the species and serve for 
comparison with the other filarie. 


DESCRIPTION OF THE FILARIA: 
I. THE FEMALE 


General.—A long, slender, cylindrical, white worm. The an- 
terior end tapers slightly to a short neck, and terminates in a 


* Not yet published. 


1x,B,6 Walker: Morphology of the Adults of Filaria 485 


slightly bulbous head which is flattened anterioposteriorly 
(Plate I, fig. 1). The posterior end is straight or slightly curved. 
It tapers slightly to a bluntly rounded tail which is not bulbous 
(Plate I, fig.3). The lateral lines are not apparent. The cuticle 
shows exceedingly fine cross striations which vary in distinctness 
in different parts of the worm. The head bears laterally 2 
series of tiny papilla. No circumanal papille are apparent 
(Plate I, fig. 3). The measurements of the two intact females 
are as follows: 


TABLE 1.—H«xternal measurements of 2 female filarix. 


| Measurement. lis Now dt No. 2. 
mm mm | 
ACMI COTA renrns eee SESE eRe ILS eee eR eek Se see = Pe Cee eee ee ee 84.0 81.0 
“HD CO SG gee a een St Oe cae eae aaa oe ee Seas oe, Sees See ie 0.2268 | 0.2268 
TUDE eo NES oe OES eS Seen ee, See = ee sae ee ess 0. 063 0. 058 
LELTOEE UT LO) eles Se i ai ol oe ae he ih ONE oe Ren eS ee 0.0567 | 0.0518 | 
MimMetar OF Crd Gh Lakes) eee Reh EEE Se eee rk See 0.044 | 0.045 | 
Distance from head) to vulva == 2222 5 22 5s i ed eee is wt: _ Pe as a a i 0.729 0.79 
Wintance fromvanua to end of tails 2. =e ee 0.21 0.2 | 


Intestinal tract.—The mouth is terminal and minute, and does 
not appear to be surrounded by any appendages. There is a 
slight thickening of the cuticle lining the mouth (Plate I, fig. 2). 
It opens into a long slender csophagus, which is slightly en- 
larged posteriorly and which is from 1.13 to 1.34 millimeters in 
length. It is slightly constricted at its junction with the in- 
testine (Plate I, fig. 1). The slender intestine, which is of 
approximately uniform diameter, runs practically straight the 
whole length of the body to the anus, which opens on the ventral 
side at a point from 0.20 to 0.21 millimeter from the end of the 
tail (Plate I, fig. 3). 

Reproductive organs.—The vulva is situated anteriorly on the 
ventral surface, at a point from 0.729 to 0.77 millimeter from 
the anterior end. There is a slight protuberance at the opening 
which appears to be two lipped (labium). The vulva opens 
directly into the vagina, which is enlarged at its anterior end 
into a pear-shaped organ (receptaculum seminis) placed more 
or less obliquely and measuring 0.178 millimeter long by 0.06 
millimeter broad (Plate I, fig. 1). From the receptaculum semi- 
nis the vagina extends backward from 6.5 to 6.8 millimeters. 
At first straight and slender it makes one or two loops just 
behind the junction of the cesophagus with the mid-gut and 


" 486 The Philippine Journal of Science 1914 


gradually enlarges until it occupies almost the entire body cavity. 
In these specimens the enlarged syne ts part is filled with 
young larve. 

The posterior, enlarged, end of the wagina branches to form 
the two uteri (Plate I, fig. 4), which extend side by side straight 
backward nearly the whole length of the worm, occupying the 
entire body cavity except for the slender intestine; they are 
filled with larve and ova containing larve. At a distance of 
from 5.9 to 11.6 millimeters from the posterior end of the worm 
the uteri become abruptly contracted to be continued as the 
slender ovaries. The uteri do not always become constricted 
to form the ovaries at the same point. Thus in female No. 2, 
one uterus gives rise to the ovary at 8.4 millimeters and the 
other at 11.5 millimeters from the posterior end of the worm. 

The two ovaries form one or two loops upon themselves, and 
extend to within from 4.8 to 1.4 millimeters of the posterior end of 
the worm. The anterior part of the ovaries are empty tubes, 
but the posterior, distal, ends are filled with partially developed 
ova and undifferentiated protoplasm. 

The ovum is oval in outline, and contains the immature larva. 
The ova measure from 45 to 47 microns in length and from 27 to 
29 microns in breadth. 

The free intrauterine larve (immature) measure from 213 
to 233 microns in length, exclusive of the sheath, and from 4.7 
to 5.8 microns in breadth. The larva is inclosed in a sheath 
which extends beyond the caudal and cephalic ends and which is 
sufficiently loose to permit of forward and backward movements 
of the larva within it. The tail of the larva is tapering and 
pointed. 

Il, THE MALE 


General.—The male is shorter and slenderer than the female 
filaria. Ittapers slightly at both the anterior and posterior ends. 
The tapering anterior end is slightly enlarged into a bulbous 
head, which is flattened anteroposteriorly (Plate I, fig. 4) and 
bears laterally 2 rows of papille that are even more minute 
than in the female. The posterior end is spirally coiled two and 
one-half to three times (Plate I, fig. 5). There are no apparent 
lateral lines. The male shows transverse striations of the cu- 
ticle much less distinctly than the female; in fact, the striations 
are apparent only at certain curves of the coiled tail. There 
are 3 pairs of postanal and numerous pairs—at least 32—of 


Ix,B,6 Walker: Morphology of the Adults of Filaria 487 


preanal papilla, which are very small and difficult to distinguish 
(Plate I, fig. 5). 


TABLE II.—H ternal measurements of the male filaria. 


Millimeters. 
Length (exclusive of coiled tail) 33.0 
Greatest thickness 0.126 
Diameter of head 0.0486 
Diameter of neck 0.0437 
Diameter of end of tail 0.034 
Distance from cloaca to end of tail 0.129 


Intestinal tract.—The mouth, as in the female, is terminal. It 
opens into the long slender cesophagus of approximately uniform 
diameter, which extends straight backward 1.23 millimeters. 
_ It is slightly constricted at its junction with the mid-gut. The 
intestine is slender, of approximately uniform diameter, and runs 
nearly straight the whole length of the body of the worm. It 
enters the cloaca which opens on the ventral side of the worm 
at 0.129 millimeter from the posterior border of the curved tail. 

Reproductive organs.—The genital organs of the male consist 
of an elongated testis which is continuous with the spermatic 
duct and opens into the cloaca, the whole constituting a single 
cylindrical organ running ventrally nearly the entire length of the 
worm. The proximal end of the spermatic duct consists of a 
narrow, muscular ductus ejaculatorius, opening into the cloaca. 
The spermatic duct then becomes gradually dilated to form the 
seminal vesicle, which is from one-third to one-half of the di- 
ameter of the body cavity. This is continued into the slightly 
more dilated, long cylindrical testis which almost completely fills 
the body cavity and extends to within 1.7 millimeters of the an- 
terior end, or to within 0.469 millimeter from the posterior end 
of the esophagus. The distal (anterior) end of the testis tapers 
somewhat and terminates in a bluntly barbed end. The acces- 
sory genital organs consist of 2 spicular sacks situated laterally 
to the cloaca, containing spicules which are of unequal length 
(Plate I, fig. 5). 


COMPARISON OF THE PHILIPPINE WITH OTHER ADULT FILARIA 


The complete measurements of the adult Philippine filaria and, 
for comparison, the measurements, so far as they are given, of 
the hitherto described species of human filaria are given in 
Table ITI. 


1914 


SS. Se er ee RO ee ee ee ee ee oe a ee GEO Se Ek ee ae OCT OS" mee ion ewe ger ee [!8} Jo pus 03 snuy 
S snseydose jo yyZue7T 
38 [f8} JO pus Jo JoZONIvIGD 
9 ot PSAs ea ag a eee cae lie a meee ee | ime aioe “Gig ees sa ee age ooo ST0"Okge ie pee es ne ees yoeu Jorojowerq 
ee Eee eee TO'OR ot ae oy oe leo oa 630"Ome ea ily poor ee pay Jo raj9urBIG 
= S050) |00) -4210 5 Sennen cee 080 90°0 oro SS2E°0' <6) 0) 920 Ole eee ae ee een aes TozOUIVIP 48938915 
— CUS} a FA) tf: ie [ie = oe 008-0 “92 0°oF O°PP- 0°0F O°62510°00)  SONSRa ae eee er cons 2 Sa eee re qyizueT 
S PIB “SZ 
Ss ah = ES Se [f8} JO PWS 0} SolABAO Jo doo] Ao;10480g 
= GL L2a) Cale ess <a ee ae ae {eqn Jo yzZuUe'T 
eS 86'S SUON i Me ee ae ee BUISBA JO YIZUST 
Ny 00m Sk Ale aes eee SIUIWes UN[Noe}deoer Jo YIpEoAg 
& ee oe | eee RR Sami Sa (aie ings ante idee ea cae Neem ea S20 SS Ws Se SIUIWIES Wn[Ndededed Jo yABue'T 
= TL0 | 99°2 9L°0 2% 090 LO STcaPlnO) |e oe ae a A OS6CLO- es a eee os Lee peoy 03 BANA 
s CAs RSITSISO) < faR a ee ee a a ae SFT 0 8250) =BCGiO! | Se VG#0 SES iON, Te Oke (0850) oe eee aoe [83 JO pus 07 snuy 
RSet | SESS See a ee eT ae ae oem nae en me pce te Sal |e ac a snsBydose Jo yyZue'7 
[ae cere <3 eee SOKOcS See Geen Se ae CCUM Cs 7g a ges. a ye pes SFO "Oi PFOl0' |S a Gar ee [}8} JO pus Jo toJOWeIG 
SS) CO0KO wine ee ge ee el = ae #500 CRO'O ee) .  Geeaaer sere O90 Oss 90f0) | See ees aes see yoeu Jo za}0uBIG 
= SOLO! ea ae E'02=6010's<.)| iy as eee aes 10°0 QUONO Ss =v lee ee oe 890 OL2990%0 ie ea = es a a SOS ees P¥ey Jo JoJOUIeIG 
120 8°0 - 90 | 92°0 -12'0 Lg°0 ZrO 92 °0'=30850: |"920-0 =SE "On| Ze 0m os |e Shae a SS Joj}eUreIp 489} 8A15) 
(OST-98)0°I8 | 0°99T 0°98- 0°S9 0°0F-0 08 008-0 ‘OL 0°S6- 0°08 OPL9= 0 109) oe 'OFS= ONE B a oe a eo eee qj3ue'7 
‘o[BUlOT “T 
eRoonr | Monin" |“"chanar”?| “oS | “oct | “Ment! | mee na | su Sing | 
ie) 
Sor ‘unum Buyoafur aiojy ynpo fo siajzowyjuu m szuameimspayy— TIT WTav 


x,B,6 Walker: Morphology of the Adults of Filaria | 489 


While the measurements of filaria vary with the maturity of 
the specimens measured, and there exists considerable discrep- 
ancy in the dimensions of the same species as given by different 
authors, yet we can, I believe, at least exclude by these measure- 
ments the identity of the Philippine filaria with Filaria (Loa) 
loa and Filaria magalhaesi. Filaria (Loa) loa is further ex- 
cluded by the presence of rounded bosses on the cuticle, by the 
absence of a spirally twisted tail in the male, by the presence 
of 4 well-marked papillze of peculiar form on each side of the 
ventral surface of the tail, and by the relatively great distance 
of the vaginal opening from the anterior end; and Filaria magal- 
haesi, by the habitat of the adult in the left ventrical of the 
heart, by the presence of 4 pairs of preanal and 4 pairs of post- 
anal papillze in the male, and by the relatively great distance 
of the vaginal opening from the anterior end. 

Filaria perstans is excluded by the triangular cuticular append- 
age of the tail, by the absence of 2 unequal spicules in the male, 
and by the characters of the larvee, which are small and blunt- 
tailed and possess no sheath. 

Filaria demarquayi differs from the Philippine species in the 
greater diameter of the head, in the tail which tapers abruptly 
behind the anal papille, in the marked cuticular thickening at 
the tip of the tail, and in the small size and absence of a sheath 
in the larva. 

Filaria ozzardi is excluded from the identity of our spécies 
by the bulbous end of the tail and by the small size and absence 
of a sheath in the larva. 

The Fijian filaria is haliteed by Bahr (1912), who has recently 
investigated filariasis in the Fiji Islands on a commission from 
the London School of Tropical Medicine, and by Leiper, helmin- 
thologist of the London School of Tropical Medicine, to whom 
adult worms were submitted, to be Filaria bancrofti, in spite 
of the absence of periodicity in the peripheral blood displayed 
by the larve. Bahr believes that this lack of periodicity of the 
filarial larve in the Fiji Islands is due to a partial adaptation 
of the parasite to its commonest transmitting host, Stegomyia 
pseudoscutellaris, a mosquito which feeds by day only. 
the Philippine filaria is identical with Filaria bancrofti or (2) 

Therefore, there remain two possibilities; namely, (1) that 
that it is a new species, as was believed by Ashburn and Craig. 
I believe that the former possibility is the truth. 

There are no characters that preclude the identity of the Phil- 
ippine filaria with Filaria bancrofti. The small size of the 
intrauterine larve is evidently due to their immaturity. On the 


490 The Philippine Journal of Science 


other hand, all of the essential characters of the Philippine filaria 
correspond to those of Filaria bancrofit. The more important 
of these characters are: (1) General, (a) size, (b) shape, (c) 
head bearing 2 series of tiny papille, (d) slight thickening of 
the cuticle lining the mouth, (e) delicate cross striations of the 
cuticle which vary much in distinctness; (2) female, (a) posi- 
tion of the vulva, (6) the pyriform enlargement of the terminal 
portion of the vagina (receptaculum seminis), (c) the absence 
of anal papille; (3) male, (a) the tendrillike coiling of the 
tail with an incurved end, (b) 2 dissimilar spicules, (c) the 
presence of 3 pairs of postanal papillz and their size, shape, 
and position, (d) the presence of numerous (about 32) pairs 
of minute preanal papille; and (4) larva, (a) a pointed tail 
and (b) the possession of a sheath. In the literature available 
here, I have been unable to find any detailed description of the 
internal anatomy of Filaria bancrofti; therefore, I am unable to 
compare the details of the anatomy of this species with that of 
the Philippine filaria. However, on the basis of external char- 
acters, especially of head and tail and their appendages, which 
are generally used for classifying adult filaria, the Philippine 
filaria appears to be identical with Filaria bancrofti. This deci- 
sion, based on the study of these specimens from one patient, 
does not preclude the possibility of another species of filaria 
existing here, but in conjunction with the evidence of recent 
studies of the larval forms it renders it extremely improbable, 
and it establishes definitely for the first time the existence of 
Filaria bancrofti in the Philippine Islands. 


LITERATURE REFERRED TO IN THE TEXT 


ASHBURN, P. M., and Craic, C. F. A new blood filaria of man: Filaria 
philippinensis. Am. Journ. Med. Sci. (1906), n. s. 132, 485-448. 
IpEM. Observations upon Filaria philippinensis and its development in the 

mosquito. Phil. Journ. Sci., Sec. B (1907), 2, 1-14. 

Baur, P. H. Filariasis and elephantiasis in Fiji. Journ. Lond. School 
Trop. Med. (1912), Suppl. 1. 

Bowman, F. B. A case of dysentery caused by Balantidium coli with 
coincident filarial infarction of the spleen. Phil. Journ. Sci., Sec.. B 
(1911), 6, 147-158. 

CALVERT, W. J. A preliminary report on the blood in two case of filariasis. 
Bull. Johns Hopkins Hosp. (1902), 13, 23. 

LEIPER, P. T. See Bahr, 1912. 

PHALEN, J. M., and NicHous, H. J. Filariasis and elephantiasis in southern 
Luzon. Phil. Journ. Sci., Sec. B (1908), 3, 293-3038. 

IpEM. The distribution of filaria in the Philippine Islands. Jbid. (1909), 
4, 127-135. 

SMITH, J. W. Some clinical observations on filariasis in Bilibid Prison. 
Read before the Manila Medical Society, April 6, 1914. 


ILLUSTRATIONS 


PLATE I 


(The figures in Plate I, with the exception of fig. 2, were all drawn with a Zeiss AA objective, 
$ ocular, and tube length of 160 millimeters, and with the aid of a camera lucida. Fig. 2 
was made with a Zeiss DD objective. The figures are all reduced to one-fourth size of 
drawings. The drawings are somewhat diagrammatic in order to bring out clearly the 
essential structures. Drawn by T. S. Espinosa.) 

Fig. 1. Cephalic end of female of Filaria bancrofti, showing the anterior 

portions of the alimentary tract and generative organs. 

2. Cephalic end of the female of Filaria bancrofti, more highly mag- 
nified, showing the details of the structure of the head. 

8. Caudal end of the female of Filaria bancrofti. Note the position 
of the anus and the absence of anal papille. . 

4, Cephalic end of the male of Filaria bancrofti. Note the constriction 
of the alimentary tract at the junction of the csophagus with 
the mid-gut. 

5. Caudal end of the male of Filaria bancrofti. Note the tendril- 
like coiling of the tail, the three postanal and the numerous 
minute preanal papille, the spermatic duct (dorsal) and the 
intestine (ventral) opening into the slightly dilated cloaca, the 
opening of the cloaca, and the extended spicule (the other shorter 
spicule lies under the one shown in the figure). 

491 


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Se acl aed 


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foots ws Wha Pe Yoyatin Wu e: la af 
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~ 


WALKER: MORPHOLOGY OF FILARIA. | [Pu. Journ. Sct., IX, B, No. 5. 


Fgi. 1. Cephalic end Fig. 3. Caudal end of Fig. 4. Cephalic end of 
of female. female male. 


Fig. 2. Cecephalic end of Fig. 5. Caudal end of 
female. male. 


PLATE |. MORPHOLOGY OF FILARIA BANCROFTI. 


i 
4 
. 
“ 
. 
ha 
4 f 2 
x 
‘ 
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me 


PELVIMETRY AND CEPHALOMETRY AMONG FILIPINAS 


By Honoria AcosTA-SISON 


(From the Department of Obstetrics, College of Medicine and Surgery, 
University of the Philippines) 


Practically no previous systematic measurements of the pelvis 
in Filipinas have been performed, those cases only having been 
measured in which the physician was confronted with a gross 
pelvic abnormality from osteomalacia or rickets or with a case 
of difficult labor. Apparently, also, no studies have been made 
of the relationship in size between the pelvis of the Filipino 
mother and the head of the infant. This may be explained by 
the facts that until recently untutored midwives had almost the 
entire control of the maternity cases; that the prospective 
mothers traditionally do not consult a physician until the mid- 
wife has failed to bring forth the baby and the patient is near 
death; and that until two and a half years ago there did not 
exist large charity hospitals with modern equipment and facil- 
ities such as are now found in the Philippine General Hospital. 

It has been asserted by different investigators, such as 
Stratz,(9) Topinard,(10) Waldeyer,(11) Riggs,(6) and others, 
that the pelvis of the less civilized races is narrower and deeper 
than that of the Caucasian race; and writers, such as Engel- 
man,(2) Scharlau,(8) Ploss,(5) and Gache,(3) have observed the 
comparatively easy labors of primitive people. 

The literature concerning the Filipino pelvis consists of a 
description of a case of osteomalacia by Baldomero Roxas, (7) 
and one of an osteomalacic pelvis by José Montes.(4) However, 
there are no articles dealing with the normal Filipino pelvis. 

Purpose.—The present study has been undertaken with the 
purpose of investigating the Filipino pelvis and its relationship 
to the child’s head and also to establish a comparison between 
the Filipino and European pelvis. It was attempted at the sug- 
gestion and with the assistance and advice of Dr. Fernando Cal- 
deron, who kindly consented to measure the diagonal conjugate 
of all the cases reported below. 

493 


494 The Philippine Journal of Science 1914 


Material.—The material on which this study is based consists 
of the records of 181 women and 117 babies who were patients 
in the obstetrical department of the Philippine General Hospital 
during the period from November, 1913, to February, 1914. To 
insure accuracy in figures, only the cases personally measured 
by us have been used. There have been eliminated 2 cases of 
generally contracted pelvis, 2 cases of flat pelvis, and all cases 
in which labor resulted in premature or multiple babies. The 
figures of Riggs, who has studied this subject extensively, have 
been used to establish a comparison between the European or 
American pelvis and that of the Filipina. Riggs’s normal meas- 
urements were averaged from 707 white women and children. 
The diameters of the pelvic outlet, however, have been taken 
from Edgar,(1) as they have not been included in Riggs’s work. 

Method of measurement.—The measurements of the external 
diameters have been taken with the tips of the pelvimeter placed 
just opposite the bone and not in the outer or inner lip of the 
ilium. In taking Baudelocque’s diameter, the closest proximity 
to the bone was attempted and the anterior tip of the pelvimeter 
was placed on the upper border of the symphysis pubis rather 
than on its anterior surface. The anteroposterior diameter of 
the pelvic outlet has been measured from the lower border of 
the symphysis pubis to the sacrococcygeal articulation. These 
points were chosen because the coccyx is movable, at least in 
our reported cases, and they, therefore, practically represent the 
shortest distance in the anteroposterior diameter of the pelvic 
outlet. 


TABLE I.—Comparison of the pelvic measurements in American and 
Filipino women. 


pp es Le ees ON Pade Se 


Ameri- 


Measurement: ean, 707 | Filipino, | Differ- 

cases |181 cases.| ence. 
(Riggs). | 

cm. cm. cm 

SING 2c ed eh i eee ode er nt ee ee eee 25.47 23.9 1,47 
Cresta 27.998 24.91 3.088 | 

Trochanters, 625536 5-3 Wes Jans. eee a. res le. Sy 30. 90 28. 103 2.797 

Baudelocque’s 19. 71 17. 63 2.08 

Diagonal conjupate 222 ssc) cere ae cn ee eee 12.26 12.00 0.26 

' Anteroposterior diameter of the pelvic outlet______________________ 912.5 10. 05 2.44 
| Transverse diameter of the pelvic outlet ________-__________________ 11.00 10,00) be Sete 


& Edgar. 


x, B,6 Acosta-Sison: Pelvimetry and Cephalometry 495 


TABLE I].—Comparison of the cephalic measurements in American and 
Filipino infants. 


Ameri- | 
Measurement. can, 707 | Filipino, | Differ- 
eases (117 cases.| ence. 
(Riggs). 
cm. cm. cm 
Occinitomentale eee re ee ee 18.33 12.11 1292). | 
Grecinitonrontal es ne eee oe eo ae oh ee gees Sema ses 11. 707 10. 96 0. 747 | 
Suboccipiaprem maior oe ares eh hs Bed ee 9.70 9.28 0.42 
VOPR Gar Be oes Soe Cee eee a ee ee et oe Pa a SI 9.25 8.63 0.62 
ROM ONARRe ener oo peace et ees 8.00 6. 829 1.171 
CICCMNIOIENTCIFCUMIerence!.- 2-5 -=-- 22 se A ee Age (RE ee 
Oaeipitofrontal circumference ---.-.=----.----1+--=----<-----s-=-+2]---.-2---- S204? fe= 222 -20Re 
Suboccipitobregmatic circumference_-__-.__.-_._-.----_---------_-- 32.17 30.26 1.91 


Table I shows that the diameters of the Filipino pelvis are 
shorter than those of the American or European pelvis. On 
studying this table closely, one finds that the relation of one 
diameter to another is altered in such a way that the propor- 
tion of the crests with the spines in the Filipino pelvis is much 
smaller than the proportion of the same diameters in the Euro- 
pean or American pelvis and that the proportion of the diagonal 
conjugate with the other diameters in the Filipino is very slightly 
larger than in the American. This would seem to show that the 
Filipino pelvis is narrower and relatively slightly deeper than 
the American pelvis. This diminution may be explained by the 
small stature of the Filipinas. However, the most striking dif- 
ference is that, although the Filipino pelvis is smaller than the 
American pelvis, the transverse diameter of the pelvic outlet 
is the same as that of the Caucasian race. 

To find an explanation for this peculiarity would take us back 
to the mechanical theory of the production of the shape of the 
human pelvis. It is well known that the pelvis of the child alters 
in shape in proportion to mechanical] influences such as that of 
osteomalacia. It is also an established fact that the sitting 
posture flares out the ischial tuberosities, and this is proved in 
some cases of osteomalacia where the patient has maintained 
the sitting posture for a long time. In applying this theory in 
our cases, it must be remembered that the Filipino woman from 
her childhood has habitually accustomed herself to the squatting 
position or to sitting on the hard floor with the knees drawn up, 
and her occupation is such that she is obliged to be in this position 
for nearly the whole day. Whether she sews, cooks, washes, 
or sells in the market, she nearly always assumes this position. 


496 The Philippine Journal of Science 1914 


The squatting position has a greater tendency to flare out the 
ischial tuberosities than has the ordinary sitting posture on 
the chair, for in the former not only the body weight but also 
the weight of the thighs is transmitted to the ischial tuberos- 
ities. Perhaps the custom of carrying the child astride the 
mother’s or nurse’s hip may also have an effect toward the en- 
largement of the transverse diameter of the pelvic outlet. 

Table II shows the cephalic diameters of the Filipino and 
American child. As may be noticed, all cephalic diameters of 
the Filipino child are smaller by slightly over 0.5 centimeter 
to compensate for the undersized pelvis. Moreover, the oc- 
cipitomental and bitemporal diameters are over 1 centimeter 
shorter. The diminution of the cephalic diameters may also 
be explained, aside from the pelvic accommodation, by the small 
stature of the Filipinas. It has been asserted by most observers 
that the child of the primipara is smaller than are those of the 
multipara, but in our work we have made no attempt to deter- 
mine this point. 

CONCLUSIONS 


From the above findings, it seems justifiable to establish the 
following conclusions: 

1. That the Filipino pelvis is smaller than the American or 
European pelvis and that this may be explained by the smaller 
stature of the Filipinos as a race. 

2. That the transverse diameters of the Filipino pelvis, except 
the transverse diameter of the pelvic outlet, are smaller than 
those of the European or American pelvis. 

3. That the relative enlargement of the transverse diameter 
of the pelvic outlet may be explained by the habitual squatting 
-_position that the Filipino woman assumes and by the fact that 
the carrying of the child astride the mother’s or nurse’s hip may 
also affect the child in the transverse diameter of its pelvic outlet. 

4, That the head of the newborn is proportionate with the 
Filipino pelvis. 

5. That the cephalic diameters of the Filipino child are smaller 
than those of the American child by a little over 0.5 centimeter, 
with the exception of the occipitomental and bitemporal di- 
ameters, where the shortage is over 1 centimeter. 


REFERENCES 


(1) Epear, J. C. The practice of obstetrics. P. Blakiston’s Son & Co., 
Philadelphia (1907). 

(2) ENGELMAN, G. J. Labor among primitive peoples (1883). Cited by 
Riggs. 


x,B,6 Acosta-Sison: Pelvimetry and Cephalomeiry 497 


(8) GacHE, S. La rachitism en Amérique et son influence obstetricale. 
Ann. gyn. et dobst. (1903), 60, 175. Cited by Riggs. 

(4) MontsEs, Joss. Rev. Filipina med. y farm. (1912), 3. 

(5) Pioss, H. Das Weib in der Natur und Volkerkunde. IV. Aufl. 

(1895), 1, 184. Cited by Riggs. 

(6) Rices. A comparative study of white and negro pelves with a con- 
sideration of the size of the child and its relation to presentation 
and character of labor in the two races. Johns Hopkins Hospital 
Rep. (1904), 12, 421. 

(7) Roxas, BALDOMERO. Un caso interesante de seccién cesarea clasica 
por pelvis osteomalacica. Rev. Filipina med. y farm. (1911), 2, 1. 

(8) ScHaRLAv, B. Das Australier-Becken, Berlin (1903). Cited by Riggs. 

(9) Stratz, C. H. Die Frauen auf Java, Stuttgart (1907). Cited by 
Riggs. 

(10) TopinaRp. Des proportions générales du bassin chez l’homme, etc. 

Bull. Soc. danthrop. (1875), 504. Cited by Riggs. 

(11) WALDEYER. Cited by Riggs. 

129562——3 


be mi bees’, 
ro ee 
* 
ea 


so “tpi ae 


A NOTE ON THE PHYSIOLOGICAL ACTION OF THE PROTEOSES 


By R. B. GIBSON 


(From the Department of Physiology,’College of Medicine and Surgery, 
University of the Philippines) 


In mixed proteoses obtained by the gastric digestion of pure 
proteins, I observed (in 1902) that the filtrate from the hot 
alcohol-precipitated preparation still contained much proteose. 
The effect of such alcohol-soluble proteoses (ovalbumoses, ede- 
stinoses, caseoses) on the heart and on body temperature have 
already been reported. 

While preparing, on another occasion, the peptic proteoses of 
erystallized ovalbumin, the hot alcoholic filtrate from the dialized 
and concentrated proteoses was left on the laboratory table. 
On cooling, the precipitate had a peculiar granularlike appear- 
ance. Under the microscope, it was observed that the precip- 
itate consisted of minute spherules which resembled closely those 
found in the intermediate phase of the slow crystallization of. 
ovalbumin. A barely sufficient amount of this material was 
obtained to try the effect on blood pressure with a very small 
dog. The fall of blood pressure typical of proteoses was noted, 
but blood clotting was not effected in this experiment. 

This new method for the preparation of certain proteoses 
has made it of interest to study the physiological action of the 
products so obtained. Particularly is this so, as Pick and Spiro? 
and Popielski‘ and his associates have indicated that the so- 
called “peptone” shock of the earlier writers may be due to some 
impurity in the proteoses employed. Underhill® has shown that 
Pick and Spiro’s conclusions are untenable. However, their 
work is still quoted in much of the literature as final,* and seems 


* Gibson and Schultz, Journ. Pharm. & Exp. Therap. (1909-10), 1; Gibson, 
This Journal, Sec. B (1918), 8, 475. 

* Hofmeister, Zeitschr. f. physiol. Chem. (Hoppe-Seyler) (1889), 14, 165. 

* Ibid. (1900), 31, 235. 

“Arch. f. d. ges. Physiol. (Pfliiger) (1907), 120, 451; zbid. (1908), 121, 
239; ibid. (1909), 126, 483; also Czubalski, ibid. (1908), 121, 395; Gizelt, 
ibid (1908), 123, 540; Popielski, Zeitschr. f. Immunitdtsforsch., Orig. 
(1918), 18, 562. See also Zuntz, E., Compt. rend. Soc. biol. (1913), 73, 50. 

°Am. Journ. Physiol. (1903), 9, 345. 

* Cf. Cecil, ibid. (1911), 24, 156. 

499 


500 The Philippine Journal of Science 1914 


to be substantiated by Popielski who ascribes the active physio- 
logical principle of Witte’s peptone to an alcohol-soluble im- 
purity, “‘vasodilatine.” 

A study of the alcohol-soluble proteoses should go far toward 
clearing up the problem as to whether “peptone” shock is really 
due to the proteose itself or to alcohol-soluble impurities. It 
would be expected a priori that, if such an alcohol-soluble sub- 
stance is really present with the proteose, the process of separa- 
tion with alcohol would yield a nonactive product or that the 
active principle, if it is deposited on cooling, would be much more 
concentrated. If the physiological effects are essentially un- 
changed, it would be justifiable to conclude that these are charac- 
teristic of the proteose per se. 

Accordingly, 500 grams of Witte’s peptone were extracted with 
about 3 liters of hot 80 to 85 per cent alcohol. The alcohol was 
filtered off on a hot water funnel, allowed to cool slowly, 
and then placed in the ice box. A small amount of proteose 
separated out with the characteristic spherule formation. This 
residue was filtered, pressed out between dry filters, and pre- 
served. The filtrate was poured over the original proteose 
residue, and the procedure was repeated. After numerous ex- 
tractions, the residues were redissolved in water and dried on the 
water bath; about 60 grams of hot alcohol-soluble proteoses 
(preparation 1) were obtained. 

The reéxtraction of the original residue with fresh alcohol was 
then undertaken, and about 50 grams more of the alcohol-soluble 
proteoses were obtained. This material was again redeposited 
from a repeated hot alcoholic extraction until about 40 grams 
of proteose (preparation 2) were recovered. After every ex- 
traction, the deposit was spherular in character. 

The yield from Witte’s peptone was then over 20 per cent. 
The final products were obtained as fine white powders, sup- 
posedly proteoses. They give a typical (not pink) biuret reac- 
tion, a Millon’s test, a strong Hopkins-Cole reaction, and contain 
a small amount of loosely combined sulphur. They are com- 
pletely soluble to almost colorless solutions in hot water, but 
concentrated solutions yield a slight deposit of spherules on cool- 
ing and standing. They are precipitated by absolute alcohol. 
Unlike other proteoses or proteins, they may be dried on the 
water bath to an easily friable residue. The process of separa- 
tion is a tedious one; nearly three weeks were required to com- 
plete the above preparations. 

When injected intravenously into dogs, both preparations 


Ix, 8,6 Gibson: Physiological Action of the Proteoses 501 


produced the typical fall in blood pressure and inhibition of blood 
clotting. The amounts necessary to give these results are the 
same as those usually described (0.3 to 0.5 gram per kilogram 
of body weight).? No essential difference was noted between 
the effects of preparation 1 and the purer preparation 2. 

Proteose preparation 1.—June 24,1914. Dog, male, weighing 
4.9 kilograms. Blood samples obtained at 10.30 a. m. clotted in 
fifteen minutes. Blood pressure at 10.39 a. m. was 130 milli- 
meters. At 10.40 a. m., the dog received an injection into the 
saphenous vein of 2 grams (in 20 cubic centimeters of water), 
or 0.4 gram per kilogram of body weight of the alcohol-soluble 
proteoses. Blood pressure fell rapidly at first and then slowly 
to 30 millimeters of mercury; at 10.45 a. m. it was 50 millimeters 
only. Blood samples collected in duplicate at 10.45 a. m. and 
10.50. a. m. failed to clot. Blood pressure at 11.00 a. m. and 
11.05 a. m. had risen to 62 and 92 millimeters of mercury. 
Blood samples obtained at 11.26 a. m. clotted after twenty-five 
minutes. 

Proteose preparation 2.—June 25, 1914. Dog, female, weigh- 
ing 8.2 kilograms. Blood samples obtained at 12.03 p. m. clotted 
in fourteen minutes. Blood pressure at 12.04 p. m. was 148 
millimeters of mercury. At 12.05 p. m. the dog received an 
intravenous injection as above of 3.5 grams of the proteoses 
(0.4 gram per kilogram of body weight) in 40 cubic centimeters 
of water. Blood pressure fell regularly to 24 millimeters of 
mercury. At 12.06 p. m. the blood pressure was still 24 milli- 
meters of mercury. Blood samples obtained at 12.10 p. m. and 
12.15 p. m. failed to clot. Blood pressures at 12.20 p. m., 12.25 
p. m., and 12.30 p. m. were 60, 72, and 100 millimeters of mer- 
cury, respectively. Blood samples collected at 12.36 p. m. still 
failed to clot. 

CONCLUSIONS 


1. The preparation of semicrystalline alcohol-soluble proteoses 
from Witte’s peptone is described. 

2. When injected intravenously into dogs, these proteoses 
produce the typical fall in blood pressure and inhibition of blood 
clotting which have been repeatedly described for other proteose 
preparations. 


* Schmidt-Miilheim, Arch. f. Physiol. (1880), 33; Fano, ibid. (1881), 277; 
Chittenden, Mendel, and Henderson, Am. Journ. Physiol. (1899), 2, 149; 
Underhill, loc. cit., and others. 


502 The Philippine Journal of Science 


3. The amount of the alcohol-soluble proteose necessary to 
produce these effects (0.4 gram per kilogram of body weight) 
shows that in the process of separation there has been neither 
concentration nor loss of the active physiological principle. The 
more purified proteose was not less effective than the cruder 
preparation. This action must then be ascribed to the proteoses 
themselves and not necessarily to an alcohol-soluble impurity. 

I with to thank Dr. Isabelo Concepcién for the assistance which 
he has rendered me both in the preparation of the proteoses and 
in the carrying out of the blood-pressure experiments. 


THE LYMPHAGOGIC ACTION OF THE PHILIPPINE MANGO, 
MANGIFERA INDICA LINN4ZUS 


By R. B. Gipson and ISABELO CONCEPCION 
(From the Department of Physiology, College of Medicine and Surgery, 
University of the Philippines) 

The transient rashes occurring in the Philippines during the 
hot season are often popularly ascribed to eating the Philippine 
mango, Mangifera indica L., which ripens at this time. The 
mango seems to intensify the ordinary “prickly heat” symptoms, 
and individuals in the Islands who have a tendency to this affec- 
tion often find relief when they eliminate the fruit from their 
dietary. We have also had occasion to observe that the nursing 
child may be affected when mangos are eaten by the mother; 
these observations will be reported in a separate paper by one 
of us (Concepcién). 

The almost universal distribution of the mango in tropical 
countries and the large part that the fruit shares in tropical 
dietaries make the problem of “mango rash” an important one. 
Such rashes are commonly ascribed to the mango by the Philip- 
pine practioners of medicine, but we have been unable to find any 
description of these effects in the literature. The present inves- 
tigation was carried on to get some experimental evidence as to 
whether or not the mango is to be classed with such rash-pro- 
ducing substances as crustaceans, mollusks, etc., which Heiden- 
hain: designated physiologically as lymphagogues of the first 
class. 

The lymphagogues of the first class include “peptone,” albu- 
men, extracts of liver and intestine, and especially extracts of 
crustaceans, mollusks, and leeches. The physiological effects are 
a marked fall in blood pressure, an increased flow of lymph 
richer in solids than the normal, an inhibition of the clotting 
power of the blood, deep narcosis, anuria, and increased secretory 
action of the pancreas, salivary glands, and liver. Subsequent 
injections are progressively less effective. The apparent similar- 
ity of the effects of the lymphagogues of the first class with 
anaphylactic shock is of extreme interest. This similarity has 
been repeatedly pointed out. 


*Arch. f. d. ges. Physiol. (Pfliiger) (1891), 49, 209. 
503 


504 The Philippine Journal of Science 1914 


Rashes sometimes occur after the ingestion of strawberries, 
and a lymphagogic effect for these has been demonstrated by 
Clopatt ? and by Mendel and Hooker.? Our work has shown con- 
clusively that the mango is also to be included with Heidenhain’s 
lymphagogues of the first class. 

Dogs, anesthetized with ether only, were used in the 14 ex- 
periments performed. Extracts of the dried mango pulp (3 
experiments) were less effective than the strained and centrifu- 
galized raw juice which we employed in the remainder of the 
series. Mendel and Hooker arrived at a similar conclusion with 
strawberry extracts. Lymph was collected from the thoracic 
duct. The dogs had not been fed since the day preceding. In- 
jections of the mango juice, at 30° C., were made from a burette 
into the saphenous vein. Determinations of the total solids of 
the lymph, collected over ten-minute periods, were made in 6 
experiments. The samples were dried on the water bath and 
then heated in the oven at 105° C. until the weight was constant. 
Blood pressures were recorded graphically with a mercury mano- 
meter in the usual way. Blood samples (about 5 cubic centi- 
meters) were collected in test tubes from the femoral artery, a 
clean, dry, glass cannula being employed each time. Sufficient 
blood to wash out the cannula was allowed to pass through before 
the sample (in duplicate) was taken. The blood was considered 
to have clotted when the tubes could be reversed without spilling. 

The results of two of the experiments in which the total solids 
of the lymph were determined are given in Tables I and II. 
These experiments show an increased flow of lymph of almost 
three times the normal. This lymph is richer in solids than the 
samples collected before the injection. Blood pressure under- 
went the typical fall to be expected from lymphagogic substances. 
The clotting time of the blood is slightly extended in experiment 
10 (Table I), but is shortened in experiment 11 (Table II). We 
have observed this unexpected result in several of our experi- 
ments. Every sample of lymph which we collected promptly 
clotted.‘ 

That the mango juice may produce the typical inhibition of 


2 Skandin. Arch. f. Physiol. (1900), 10, 403. 

*Am. Journ. Physiol. (1902), 7, 380. 

‘Experiments have been reported in dogs with thoracic fistula in which 
the clotting time of the blood was only slightly, if at all, affected by proteose 
injections, while the coagulability of the lymph was delayed. Spiro and 
Ellinger, Zeitschr. f. physiol. Chem. (1897), 23, 185; Chittenden, Mendel, 
and Henderson, Am. Journ. Physiol. (1889), 2, 142. 


e 


Ix, B, 6 Gibson & Concepcion: Lymphagogic Action of Mango 505 


blood clotting is shown in the following experiment in which no 
thoracic fistula was made: 

Experiment 11, June 25, 1914.—Male dog, weighing 5.8 kilo- 
grams. Normal blood obtained at 9.40 a. m. clots in fifteen 
minutes. At 9.45 a. m. the dog received a rapid injection of 40 
cubic centimeters of fresh mango juice. Blood pressure fell 
immediately from 178 to 45 millimeters. Blood samples ob- 
tained five, ten, and thirty minutes after injection failed to clot 
in twenty-four hours. 

That a certain degree of tolerance or immunity results from 
consecutive injections, both for lymph flow and blood pressure, 
is shown in experiment 12 (Table III). In this experiment only 
from 17 to 20 cubic centimeters of the juice were given at a 
single injection, as the dogs do not withstand very well the 
repeated administration of larger amounts. In the latter case, 
the second injection may be nearly as efficient as the first, as 
shown in experiment 8 (Table IV). 

Clopatt has shown that the quantities of sugar and salts, 
in the berry extracts employed, were too small to ascribe the 
marked results obtained to a lymphagogic effect of the second 
class (Heidenhain). Mendel and Hooker calculated that the 
maximum amount of sugar used in the largest injection of straw- 
berry extract would not exceed 0.2 gram per kilogram of 
body weight of the dog used. The mango pulp has the following 
composition : 


Composition of mango pulp.* 
Per cent. 


Water 82.8 
Solids 17.2 
Sugar (as invert sugar) 13.24 
Acid (as citric acid) 0.18 
Protein 0.22 
Crude fiber 2.6 
Ash 0.45 


The sugar given in experiment 10 (Table I) amounts then 
to only about 0.3 gram per kilogram of body weight. Further- 
more, the increase in total solids of the lymph collected after 
' the injection of the mango juice is characteristic of the first 
and not of the second class, or crystalline, lymphagogues. With 
the second class, in fact, there is usually a diminution of the 
total solids. The additional evidence of the constant fall in 
blood pressure, the observation that the mango juice may produce 


* Pratt and del Rosario, This Journal, Sec. A (1913), 8, 59. 


506 The Philippine Journal of Science 1914 


the characteristic inhibition of the clotting powers of the blood, 
and the diminished response to consecutive injections indicate 
that the effects of the mango are similar to those of lympha- 
gogues of the first class. 


TABLE I.—Experiment 10, June 14, 1914. Male dog, weighing 11.9 


kilograms. 
Pree das 
Lymph 
Time. i A nd ee oe et Blood Remarks. 
baal ad | 
hw iee: Per cent.! mm. Min. 

2017-10-27 ae om Se 4.5 GiBite See tn es ee The lymph clots. 

NOS A PIN oe ce a pa ee a eed es cen eee eee 11 
10:b1-asan) 22 A eR ee ee 1 0) | (eae os Bae Injection of 25 cc. of fresh 
| | mango juice. 

1O:b2: asm ae sec ~ ee Soe eee |e aa ae (J) ee ee 
| 10.52-11.02'a. m_____. 12.5 US SUN yee Sa The lymph clots. 
| CLOG Aree carn ese | Pe | eee 8 \p arent 11.5 
| PATOLGS evar) Sea cto toate Bese e oa fice cee 12.0 

UL Ob amen eee foe eof SS lage reece 15.0 
}) 112-14. 22'a5 mes =| 5.0 | LP Se) Seg a ME eye Do 
| TOR at rn ee! sak ee ease one rhitey lisse ts be 

12.01-12.11 p. m____--- 4.5 Beth ere ate als oe ae rec uy 
| 12.12) pitino Sr eS oe eae coe 116 | eee eee | 

r= J 


TABLE I].—EHxperiment 14, July 3, 1914. Female dog, weighing 6.6 


kilograms. 
yaa alate Tas aap ed 
ymp 

| Time. _{. 20 9 grits favenetvel |Meat: Remarks. 
| x 1 Bil | Et Pk a 
/ cc Per cent.' mm Min 
| 10.51-11.01 a. m..-...- | 5.5 pai) | Sse Ss es i i cet el The lymph clots. 
| 1050 /a.pmn ae ese oer Re ae, eee Men meee Mae aa 14 
. TED eh, Mo eet ee eee eee, eae TAO) More ee wen Injection of 40 cc. of fresh 
| | mango juice. 
| 11.16-11.26a,m_......| 16.0 BTA a. yack Sask ees The lymph clots. 
Ss GA aa eee Bee Nie fe anh 66 |e -ee eee 

NE TRS se ee Pg SE he ete eel ee 6 
| 11,27-11.87 a. m_______ 18.0 BSGh| oo tens ee | ea ee Do. 
Hdceoial anit. °. Sse eel oe pW Ses) 134 9 | 
(GO a. inten = Pe ee Se ee ele 7 
| 11.40-11.50 a. m______- 11.0 [=u ee Se te Nes 0 Do. 

11.50 a. m.-12 m ______ 8.5 EE yl RPL De Do. 

12 tna 22 areal nia: PRET Al coy 9s ok 18 | 


Ix, B, 6 Gibson & Concepcion: Lymphagogic Action of Mango 507 


TABLE II].—Haperiment 12, June 4, 1914. Male dog, weighing 9.4 


11.34-11.44 a.m __ 
TY LO CU ee ee See 
OUES5.80's: mi == 227s 
MAD OOiae meet hss ess. 


Injection of 20 cc. of fresh mango 


juice. 


The lymph clots. 


kilograms. 
Lymph 
| Time. ny = ae piced | Remarks. 
- ce. mm Min 
10:04-10:'24) a ;my =. sae ae Op ieeeie se Noe he The lymph clots. 
Oe ae ee ee eee on oo 15 
10.39 a.m 13 
10.39-10.49 a. m_____-_---_-- ie eee 2 AE (apo! 
CN Os Uncen ae ae Re dganpaes 22 | 
WUT RE  yrr Se et 0 a el fe Se Fee Sy (alne ae ee | Injection of 17 cc. of fresh mango 
| juice. 
AO DAE be -Ab ae M2. ook OR is. See ee 
AOlbS 10:58:80 ams == 2 5--—-|\eae- 2 225 YT ee | There seems no explanation for this 
transient rise in blood pressure. 
Mtrot— ht OG to MN 2252888 foie fete er ey Se bee The lymph clots. 
HCAS 7250 ca ERE ee ee leo Ne er 68 
DOS aN eee een Sa Oe oe oe eet 8.5 
Pe Ora mee oe |e es N20 beer ee om | | 
PiGraern seston Me 8 146) [een eee | 
MOWAT as moe =e ee Ce VS Se eS ee + OC uite = a ses Do. 
SUS iets iran ee aD ol eat ee We 11 
ES epee ee ae el eas pha eke ae cee 
ULI UR Ear she I NT ae te ae rH Soc | Injection of 17 cc. of fresh mango 
| | juice. 
11.20.80-11.30.30 a. m__..__-__ 4.5 | BON ie oe The lymph clots. 


MEAG) ASIN eee oa en 


46:80 lal os Be ae 
11.47 a. Ee eee eee 


juice. 


The Philippine Journal 


TABLE I1V.—Experiment 8, May 5, 1914. 


of Science 


Male dog, weighing 9.5 


kilograms. 
Lymph 
| Time. in Bassin cag Mier Remarks. 
ce. mm. Min 
5-10-25 wasn! = Se Ls Feaiig] Reberereh fan CCR gee The lymph clots. 
2G BPN EG +: Rot gee ap ieee es ees; (EL Ne emg nere| UR 10 
0 O1501 Ai eam) ee ee G6 eee eee Do. 
DN-82 as is eo ES ee 14 
14.65 \a. moe ee ee i 53  NAS Seee eee Injection of 25 cc. of fresh mango 
juice. 
PL bGiajm e220 nk es TA ee 
11.56 a. m.-12.06 p. m______- a Hep = ee Rs Ua he The lymph clots. 
RTD Go son eo ee ea soe ote Seer canoe 17.5 
1208 PM eee oe cee ee peal ace eee Lek eee 15 
I2OTI2 AT pom aoe 1G? peso ce REA et eee Do. . 
PLAT LAL) 11 Been Se oe ace semetenens) (aerate. /e Skeet 12 
1219-1228 pea eee eee Sie seeeee coe lessoseene Do. 
12:03 pk ae eS ee oe ee V7.) | ee 
12-31 poms. soo ee eee oN aoe mete pT} ee Bae eS Injection of 25 cc. of fresh mango 
juice. 
ERG ACNE 1 BR ens Sle eel ee re eae a, G4 | seeeee Sec 
nL et VA | 11 ee 1 eae a ee ponnene The lymph clots. 
DUA. MRS 1 is Se ae ee eel eh | ee | 10 
ADI SO in sim re ee ee a eee 70 10 Blood pressure remained low and 


the dog was killed. 


OBSERVATIONS ON MANGO RASH 


By ISABELO CONCEPCION 


(From the Department of Physiology, College of Medicine and Surgery, 
University of the Philippines) 


INTRODUCTION 


It is a common belief among the people of the Philippine Islands 
that the transient rashes of erythematous type occurring during 
the hot season are caused by the eating of mangos (Mangifera 
indica L.). However, there is no definite evidence to indicate 
that the mango is the causative agent. It has long been re- 
cognized that strawberries, crustaceans, and oysters are the 
etiologic factors in certain urticarias and other rashes, local 
cedemas, etc. These substances seem even to be able to exercise 
an influence on the nursing baby through the milk of the 
mother.' 

The object of the present study is to find out whether the 
mango in the Philippines is really a responsible factor for some 
of these transient rashes occurring so commonly from April to 
July. At the same time observations were made to ascertain if 
this fruit has any influence on the nursing baby through the milk 
of the mother. 

Four newly delivered nursing mothers in the obstetrical ward 
of the Philippine General Hospital were chosen and were fed 
usually six mangos daily with their meals. Careful attention 
was given to select patients who were free from skin rashes. 
Daily observations were made on both mother and baby, partic- 
ularly as to the development and disappearance of rashes; sub- 
jective symptoms were noted carefully. Purgatives were given, 
when necessary, to eliminate intestinal auto-intoxication as far 
as possible. It may be stated that both mother and baby were 
sleeping under mosquito nets so that mosquito bites were prac- 
tically excluded. 


*Firmin observed urticaria in an infant whose nursing mother had 
partaken of oysters and fish. Cited from Pfaundler and Schlossman, Dis- 
eases of Children, translated by H. L. N. Shaw and L. La Fetra. J. B. 
Lippincott Company, New York (1908), 4, 443. 

509 


510 The Philippine Journal of Science 1914 


CASE I 


N. C., Filipino woman, 24 years old, married to an American. She was 
admitted to the hospital on June 6, 1914, and delivered on the same day. 
Family history is negative. Previous history is unimportant, except that 
she states that she is prone to have a rash after eating several mangos. 
The patient is fairly well developed and well nourished, and shows no signs 
of dyspnea or discomfort. Examination of the systems is negative. 

The baby (female) was normally delivered, and is fairly well developed. 
This is the fourth child. Subjective examination of the systems is negative. 
The skin is light colored. 

June 6. Four mangos only were given this day. 

June 7. Examination of the mother showed a few papulomacular areas 
with some vesicles. These are of a uniform size, about that of a pinhead 
or a little larger, and are purplish red. These eruptions are located on 
the interscapular region, and itch considerably. Temperature is normal. 
She is constipated. 

Examination of the baby is negative. 

June 8. The mother has developed more areas of the same kind on her 
back. She complains of marked itching. She is constipated. 

The baby is apparently normal. 

June 9. Some vesicular eruptions of about a pinhead in size have 
appeared on both sides of the neck of the mother. She complains of 
marked itching. She was given a cathartic this morning, and had four 
bowel movements. 

The baby shows a few maculopapular eruptions on the cheeks. These 
are about 1 millimeter in diameter. 

June 10. The mother has developed numerous papular vesicles on her 
chest and neck and a few on the extensor surfaces of both arms. The 
temperature is normal. She has had 6 bowel movements. 

The eruptions on the baby’s face have disappeared, but she has developed 
a few vesiculopapular areas on the neck. In the afternoon of the same 
day the rash had appeared abundantly on the neck and to a slight extent 
on the back. The baby is otherwise comfortable. No subjective symptoms 
of any kind were noted. 

June 11. Examination of the mother shows an extensive acute vesiculo- 
papular rash on her chest, back, and neck. Her bowels are normal. Tem- 
perature is normal. She complains of marked itching. 

The baby has developed a few eruptions on the extensor surfaces of the 
lower extremities. Those on the neck and back are still present, 

June 12. In the mother, the rash is not so acute as on the previous day. 
Itching is very much diminished. 

The baby still shows a few areas on the neck, but those found on the 
extremities have almost disappeared. 

June 18. The mango feeding was discontinued. Calomel and saline 
purgative were given to the mother, and she had two bowel movements. 
She no longer complains of itching. Both mother and baby look very much 
as on the previous day. 

June 15. The eruptions on the chest and back of the mother are very 
much diminished, and the vesicles are in a drying condition. 

The baby has developed a few more papulovesicular eruptions on the 
neck. Those found on other parts of the skin have disappeared. 


IX, B, 6 Concepcion: Observations on Mango Rash 511 


June 16. The rash still persists to a slight degree in both mother and 
baby. They were discharged from the hospital subsequent to the 
examination. 

CASE II 


J. L., Filipino woman, 27 years old, married. She was admitted to the 
hospital on June 20, 1914, and delivered on the next day. Family history 
and previous diseases are negative. The patient is fairly well developed, 
pale, and lying flat on bed without any sign of dyspnoea or discomfort. 
Examination of the systems is negative. 

The baby (male) was prematurely born and is small. The skin is dry; 
the subcutaneous tissue is very loose. 

Mango feeding was begun June 21. The mother was given 6 mangos 
for five days, 5 mangos for a day,.and then 8 mangos for a day. She was 
observed twice a day during the period of the experiment. After eight 
days of feeding she failed to develop a rash. 

On the eighth day, the baby had a few papulovesicular eruptions on the 
face which lasted for two days. 

The mother and child were discharged on June 29. 


CASE III 


A. P., Syrian woman, 26 years old, married. She was admitted to the 
hospital on June 20, 1914, and delivered of a female child on the next day. 
Family history and history of previous diseases are negative. Hxamina- 
tion shows some papules on the mammary regions in drying condition. 

June 21. Mango feeding was begun. 

June 22. The mother complains only that her bowels are constipated. 
There is no rash either on the mother or baby. 

June 23. The mother complains of marked itching of the skin of the 
chest and in the axillary regions. On examination, there are many papules 
and papulovesicles in acute condition on the above-mentioned places. 

The baby has developed a few papulovesicles on the face. 

June 24. The mother complains less of itching. The eruptions found 
yesterday are very much diminished in extent. 

The previous rash of the baby is much increased. She has also developed 
some new areas on the extensor surfaces of her legs and thighs. 

June 25. Mango feeding was discontinued. The mother does not com- 
plain of itching. Eruptions on her body have disappeared. 

Examination of the baby shows that of the papules of the previous day 
only very slight traces are left on the face. 

June 26. The mother still has some traces of the rash; otherwise she 
is comfortable. 

The baby is almost the same as on the previous day. 

June 27. The mother is clear from the rash of the previous days. 

The baby is apparently normal. 

June 28. Mother and baby are apparently normal. 

June 29. The mango feeding was resumed. The mother and baby are 
negative. 

June 30. The mother and the baby are apparently normal. 

July 1. The mother is negative. 

The baby has developed a few papules with some papulovesicles on the 
face and also on the extensor surfaces of the arms. 


512 The Philippine Journal of Science 1914 


July 2. The mother is normal. 

The baby has a few eruptions on the arms, although these are not so 
severe as on the previous day. The baby and the mother were discharged 
from the hospital after the examination. 


CASE IV 


P. C., Filipino woman, 20 years old, married. She was admitted to the 
hospital on June 20, 1914, and delivered on the next day. Family history 
and history of previous diseases are negative. The patient is fairly well 
developed, and is well nourished. She has apparently no signs of dyspnea 
nor discomfort, but is not able to be about. Examination of the systems is 
negative. 

The baby (male) was normally delivered, and is fairly well developed. 
Examination of the systems is negative. ” 

June 21 Mango feeding was begun. 

June 22. The mother has developed a few maculopapules on her right 
arm. No itching nor burning sensation is felt. The bowels are con- 
stipated. 

The baby is apparently normal. 

June 23. The mother complains of marked itching. She has developed 
many small papules and vesicles on the neck and breasts. She was given 
a saline purgative, and had seven movements during the day. 

The baby is normal. 

June 24. Examination of the mother shows that the eruptions of the 
mammary region have almost disappeared, but she has developed a few 
of the same kind on her back. She still complains of marked itching. 

The baby shows some papulovesicular rash on the face and neck. 

June 25. Mango feeding was discontinued. The mother does not com- 
plain of itching. The vesicles on her chest are in a drying condition. 

The baby still shows a few areas on the face. 

June 26. The mother is almost free from rash. Itching has discontinued. 

Examination of the baby is negative. 

June 27. Mango feeding was resumed. 

June 28. The mother again complains of slight itching. On examination 
it was found that she has developed a few eruptions of papulovesicular 
character on the neck, shoulder, and infraclavicular regions. 

The baby has also developed a few papulovesicles on the neck. 

June 29. The mother shows some erythema on the neck and back. The 
itching has discontinued. 

The baby still has a few papulovesicles on the face. 

The mother and child were discharged from the hospital subsequent to 
the examination. 


DISCUSSION AND CONCLUSION 


The frequent occurrence of these rashes in a certain season 
of the year, generally from April to July, coincident with the 
hot season, leads us to a suspicion that these rashes are nothing 
more than the ordinary miliaria rubra frequently observed at 
thistime. But when it is considered that these rashes were devel- 
oped even during the cool typhoon periods and that they may be 


IX, B, 6 Concepcion: Observations on Mango Rash 513 


made to appear and disappear by discontinuing and renewing the 
mango feeding, it can be concluded that the etiology of these 
rashes is to be ascribed to the fruit itself. 

The rash observed on the mother after mango eating is of 
the papulovesicular type with small papules and vesicles. These 
are characterized by more or less persistence of the lesions, 
marked itching, and absence of other subjective symptoms. The 
size of the papules and vesicles varies from that of a pinhead to 
about 2 millimeters in diameter. They are round or sometimes 
oval in shape, and appear in patches. The rash must be classed 
as an erythema and not as urticaria. The commonest places for 
the rash to develop are on the mammary regions, the neck, and 
the extensor surfaces of the upper extremities. The eruptions 
found on the baby are generally of the maculopapular type. In 
some instances, the papulovesicular type can also be found. 
They appear usually either singly or in patches. Their shape 
is circular, and they are comparatively larger than those found 
on the mother. The lesions are also less persistent, as compared 
to those in the mother, and are less irritating. They generally 
develop on the face, the neck, and the extensor surfaces of the 
extremities and not infrequntly on the back. 

Other points of interest in connection with these observations 
may be mentioned. In all of the above cases, except one (case 
IJ), rashes were produced in both mother and child; this excep- 
tion indicates that individual susceptibility is a factor in the 
development of the rash. After successive days of feeding, the 
mother may acquire some degree of immunity or tolerance, as 
shown by case III, in which renewal of the mango feeding 
failed to produce a second rash. The rashes have appeared on 
the babies whenever mangos were given to the mothers. The 
babies, therefore, seem to be more susceptible than are the adults. 

In conclusion, I wish to express my thanks to Prof. R. B. Gibson 
for his suggestions and help in carrying out these observations. 
My thanks are also due to Dr. M. Tolentino, of the department of 
obstetrics of the Philippine General Hospital, who secured the 
patients for the experiments. 


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MILK POISONING DUE TO A TYPE OF STAPHYLOCOCCUS ALBUS 
OCCURRING IN THE UDDER OF A HEALTHY COW? 


By M. A. BARBER 
(From the Biological Laboratory, Bureau of Science, Mamila, P. I.) 


During the years 1909-1913 cases of acute gastroenteritis 
occurred repeatedly on a certain farm in Nueva Ecija Prov- 
ince, Luzon, P. I. The symptoms resembled those of so-called 
ptomaine poisoning. Vomiting and diarrhcea were constant 
symptoms, sometimes accompanied by muscular cramps and 
faintness. Patients were usually well enough to be about within 
a few hours, although weakness and malaise often persisted 
a day or so. 

Occasional cases occurred among the American residents of 
the farm and among the Filipino employees, but more often 
visitors were attacked. The origin of the trouble could not be 
found in any kind of food, since this was carefully prepared, 
and consisted of the articles ordinarily used in other households 
where no trouble occurred. Fresh milk and cream, obtained 
from two cows kept on the farm, were in daily use, but these 
were not suspected, since they were freely used by the children 
of the family, and supplied to the family of a neighbor, none 
of whom had any trouble. The water of the well was suspected, 
but chemical and bacteriological analyses, and tests made by 
drinking quantities of samples sent to the laboratory of the 
Bureau of Science, gave wholly negative results. 

The cases showed a curious seasonal incidence, occurring 
almost wholly during the hot dry season of the year—March, 
April, May, and June—and never, or rarely, during the rainy 
or the cool dry season. Few or no cases occurred during the 
year 1911. 

I made several visits of investigation to the farm, and was 
three times the victim of attacks there. No member of the family 
showed any symptoms at the time of these attacks, or at most 
very slight ones, although on two occasions the same articles of 
food and drink were served to all. On one visit, I drank the 


*Read before the Far Eastern Medical Association, December, 1913. 
515 


516 The Philippine Journal of Science 1914 


fresh milk of both cows with no bad results. It was noted, 
however, that all of these attacks followed the ingestion of fresh, 
sweet cream, after an interval of one and three-fourths to 
two and a half hours. So milk was drawn directly into cleaned 
bottles, and brought to the laboratory, keeping it in contact with 
‘ice or in a refrigerator. The milk of the two cows was kept 
separate. Something over twenty-four hours after the milk 
had been drawn, I took a dose of 30 cubic centimeters of the 
cream of one of the cows. This sample had not been above 
refrigerator temperature. No symptoms followed. The bottle 
from which the sample was taken was then left at a room tem- 
perature of 28-30° C. for five hours, and a second dose of 40 cubic 
centimeters of mixed milk and cream was taken. About one 
and three-fourths hours afterwards, a decided nausea and diar- 
rhea set in, which lasted several hours. Symptoms were similar 
to those experienced before, but much milder, and recovery was 
complete within a few hours. For confirmation, several volun- 
teers took doses of the cream of both cows. No symptoms fol- 
lowed the use of the milk and cream from one cow, but three 
persons who took cream from the one tested by me showed 
decided symptoms; in one case, that of a person who took 50 
cubic centimeters, the attack was violent, and the illness lasted 
about two days. 

Agar cultures made from milk, drawn with all precautions into 
sterile test tubes, gave micrococci in both cows. In the suspected 
cow, these were of two sorts. <A yellow staphylococcus from a 
quarter of the udder which, three years previously, had been the 
seat of an attack of garget, and a white staphylococcus occurring 
abundantly in practically pure culture in milk from the other 
quarters. The other cow gave practically a pure culture of a 
yellow staphylococcus differing in color and consistency of col- 
onies from the yellow staphylococcus of the suspected cow. 

Transfers were made from pure cultures of both the yellow 
and the white staphylococci from the suspected cow into flasks 
containing preserved milk from Norway. Both flasks were in- 
cubated at 36°.5 C. for about eight and a half hours. A control 
flask from the same tin, also incubated,.was proved by plate 
cultures to be sterile. A dose of 50 cubic centimeters of the milk 
inoculated with the yellow staphylococcus was taken by me and 
gave no symptoms. A similar dose of 50 cubic centimeters 
from the flask inoculated with the white staphylococcus was 
taken the next day—the flask having remained in the refrigerator 
after removal from the incubator. In one and three-fourths 


ExR 6 Barber: Milk Poisoning 517 


hours an attack of gastroenteritis followed, similar in all details 
to the ones experienced on the farm, but somewhat more violent 
than any of them. The cramps and faintness, especially, were 
pronounced. The more violent symptoms disappeared within 
seven or eight hours, although slight cramps persisted much 
longer. The next day there remained only malaise and a slight 
digestive disturbance. 

The evidence, then, is conclusive that the illness is due to a 
poison formed by the white staphylococcus in milk. The yellow 
staphylococcus, apparently, is unable to produce this toxin. 

The white and the yellow staphylococci from the suspected 
cow were compared on various media, including some 12 different 
litmus agars. The cocci of the white form are somewhat smaller 
than those of the other, and show a greater tendency to form 
diplococci. Cultures further differ in that the white strain 
formed acid on mannite and maltose litmus agars, while the 
yellow did not. Both ferment lactose, but the white in a less 
degree than the yellow. Agglutination tests were made with 
my serum after 5 attacks of the illness and within one 
month after the last attack. The yellow form gave complete 
agglutination in dilutions from 4%, up to %, differing in various 
tests, while the white never gave it in dilutions higher than 45. 
In a test with the serum of a resident of the farm, who was 
tolerant of the poison, the white form failed to give complete 
agglutination in a dilution higher than 4. 

Inoculated subcutaneously into guinea pigs and monkeys, the 
white form gave decidedly more reaction than the yellow—in 
- some cases abscesses were formed by it, which later healed. 
Rabbits have resisted doses of the white variety up to 0.3 cubic 
centimeter of an 18-hour broth culture, given intravenously. 
Kittens, pups, and monkeys show no symptoms on ingesting 
large quantities of milk cultures of the white staphylococcus; or, 
at all events, the symptoms are very slight. A goat fed with 
3 slants of the culture showed no cocci in the milk. 

The cow from which the toxin-producing staphylococcus was 
obtained is of mixed breed. She was imported from Australia, 
and came into the possession of the present owner in 1909. . She 
has been immunized to Texas fever and has had rinderpest and 
foot and mouth disease, all previous to 1909. In 1910 she had 
garget in one quarter of the udder, but has been apparently 
healthy since. It is noteworthy that the quarter which suffered 
from garget has shown few or none of the toxin-producing sta- 
phylococci, while this organism occurred in nearly pure culture 


518 The Philippine Journal of Science 1914 


in the other three quarters. Since several cases of the poisoning 
were noted previous to the occurrence of garget in the cow, this 
could not have been the origin of the trouble. A young cow, 
the offspring of the infected one, has been in contact with the 
mother about two years, and gives apparently wholesome milk. 

Staphylococci are commonly found in the milk of apparently 
healthy cows, but I have been unable to find in the litera- 
ture any reference to the formation by a staphylococcus of a 
toxin producing the symptoms above described. The staphy- 
lococcus isolated in this case may be an unusual strain; or, it is 
possible that it is not uncommon, but that the usual practice 
of keeping milk refrigerated prevents the formation of the 
toxin. Ice was not in daily use on this farm, and as stated above 
cream from the infected cow which had been kept cold for 
twenty-four hours did not give the symptoms, but these followed 
the ingestion of cream from the same bottle after it had been 
left five hours at room temperature. As noted above, the cases 
of gastroenteritis occurred chiefly during the hot months of the 
year. This was the season when the milk from the infected cow 
was mostly used, but the attacks at this season may have been due 
in part to the higher temperature which favored the growth 
of the staphylococcus. There was nothing unusual in the taste 
of the infected cream. 

Since the cause of the poisoning became known, the use of 
the raw milk of the infected cow has been discontinued, and 
all trouble has ceased. One person during a residence of several 
months on the farm had suffered from indigestion of a chronic 
character, but had had few acute attacks. With the discontin- 
uance of the use of infected milk his indigestion ceased. 

As has been stated, milk from the two cows was regularly 
furnished to a neighbor, and no attacks occurred in his family. 
His freedom from the trouble is probably due to the fact that 
the milk was commonly used fresh in his household. In the 
cases of the family of the owner of the cow, some resistance 
to the toxin had apparently been developed. On two occasions 
at least when I suffered attacks at the farm, the infected cream 
was eaten by members of the family who subsequently showed 
no decided symptoms. A sample of the cream which had caused 
a severe attack in me was preserved, and the next day a member 
of the family took a large dose of it. Only slight symptoms 
followed. The cream had become sour in the meantime. The 
evidence that all members of the family had acquired some toler- 
ance to the toxin seems conclusive. 


IX, B, 6 Barber: Milk Poisoning 519 


In summary, the most noteworthy points of this paper are the 
following: 

Acute attacks of gastroenteritis were produced in milk by a 
toxin elaborated by a white staphylococcus which occurred in 
almost pure culture in the udder of a cow. The fresh milk was 
harmless, and the toxin was produced in effective quantities only 
after the milk had stood some hours at room temperature. 

Repeated attacks of the illness had occurred among residents 
and visitors at the farm during a period of three years, and 
the cow was apparently in good health during this time, except 
for one attack of garget, which occurred after the cases of gastro- 
enteritis had begun. 

Persons who had used the milk continuously had apparently 
developed some tolerance to the toxin. Two children of the 
family had used the milk regularly, but never had attacks. The 
adults had occasional light attacks or, in one or two cases, some 
chronic intestinal trouble. In my own case, 4 acute attacks, 
3 of them severe, afforded no protection against a subsequent 
fifth dose. Visitors at the farm and Filipino employees who 
used the milk less regularly showed most severe attacks. Since 
the discontinuance of the use of raw milk from this cow, all 
trouble has ceased. 

Culturally, the toxin-producing staphylococcus differed little 
from a nontoxin-producing strain, except that the former pro- 
duced acid in mannite and maltose litmus agars. 

Agglutination tests with the serum of a person who had 
recently suffered 5 attacks and that of a person who had long 
used the milk showed little, if any, positive result. 

In guinea pigs and monkeys the toxin-producing strain showed 
more tendency to form abscesses than a yellow staphylococcus 
from the same source. 

Cases of gastroenteritis occurring in the tropics and in the 
warm season elsewhere may be due to a toxin of similar origin, 
especially where fresh milk is not properly refrigerated before 
use. This is the more probable since staphylococci of various 
types commonly occur in the udders of apparently healthy cows. 

ADDENDUM.—The cow which harbored the toxin-producing 
Staphylococcus was sold to the College of Agriculture at Munoz, 
Nueva Ecija. A letter from the superintendent of the school, 
Mr. Kilmer O. Moe, dated January 6, 1915, states that the milk 
of the cow is being used by the students at present. However, 
it is used fresh and principally in coffee; no bad results have 
followed its use. 


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REVIEWS 


Morris’s | Human Anatomy | a complete systematic treatise | by English and . 
American authors | edited by | C. M. Jackson, M. S., M. D. | professor 
and director of the department of anatomy, | University of Minnesota | 
eleven hundred and eighty two illustrations | three hundred and fifty 
eight printed in colours | fifth edition, | revised and largely rewritten | 
Philadelphia | P. Blakiston’s Son & Co. | 1012 Walnut Street | No 
date; copyright, 1914. Cloth, pp. i-xiv-+-1-1539. Price, $6. 


This book represents the combined efforts of twelve of the 
most prominent American and English anatomists, who have 
succeeded well in their attempt to revise and revive a textbook 
in anatomy which was rapidly falling into disuse. The editor 
has put forth special effort to discriminate systematically be- 
tween fundamental and detailed anatomical facts. To meet 
these conditions, the book is printed in a large and small type; 
the former is used to indicate the fundamental facts, the latter, 
the details of anatomy. This arrangement should meet the ap- 
proval of the student. 

As previously indicated, the book contains a large number of 
illustrations, almost one-third of them in colors. It appears as 
though special emphasis and stress has been placed on quantity 
rather than quality, as some of the prints are very indistinct 
and of little value to the student. Some of the prints are new, 
but most of them are taken from previous editions and other 
textbooks. 

The Anglicized form of the BNA is used with a few excep- 
tions where the original BNA or the Latin form is adopted in 
the English. Since the general adoption of the BNA, anatomical 
terms have been reduced from 30,000 to 5,000. It seems-obvious, 
therefore, that with the large reduction in the number of terms 
the student could and should master the Latin anatomical ter- 
minology adopted by the International Congress at Basel. While 
the Anglicized form, in most cases, differs but little as a whole 
from the original BNA, yet in some instances the terms are 
vastly different, and scarcely recognized by the student. 

The last section on clinical and topographical anatomy could 
well have been omitted, as they are rarely if ever referred to 
in the study of systematic anatomy. 

521 


522 The Philippine Journal of Science 


Both clinical and topographical anatomy should be taught as 
individual courses, and the section dealing with them is entirely 
too inadequate for a student’s guide. 

As a whole the book is reliable throughout and can be safely 
recommended as one of the standard textbooks in anatomy. 

E. S. RuTH. 


The Diagnosis and Treatment | of | Tropical Diseases | by | E. R. Stitt, A. B., 
Ph. D., M. D. | medical director, U. S. Navy; [etc., 8 lines] | with 
86 illustrations | Philadelphia | P. Blakiston’s Son & Co. | 1012 
Walnut Street | No date; copyright, 1914. Cloth, round corners, pp. 
i-xi+1-421. Price, $2. 

This little volume should commend itself to medical men gen- 
erally as a clear and concise exposition of what is known of 
tropical diseases. Despite numerous typographical errors the 
book can be heartily commended. 

J. A.J. 


INDEX 


A 


ACOSTA-SISON, HONORIA, Pelvimetry and 
cephalometry among Filipinas, 493. 
Amebic and bacillary dysenteries 
Philippine Islands, data on, 205. 
Anopheles albimanus, 427. 

barbirostris van de Wulp, 384, 389. 

(Celia) albimanus, 434. 

eulicifacies, 387. 

febrifer Banks, 384. 

fuliginosus, 391. 

funesta, 384. 

jesoensis Tsuzuki, 389. 

karwari, 390, 391. 

larve in captivity, development of, 
421. 

ludiowii Theobald, 384, 385. 

maculata Theobald, 390. 

maculatus Theobald, 384, 390. 

malefactor, nontransmission of ma- 
laria by, 381. 

(Myzomyia) febrifer sp. nov., the 
transmission of malaria with, 381 
et seq. 

(Myzorhynchus) barbirostris, the 
transmission of 
malaria with, 
381 et sea. 

sinensis, the 
transmission of 
malaria with, 
881 et seq. 

(Nyssorhynchus) fuliginosus Giles, 

391. 

maculatus, the 
transmission of 
malaria with, 
381 et sea. 

(Pseudomyzomyia) rossii, the 
transmission of malaria with, 
381 et seq. 

pseudopunctipennis, 427. 

rossii Giles, 384, 385. 

sinensis Wiedemann, 384, 389. 

(jesoensis), 390. 
vagus Donitz, 385. 
“Anopheles I,”’ 387. 
“Anopheles Ia,”’ 391. 
“Anopheles II,” 389. 
Anophelines, avidity of, for human blood, 424. 
Anthrax bacillus, 466 et sea. 
Ants as carriers of Asiatic cholera, 
roaches and, 1. 
Apparatus, some simple laboratory, 297. 
venous-pressure, 297. 


in the 


cock- 


Ascaris, 81, 83, 178, 234 et seq., 446 et seq. 
Asiatic cholera, notes on the diagnosis of, 361. 
Aurellia labiata Cham. and Eysen., 294. 
Autopsies, the chief intestinal lesions in one 
thousand, 453. 
B 


Bacillary and ameebie dysenteries in the Phil- 
ippine Islands, data on, 205. 
colitis, 458, 457 et sea. 
dysentery: The most prevalent form 
in Manila and its treatment, 241. 
Bacillus coli, 244, 
dysenteriz, 242 et seq. 
lepre, 227, 230. 
pyocyaneus, 347. 
“S,” 244 et seq. 
tuberculosis, 338, 340. 
typhosus, 253 et sea. 
Bacillus of anthrax, 466 et seq. 
diphtheria, 466 et seq. 
glanders, 466 et sea. 
plague, 466 et seq. 
Bacterium coli, 320, 340. 
commune, 326, 329. 
Balantidium, 81, 88, 173, 446 et seq. 
coli, 87, 173. 

BANKS, CHARLES S., Sanitary survey in 
Mindoro. 5. The mosquito survey in and 
near San Jose, 1638. 

BARBER, M. A., Cockroaches and ants as 
carriers of the vibrios of Asiatic cholera, 1; 
The pipette method in the isolation of single 
microérganisms and in the inoculation of 
substances into living cells, 307; review of 
Manson’s Tropical Diseases, a Manual of 
the Diseases of Warm Climates, 463; review 
of Henson’s Malaria Etiology, Pathology, 
Diagnosis, Prophylaxis, and Treatment, 
463; Milk poisoning due to a type of Sta- 
phylococcus albus oceurring in the udder 
of a healthy cow, 515; see also WALKER, 
ERNEST LINwoop, 381. 

Beriberi, in the Philippine Islands, 203. 

rice in relation to, 119. 

Book reviews, see Reviews (book). 

BOYNTON, WILLIAM HUTCHINS, A pre- 
liminary report of experiments on the culti- 
vation of the virus of rinderpest in vitro, 
39; An atypical case of rinderpest in a 
carabao, 45; Experiments on the cultivation 
of rinderpest virus as described by Baldrey, 
259; Kidney-worm infestation of swine in 
the Philippine Islands with special refer- 
ence to the pathological changes, 269; see 
also WARD, ARCHIBALD R., 49. 


523 


524 


Cc 


CALDERON, FERNANDO, Some data con- 
cerning the medical geography of the Phil- 
ippines, 199. 

Carabao, an atypical case of rinderpest in 
a, 45. 

Carabaos, simultaneous method of inoculating, 
126. 

Carus, Paul, see Reviews (book). 

Catostylus purpurus Mayer, 293, 294. 

Cats, plague in, 10. 

Cattle and carabaos, simultaneous method of 
inoculating, 125. 

Celia kochi Dénitz, 384. 

Cephalometry, among Filipinas, 493. 

Cestode, 81, 83. 

Chemiotaxis, experiments on, 341. 

Chiropsalmus quadrigatus Haeckel, 291, 293. 

Cholangitis, 459. 

Cholera, Asiatic, at autopsy, 453. 

cockroaches and ants as car- 


riers of, 1. 
notes on the diagnosis of, 
361. 


spirillum, 466 et seq. 

vibrio in Manila waters, vitality of 
the, 479. 

vibrios, longevity of, 3. 

“Chrysaora state,’”’ 292. 

Clonorchis, 286, 240. 

Cockroaches and ants as carriers of Asiatic 
cholera, 1. 

Colitis, 459. 

CONCEPCION, ISABELO, Observations on 
mango rash, 509; see also GIBSON, R. B., 
119; 503; see also WALKER, E. L., 167. 

Corynebacterium, 220, 222, 224. 

Councilman, W. T., see Reviews (book). 

COX, C. E., review of Reed’s Marriage and 
Genetics, Laws of Human Breeding and 
Applied Eugenics, 463. 

COX, SETH L., see Muscrave, W. E., 178; 
see WALKER, E. L., 181. 

Cropper, J. W., see Ross, H. C. 

CROWELL, B. C., review of Dorland’s The 
American Illustrated Medical Dictionary, 
304; Notes on the diagnosis of Asiatic 
cholera at autopsy, 361; review of Vedder's 
Beriberi, 380; The chief intestinal lesions 
encountered in one thousand consecutive 
autopsies in Manila, 453; review of Lee’s 
Microtomists’ Vade-mecum, 461; review of 
Councilman’s Disease and Its Causes, 462; 
review of Lewis’s A Text-book of Histology 
Arranged upon an Embryological Basis, 462. 

Culex fatigans, 483. 

ludlowii, 164. 


D 


Dactylometra quinquecirrha L. Agassiz, 292 
et seq. 

DALBURG, F. A., Sanitary survey of Min- 
doro. 2. Geography and geology, 142. 

De la Mettrie, Julien Offray, see Reviews 
(book) . 


Index 


Dibothriocephalus, 178. 
latus, 173. 

Diphtheria bacillus, 466 et seq. 

Dissection, technique for, 307. 

Dorland, W. A. Newman, see Reviews (book). 

DUMEZ, A. G., review of Pittenger’s Bio- 
chemic Drug Assay Methods, with Special 
Reference to the Pharmaco-dynamic Stand- 
ardization of Drugs, 379. 

Duodenal ulcer, 453, 458. 

Dysenteries, amcebic and bacillary, in the 
Philippine Islands, data on, 205. 

Dysentery, bacillary, the most prevalent form 
in Manila and its treatment, 241. 


E 


Emetine, treatment of entamebiasis with, 
ipecac, and neosalvarsan, 98. 

Entameba, 178, 446. 

coli, 81 et seq., 93 et seq. 
histolytica, 81 et seq., 93 et sea. 

“Entamoeba tetragena,’’ 97. 

Entamebiasis, in patients of the Philippine 
General Hospital, Manila, P. I., 81; pre- 
liminary report on the treatment of, with 
ipecac, emetine, and neosalvarsan at the 
Philippine General Hospital, Manila, P. I., 
93. 

Entameebic colitis, 458, 455. et seq. 

Enteritis, 459. 

Enterocolitis, 459. 


iy 


Filaria bancrofti Cobbold, 483, 488 et seq. 
demarquayi, 488 et seq. 
magalhaesi, 488 et seq. 
nocturna, 202. 
ozzardi, 488 et seq. 
perstans, 488 et seq. 
philippinensis, 483. 
(Loa) loa, 488 et seq. 
found in the Philippine Islands, 205, 
483. 
Qilipinas, pelvimetry and cephalometry among, 
493. 
Fleas in relation to plague, 12. 


G 


Geography, of the San Jose Estate, 142. 
medical, of the P. L., 199. 

Geology of the San Jose Estate, 142. 

Germicidal power of glycerin on various 
Microérganisms under various conditions, 
465. 

GIBSON, R. B., Some simple laboratory ap- 
paratus, 297; review of de la Mettrie’s Man 
a Machine, 304; review of Howell’s A Text- 
book of Physiology for Medical Students 
and Physicians, 461; A note on the physio- 
logical action of the proteoses, 499. 

GIBSON, R. B., and CONCEPCION, ISA- 
BELO, Nerve degeneration in fowls fed on 
unhusked rice (palay), 119; The lympha- 
gogic action of the Philippine mango, Man- 
gifera indica Linnzus, 5603. 

Glanders bacillus, 466 et seq. 


Index 


Glossina morsitans, 382. 
Glycerin, germicidal power of, 
microérganisms, 465. 
GUTIERREZ, PERPETUO, Typhoid fever in 
the Philippines, 367; see also WILLETS, 
Davin G., 98; see also MuscrAve, W. E., 178. 
GUZMAN, A. M., see WALKER, E. L., 167. 


age 


HEISER, VICTOR G., Reappearance of 
plague in the Philippines after an absence 
of six years, 5. 

Helminthiasis, intestinal, in the Philippine 
Islands as indicated by examinations of 
prisoners upon admission to Bilibid Prison, 
Manila, P. I., 283. d 

Henson, Graham E., see Reviews (book). 

Hering, Ewald, see Reviews (book). 

Hookworm, 81, 83, 173, 179, 236, 446. 

Howell, William H., see Reviews (book). 

Hymenolepis nana, 236. 


I 


on various 


Infection, genitourinary, 459. 

Inoculation, simultaneous method of, of cattle 
and carabaos with serum from animals that 
have been recently immunized, 125. 

Insects as carriers of Asiatic cholera, 1. 

Intestinal helminthiasis in the Philippine 

Islands as indicated by examina- 
tions of prisoners upon admis- 
sion to Bilibid Prison, Manila, 
P. 1., 233. 

lesions encountered in one thousand 
consecutive autopsies in Manila, 
453. 

parasitism, particularly entameebia- 
sis, in patients of the Philippine 
General Hospital, Manila, P. I., 
81. 

Ipecac, treatment of entamebiasis with, 93. 

Isolation of microérganisms by the pipette 
method, 307. 

J 


Jackson, C. M., see Reviews (book). 
JACKSON, THOMAS W., Sanitary survey 


in Mindoro. 4. The internal sanitary prob- 
lem, 149. 

Jellyfish, another dangerous, in Philippine 
waters, 291. 


JOHNSTON, JOHN A., A contribution to 
the bacteriology of leprosy. Preliminary 
note, 227; review of MacNeal’s Pathogenic 
Micro-organisms. A Text-book of Micro- 
biology for Physicians and Students, 303; 
review of Jordan’s A Text-book of General 
Bacteriology, 461; review of Stitt’s The 
Diagnosis and Treatment of Tropical Di- 
seases, 522. 

Jordan, Edwin O., see Reviews (book). 


K 


Kidney-worm infestation of swine in the Phil- 
ippine Islands with special reference to the 
pathological changes, 269. 


525 


L 


Laboratory apparatus, some simple, 297. 
used in the pipette 
method, 307. 
Lamblia, 173. 
Lard worm of pigs, 269. 
Lee, Arthur Bolles, see Reviews (book). 
Leishmania, 167, 169. 
Leprosy, 2 contribution to the bacteriology 
of, 227. 
data on, in the Philippine Islands, 
204. 
Lesions, noninfectious or unclassified inflam- 
matory, of intestines, 458, 458. 
Lewis, Frederick T., see Reviews (book). 
LIGHT, S. F., Another dangerous jellyfish in 
Philippine waters, 291. 
Lobonema mayeri Light, 298, 294. 
smithii Mayer, 2938, 294, 295. 


M 


MacNeal, Ward J., see Reviews (book). 
Malaria, experiments on the transmission of, 
381. 
in the Philippine General Hospital, 
Manila, P. I., during the fiscal year 
1913, 441. 
in the Philippine Islands, 459. 
sanitary survey of the San Jose 
estate, Mindoro, with special ref- 
erence to, 137. 

Malarial fever in the Philippine Islands, data 
on, 203. 

Mangifera indica L., 509. 

Linneus, the lymphagogic | 
action of, 503. 

Mango, Mangifera indica Linnzus, the lym- 
phagogic action of, 503. 
rash, observations on, 509. 

Manson, Patrick, see Reviews (book). 

Medical geography of the Philippines, 199. 

Metastrongylus, 276. 

Microfilaria bancrofti, 202. 

Microérganisms, germicidal power of glycerin 

on, 465. 
the pipette method 
isolation of, 307. 

Milk poisoning due to a type of Staphylo- 
coccus albus in the udder of a healthy cow, 
515. 

Monad, 81, 83. 

Monomorium latinode Mayr as a possible car- 
rier of Asiatic cholera, 4. 

Morphology of the adults of the filaria found 
in the Philippine Islands, 483. 

Mosquitoes, collecting larve of, 392. 

dissection of, 398. 
infecting, 397. 
rearing, 394. 

MUSGRAVE, W. E., Sanitary survey in Min- 
doro. 1. Introduction, 138; 3. The general 
and external sanitary problem, 143. 

MUSGRAVE, W. E., and SISON, A. G., Ba- 
cillary dysentery: The most prevalent form 
in Manila and its treatment, 241. 


in the 


526 


MUSGRAVE, W. E., VAZQUEZ A., GUTIE- 
RREZ, B., and COX, SETH L., Sanitary 
survey in Mindoro. 6. The disease index. 
(B) Clinical examinations, 173. 

MUSGRAVE, W. E., et al., Sanitary survey of 
the San Jose Estate and adjacent proper- 
ties on Mindoro Island, Philippine Islands, 
with special reference to the epidemiology 
of malaria, 137. 

Myzomyia christophersi, 439. 

funesta Giles, 384. 

listoni, 439. 

ludlowii Theobald, 385. 

mangyana Banks, 384. 

rossii Theobald, 164, 385. 

thorntonii Ludlow, 384. 

Myzorhynchus barbirostris Theobald, 389. 

barbirostris van de Wulp, 384. 
peditaeniatus Leicester, 889. 
pseudobarbirostris Ludlow, 384. 
sinensis Theobald, 389. 
sinensis Wiedemann, 384. 
vanus Walker, 384. 


N 


Neosalvarsan, treatment of entamebiasis with, 
93. 
Nephritis, chronic, 459. 
Nerve degeneration in fowls fed on unhusked 
rice (palay), 119. 
Nitella, 356. 
Nyssomyzomyia ludlowii 
385. 
rossii James and Liston, 385. 
Nyssorhynchus fuliginosus Giles, 384. 
maculatus Theobald, 390. 
philippinensis Ludlow, 384. 
pseudowillmori Theobald, 390. 
willmori Leicester nec James, 
390. 


oO 


Opisthorchis (Clonorchis), 240. 
Oxyuris, 81, 83, 173, 235, 446. 


ee 


Palay (unhusked rice), nerve degeneration in 
fowls fed on, 119. 

Paramecium, 356. 

Parasitism, intestinal, particularly entamc- 
biasis, in patients of the Philippine General 
Hospital, Manila, P. I., 81. 

Pelvimetry and cephalometry among Filipinas, 
493. 

Perforation of gall bladder, 459. 

Periplaneta americana Linnzeus as a carrier 
of Asiatic cholera, 1. 

Physiological action of the proteoses, 499. 

Pipette method in the isolation of single mi- 
croérganisms and in the inoculation of sub- 
stances into living cells, 307. 

Pittenger, Paul S., see Reviews (book). 

Plague among rats and other animals, 9, 10. 

bacillus, 466. 
in the Philippine Islands, 5, 19. 


James and Liston, 


Index 


Plague sanitary measures, 14. 
transmission by fleas, 14. 
Plasmodium malariz, 168, 433. 
precox (falciparum), 
432, 483. 
vivax, 168, 432. 
Pneumonia, lobar, 459. 
Poisoning, milk, due to a type of Staphylo- 
coccus albus, 515. 
Polyneuritis in fowls, 119. 
Pottenger, Francis Marion, see Reviews (book) . 
Proteoses, a note on the physiological action 
of the, 499. 
Pseudomyzomyia indefinata Ludlow, 384. 
ludlowii Theobald, 384. 
rossii Giles, 384. 
rossii Theobald, 385. 
Pulex cheopis, in relation to plague, 14. 
Pyretophorus freere Banks, 384. 
minimus Theobald, 384. 
philippinensis Ludlow, 384. 
pitchfordii Giles, 384. 


168, 396, 


R 


Rats in human plague outbreak, 7, 9. 
Reed, Charles A. L., see Reviews (book). 
REVIEWS (BOOK): 

Carus, Paul, The Mechanistic Principle 
and the Non-mechanical. An Inquiry 
into Fundamentals with Extracts from 
Representatives of Hither Side, 380. 

Councilman, W. T., Disease and Its 
Causes, 462. 

De la Mettrie, 
Machine, 3804. 

Dorland, W. A. Newman, The American 
Tilustrated Medical Dictionary, 304. 

Henson, Graham E., Malaria Etiology, 
Pathology, Diagnosis, Prophylaxis, and 
Treatment, 463. 

Hering, Ewald, Memory Lectures on the 
Specific Energies of the Nervous Sys- 
tem, 380. ‘ 

Howell, William H., A Text-book of Phy- 
siology for Medical Students and Physi- 
cians, 461. 

Jackson, C. M., Morris’s Human Anatomy, 
a Complete Systematic Treatise by 
English and American Authors, 521. 

Jordan, Edwin O., A Text-book of General 
Bacteriology, 461. 

Lee, Arthur Bolles, 
Vade-mecum, 461. 

Lewis, Dr. Frederick T., A Text-book of 
Histology Arranged upon an Embryo- 
logical Basis, 462. 

MacNeal, Ward J., Pathogenic Micro- 
organisms. A Text-book of Microbi- 
logy for Physicians and Students of 
Medicine, 303. 

Manson, Patrick, Tropical Diseases, a 
Manual of the Diseases of Warm 
Climates, 463. 


Julien Offray, Man a 


The Microtomists’ 


Index 


REVIEWS (BOOK)—Continued. 

Pittenger, Paul S., Biochemic Drug Assay 
Methods, with Special Reference to the 
Pharmaco-dyanamic Standardization of 
Drugs, 379. 

Pottenger, Francis Marion, Muscle Spasm 
and Degeneration in Intrathoracic In- 
flammations, 464. 

Reed, Charles A. L., Marriage and Gene- 
tics, Laws of Human Breeding and Ap- 
plied Eugenics, 463. 

Robinson, Victor, An Essay on Hasheesh, 
including Observations and lExperi- 
ments, 305. 

Ross, H. C., Cropper, J. W., Ross, E. H., 
Further Researches into Induced Cell- 
reproduction and Cancer, 464. 

Satterthwaite, Thomas E., Cardio-vascular 
Diseases. Recent Advances in their 
Anatomy, Physiology, Pathology, Diag- 
nosis and Treatment, 305, 

Stitt, E. R., Practical Bacteriology, Blood 
Work, and Animal Panasitology, includ- 
ing Bacteriological Keys, Zodlogical 
Tables and Explanatory Clinical Notes, 
304. 

Stitt, E. R., The Diagnosis and Treatment 
of Tropical Diseases, 522. 

Tyson, James and Fussell, Howard, The 
Practice of Medicine. A Text-book for 
Practitioners and Students with Special 
Reference to Diagnosis and Treatment, 
803. 

Vedder, Edward B., Beriberi, 380. 

Rice (palay), unhusked, nerve degeneration in 
fowls fed on, 119. 
Rinderpest, an atypical case of, in a carabao, 
45. 
.a preliminary report of experi- 
ments on the cultivation of the 
virus of, in vitro, 39. 
experiments upon the transmis- 
sion of, 49. 
virus of, experiments on the cul- 
tivation of, as described by 
Baldrey, 259. 
Robinson, Victor, see Reviews (book). 
Ross, E. H., see Ross, H. C. 
Ross, H. C., see Reviews (book). 
RUEDIGER, E. H., The germicidal power of 
glycerin on various microdrganisms under 
various conditions, 465. 
RUTH, E. S., review of Jackson’s Morris’s 
Human Anatomy, 522. 


Ss 


Sanitary survey of the San Jose estate and 
adjacent properties on Mindoro Island, Phil- 
ippine Islands, with special reference to the 
epidemiology of malaria, 137. 

San Jose Estate, sanitary survey of the, with 
special reference to malaria, 137. 

Sarcina, 222. 

Satterthwaite, Thomas E., see Reviews (book). 


527 


Schistosoma japonicum, 236. 

SCHOBL, OTTO, The etiology of trichomyco- 
sis palmellina in the Philippine Islands, 219; 
The vitality of the cholera vibrio in Manila 
waters, 479. 

Sciatic nerve, degeneration of, in polyneuritis, 
122. 

Sclerostoma equinum, 283. 

pinguicola Verrill, 269. 

Scyphomedusz, 294. 

Seatworm, 235. 

Sepsis, 459. 

Serum from animals that have been recently 
immunized, simultaneous method of inocu- 
lating cattle and carabaos with, 125. 

SISON, A. G., review of Tyson and Fussell’s 
The Practice of Medicine. A Text-book for 
Practitioners and Students with Special 
Reference to Diagnosis and Treatment, 303 ; 
see also MUSGRAVE, W. E., 241. 

Smallpox in the Philippine Islands, data on, 
204. 

Spirillum of cholera, 466 et seq. 

Spirogyra, 344, 356. 

Staining, technique for, 307. 

STANLEY, COURT R., see WILLETS, DAviID 
G., 93. 

Staphylococcus albus, 465 et seq. 

albus, milk poisoning due to a 
type of, 615. 
aureus, 222, 466 et seq. 
Stegomyia, 356. 
calopus, 436. 
pseudoscutellaris, 489. 

Stephanurus dentatus Diesing, 269 et sea. 

Stethomyia pallida Ludlow, 384. 

Stitt, E. R., see Reviews (book). 

Streptothrix, 227. 

Strongyloides, 81, 83, 173, 236 et seq., 446 et 

seq. 
stercoralis, 173. 
Strongylus armatus, 276, 283. 
Swine, kidney-worm infestation of, 269. 


dk 


Tenia, 236 et seq. 
Thermoesthesiometer, 297. 
Timing system, a simple, 298. 
Transmission of rinderpest, 49. 
Trauma, 459. 
Trichomonas, 173. 
Trichomycosis palrmellina, 

Philippine Islands, 219. 
Trichuris, 81, 88, 178, 233 et seq., 446 et seq. 
Trypanosoma rhodesiense, 382. 
Tuberculosis, data on, in the Philippine Is- 

lands, 206. 
intestinal, 453 et seq. 
Typhoid bacillus, 465 et seq. 
fever in the Philippine Islands, 206, 
367, 453, 455. 

Tyson, James, and Fussell, Howard, see Re- 

views (book). 


etiology of, in the 


528 


U 

Uremia, 459. 

Vv 

Vaucheria, 356. 

VAZQUEZ, A., see MuscRAvE, W. E., 173. 

Vedder, Edward B., see Reviews (book). 

Vibrio, vitality of the cholera, in Manila 

waters, 479. 
Vibrios, cockroaches and ants as carriers of 
the, of Asiatic cholera, 1. 

Virus of rinderpest, a preliminary report of 
experiments on the cul- 
tivation of the, in vitro, 
39. 

experiments on the culti- 
vation of, as described 
by Baldrey, 259. 


Ww 


WALKER, ERNEST LINWOOD, The mor- 
phology of the adults of the filaria found in 
the Philippine Islands, 483. 

WALKER, ERNEST LINWOOD, and BAR- 
BER, MARSHALL A., Malaria in the 
Philippine Islands, I. Experiments on 
the transmission of malaria with Ano- 
pheles (Myzomyia) febrifer sp. nov., Ano- 
pheles (Pseudomyzomyia) rossii, Anopheles 
(Myzorhynchus) barbirostris, Anopheles 
(Myzorhynchus) sinensis, and Anopheles 
(Nyssorlhynchus) maculatus, 381. 

WALKER, E. L., and COX, S. L., Sanitary 
survey of Mindoro. 7. A comparison of the 
spleen index with the microscopic examina- 
tion of the blood for malarial parasites in 
1,003 persons, 180. 


Index 


WALKER, E. L., GUZMAN, A. M., and 
CONCEPCION, L., Sanitary survey in Min- 
doro. 6. The disease index. (A) Labora- 
tory examinations, 167. 

WARD, ARCHIBALD R., and WOOD, FRE- 
DERICK WILLAN, Simultaneous method of 
inoculating cattle and carabaos with serum 
from animals that have been recently im- 
munized, 125. 

WARD, ARCHIBALD R., WOOD, FRED- 
ERICK WILLAN, and BOYNTON, WIL- 
LIAM HUTCHINS, Experiments upon the 
transmission of rinderpest, 49. 

Waters, vitality of the cholera vibrio in Ma- 
nila, 479. 

Widal reactions among healthy adult Filipinos, 
253. 

WILLETS, DAVID G., Intestinal parasitism, 
particularly entamebiasis, in patients of 
the Philippine General Hospital, Manila, 
P. I., 81; Preliminary report on the treat- 
ment of entameebiasis with ipecac, emetine, 
and neosalvarsan at the Philippine General 
Hospital, Manila, P. I., 98; Intestinal helmin- 
thiasis in the Philippine Islands as indicated 
by examinations of prisoners upon admission 
to Bilibid Prison, Manila, P. I., 238; Widal 
reactions among healthy adult Filipinos, 
268; review of Stitt’s Practical Bacteriology, 
Blood Work, and Animal Parasitology, in- 
eluding Bacteriological Keys, Zodlogical 
Tables, and Explanatory Clinical Notes, 304; 
Malaria in the Philippine General Hospital, 
Manila, P. I., during the fiscal year 1918, 
441. 

WOOD, FREDERICK WILLAN, see Warp, 
ARCHIBALD R., 49, 125. 


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gether with the results of certain bacte- 
rlological investigations, constitute the pres- 
ent report. 

The Bureau. of Science of the Govern- 


ment of the Philippine Islands has been 


appointed sole agent for the distribution 
of the printed proceedings of the Interna- 
tional Plague Conference, 


PRICES ARE IN UNITED STATES CURRENCY 


Orders for these publications may be sent to the BUSINESS MANAGER, 
PHILIPPINE JOURNAL OF SCIENCE, BUREAU OF SCIENCE, MANILA, P. I,, 


or to any of the agents listed below. 


Please’ give order number. 


The Macmillan Company, 64-66 Fifth Avenue, New York, U. S. A. 
Wm. Wesley & Son, 28 Essex Street, Strand, London, W. C., England. 
Martinus Nijhoff, Lange Voorhout 9, The Hague, Holland. 

Mayer & Miller, Prinz Louis Ferdinandstrasse 2, Berlin, N. W., Germany. 
Kelly & Walsh, Ltd., 32 Raffles Place, Singapore, Straits Settlements. 
A. M. & J. Ferguson, 19 Baillie Street, Colombo, Ceylon. 

Thacker, Spink & Co., P. O. Box 54, Calcutta, India. 


Grae wow eA 
BA PEE ge 


CONTENTS 
. Page 
RUEDIGER, E. H. The Germicidal Power of Glycerin on 
Various Microédrganisms under Various. Conditions............- 465 
SCHOBL, OTTO. The Vitality of the Cholera Vibrio in Manila — 
1g) ey RN eS ane BILLS an sa ER fl Se a a CE 479 
WALKER, ERNEST LINWOOD. The Morphology of the Adults 
of the Filaria Found in the Philippine Islands... 2.20.2 483 
ACOSTA-SISON, HONORIA. Pelvimetry and Cephalometry 
among Filipinas 423 ee Ao ea ieee - 493 
- GIBSON, R. B. A Note on the Physiological Action of the | 
ProteoSes ......--00+--2 AHL Ate UMAY BLN FORT Beene lS Wi Awa 499 
GIBSON, R. B. and CONCEPCION, ISABELO. The Lympha- 
gogic Action of the Philippine Mango, Mangifera indica 
Linneus te eed a Sn ee a a ee 503 
CONCEPCION, ISABELO. Observations on Mango Rash........ .. 509 
BARBER, M. A. Milk Poisoning due to a Type of Staphylococcus 
albus Occurring in the Udder of a Healthy Cow... 515 
REVIEWS echt Sp hd ekg Rn ae ae Rar aed 
Index, Title-page, and “Contents 2 ope se cectcdeensoe he 
U. Ss. 
The ‘‘Philippine Journal of Science’”’ is issued as follows: currenoy 
Section A. Chemical and Geological Sciences and the Industries... $2.00 
Section 'B. Tropical Medicine ‘oi-s): aoe ce a ae 3.00 
Section: Ox Botany he es ee Bae a ELSON HO UT 2.00 
Section D. General Biology, Ethnology, and Anthropology (Sec- 2. 
tion D bégan with Volume Vii Oe a Bula 2.00, .-) 
Entire Journal, Volume II, III, IV, or V -.-.-..-..-.- 5.00 
Entire Journal, beginning with Volume VI * 7.00 
Single numbers of Volume [I --__--..-.....-------22.... ‘ 75 
Single numbers, (except of Volume 1)  -.-.-.22-2 220-22 cn nee e een eee 50 
Volume I, 1906 (not divided into sections). and supplement, sold 
only with a complete file of section A, B, or C.....00.2.. 220.0... 2--- ee 10.00 
Supplement to Volume 1 (Botany) 2.22.25... cccncseceseeeennteeenensenenuenee 3.50 
Volume I (without supplement), sold only with a complete file of , 
Section: As) By Or Oe es ty OS he Se a aie he Ck) 6.50 


Each section - separately paged and indexed. 


Publications sent in exchange for the Philippine Journal of Buber t 


should be addressed: Library, Bureau of Science, Manila, P. I. 


Subscriptions may be sent to the Business MANAGER, Philippine Jour- 
nal of Science, Bureau of Science, Manila,.P. 1., or to any of the agents = 


listed below: 
‘ AGENTS 


The Macmillan Company, 64—66 Fifth Avenue, New York City, U. S. Aas | 


Wm. Wesley & Son, 28 Essex Street, Strand, London, W. C., England. 
Martinus Nijhoff, Lange Voorhout 9, The Hague, Holland. 


Mayer & Miiller, Prinz Louis Ferdinandstrasse 2, Berlin, N. W., Ger- 


many. 


Kelly & Walsh, Limited, 32 Raffles Place, Singapore, Straits Settlements, 


A. M. & J. Ferguson, 19 Baillie Strect, Colombo, Ceylon. 
Thacker, Spink & Co., P. O. Box 54, Calcutta, Indin. 


Entered at the post-office at Manila, P. |., as second-class matter 


{ 


i746")
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