No. 80.— OCTOBEB, 1904

DEPARTMENT OF THE INTERIOR

BUREAU OF GOVERNMENT LABORATORIES

BIOLOGICAL LABORATORY

I. DOES LATENT OR DORMANT PLAGUE EXIST

WHERE THE DISEASE IS ENDEMIC

By Maximilian Herzog, M. D., and Charles B. Hare

SERUM LABORATORY

II. BRONCHO-PNEUMONIA OF CATTLE: ITS
ASSOCIATION WITH B. BOVISEPTICUS

By Paul G. Woolley, M. D., and Walter Sorrell, D. V. S.

III. REPORT ON PINTO (PANO BLANCO)
By Paul G. Woolley, M. D.

CHEMICAL LABORATORY

IV. NOTES ON ANALYSIS OF THE WATER FROM

THE MANILA WATER SUPPLY
By Charles L. Bliss

serum LABORATORY

V. FRAMBCESIA : ITS OCCURRENCE IN NATIVES

OF THE PHILIPPINE ISLANDS.
By Paul G. Woolley, M. D.

MANILA

BUREAU OF PUBLIC PRINTING

23394 1904

LETTERS OF TRANSMITTAL

Depaktment of the Interior,
Bureau of Government Laboratories,
Office of the Superintendent of Laboratories,

Manila, P. I., September 30, 190^.
Sir: I have the honor to transmit herewith, for publication in
one bulletin of the Bureau of Government Laboratories, the follow-
ing: I. Does Latent or Dormant Plague Exist Where the Disease
is Endemic? II. Broncho-Pneumonia of Cattle: Its Association
with B. hovisepticus. III. Pinto (Paiio Blanco). IV. Notes on
Analysis of the Water from the Manila Water Supply. V. Fram-
boesia: Its Occurrence in Natives of the Philippine Islands.
I am, very respectfully,

Paul C. Freer,
Superintendent of Government Laboratories.
Hon. Dean C. Worcester,

Secretary of the Interior, Manila, P. I.

Department of the Interior,
Bureau of Government Laboratories,
Biological Laboratory, Office of the Director,

Manila, P. I., July 15, 190J^.
Sir: I have the honor to transmit herewith and to recommend
for publication a paper entitled "Does Latent or Dormant Plague
Exist Where the- Disease is Endemic ?" by Dr. Maximilian Herzog,
Pathologist Biological Laboratory, and Mr. Chas. B. Hare, Assistant
Bacteriologist.

Very respectfully, Eichard P. Strong,

Director Biological Laboratory.
Dr. Paul C. Freer,

Superintendent Government Laboratories, Manila, P. I.

3

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Part I.

DOES LATENT OR DORMANT PLAGUE EXIST
WHERE THE DISEASE IS ENDEMIC?

By Maxtmttjak Heezog, M. D., Pathologist Biological Laboratory, and
Chas. B. Habe, Assistant Bacteriologist.

On August 21, 1903, Mr. Henry A. Blake, (1) governor of
Hongkong, addressed a communication to the secretary of state for
the colonies, entitled "Bubonic Plague in Hongkong : Memorandum
by His Excellency the Governor, on the Eesult of the Treatment
of Patients in Their own Houses and in Local Hospitals during
the Epidemic of 1903." The writer of the memorandum makes
some very startling assertions as to the danger of the spread of
plague by animals of the most varied kind, and also comes to the
amazing conclusion that there existed in Hongkong during the
period of time intervening between June 23 and July 10, 1903,
between 8,000 and 9,000 or more individuals among the native
population in which plague bacilli were present in the circulating
blood in spite of the absence of all clinical symptoms of the disease.
The governor calls this condition "latent plague" and considers it
a potent factor in the spread of the infection, and a factor which
can not of course be reached by the ordinary methods employed to
limit the spread of and possibly to suppress plague.

Fully to understand the statements of the governor of Hongkong,
it will be well to quote a few paragraphs from his memorandum,
which read as follows :

We have from Professor Simpson's report evidence that pigs, calves,
sheep, monkeys, geese, ducks, turkeys, hens, pigeons, and rats are sus-
ceptible to plague, which may be contracted by food or by inoculation direct,

5

or by means of suctorial insects. To this list the examination mentioned
above adds bugs, spiders, flies, and cockroaches. I may add that quails
kept in the market for sale were also found to be infected. In paragraph
22, page 100, Professor Simpson points out that domestic animals suffer
from chronic plague, and surmises that this is probably one of the
bridges by which the interval of the attacks in man is connected. I
have for a considerable time been of the opinion that man himself is
subject to chronic plague, which may either pass away after a con-
siderable time or continue dormant over the winter months, regaining
activity with the annual movement of spring, when the curve of the
epidernic is almost constant. This opinion was strengthened by the fact
that in August, 1899, the body of a Chinese lift man at Queen's buildings
who was accidently killed when attempting to enter the lift while in
motion was found to contain plague bacilli. A similar result followed
the examination of a man who on the 4th of March, 1901, was killed
at Tal Koo Sugar Works by a bag of sugar falling on his head from a
height of 20 feet; while on the 2d of April, 1903, in the body of the
chief steward of a ship lying in the dock, fovmd floating with a large
wound on the head, were also found plague bacilli. Early in June several
men from H. M. S. Ocean were sent to the naval hospital, suffering from
pneumonia; on examination of their blood seven were found to be
suffering from mild cases of plague. In like manner two officers of the
Sherwood Foresters who developed feverish symptoms were, on having
their blood examined, found to be similarly affected. In the "Boletin
Official" of Macao, containing the report on the plague epidemic, 1895,
Dr. Gomes de Silva, the medical officer who published the report in 1895,
stated that during the height of the epidemic he had discovered plague
bacilli in his own excreta.

(21) In June I directed Inspector Gidley to obtain as many specimens
of blood as possible, on slides secvired from the Government bacteriologist.
He obtained 110 specimens from men, women, and children taken at
random. These slides were sent to Dr. Hunter for examination, who
reported that in five slides he found plague bacilli, and in seven slides
bacilli were present in considerable numbers, some of which showed
bipolar staining. They were not sufficiently distinctive, however, to be
regarded as B. pestis. These slides were obtained between the 23d of June
and 10th of July. Since they Avere obtained there were but three cases of
plague in the district, from none of which a specimen of blood was taken.

(22) I am not unmindful of the fact that these reports were the result
of microscopic examination only. But the examination was the same as
that on which a gi-eat many of the cases treated in the Kennedy Town
Hospital were sent to that institvition where their cases ran the usual
course of plague invasion.

(23) Now, putting aside the seven doubtful slides, it will be seen
that of those people examined at random 4.54 per cent were found to be
infected with plague though to all appearances perfectly healthy. If we
exclude all the well-to-do, and take the working coolie population alone, they

probably number 180,000, and, assuming the same average amount of
infection, there are among that class alone 8,172 persons at present infected
in Hongkong. If even a quarter of that average be accepted for the
105,000 inhabitants of the superior class the number of infected will be
increased to 9,634. In Appendix G ^ will be found the nvimber of rats
examined in each month of the present year with the proportion of the
infected rats. I am afraid that the incidents mentioned in paragraph 5
weakens deduction as regards Hongkong. But, from whatever source the
rats were procured, the proportion of infection in June was 9 per cent
or 4.46 per cent more than the percentage of the slides examined, or, if
doubtful cases mentioned by Dr. Hunter be included, 1 per cent less,
while in January the proportion falls to 0.8 per cent. This being so, with
the complete circle of vermin, insects, food, rats, domestic animals, and
man all infected in possibly similar — possibly different — proportion, it
appears to me unsound to concentrate attention upon the rat as the
principal means of bridging over the dormant season.

It appears that Governor Blake, after writing the above, felt the
great danger of coming forward with so sweeping an assertion, and
in the introduction to his memorandum he himself makes an
appeal for a more through scientific investigation of the hypothesis
of the existence of latent or dormant plague among the natives of
countries where this disease is endemic. He says :

My hypothesis in paragraph 23 may not bear the light of scientific
investigation, and, as the hypothesis of a layman, may not carry much
weight, but I venture to submit that it is worthy of scientific inquiry,
for while a timely glass of water may prevent a great conflagration, and
plague at its first introduction may be stamped out by immediate segrega-
tion and thorough disinfection, its endemicity once established this is no
longer practicable, and, if the hypothesis of dormant or chronic plague
in man be ultimately proved to be correct, it is difficult to see how
quarantine for even ten days can prevent its annual recurrence, or how
any practicable examination of departing passengers can prevent its
export from the plague center and possible dissemination elsewhere if
suitable conditions for its propagation be present. What the remedy or
what the necessary precaution, I leave it for scientific men to determine,
but if my hypothesis results in a wider radius of investigation the
experiment will not have been useless.

THE EESULT OF BLOOD EXAMINATIONS IN CASES OF PLAGUE.

It is, of course, obvious to any one versed in examinations of this
nature that a diagnosis of plague can not be made by a microscopic
examination of the blood. Such an examination may possibly be

^ Omitted in this bulletin.

resorted to in urgent situations, when a rapid clinical diagnosis is
desired, but to base far-reaching deductions upon such a micro-
scopic examination is certainly not permissible. What is necessary
in order to determine beyond doubt the presence of plague bacilli
in the circulating blood is the examination of the latter by cultur-
al methods. We have undertaken a series of such examinations
in order to determine whether or not there exists such a thing as
latent or dormant plague, as suggested by the governor of Hong-
kong. Before giving the details of these experiments it will be
well to make a survey of the work that has been done with reference
to the presence of plague bacilli in the blood in undoubted cases
of this disease.

The German Plague Commission (2), in its report published in 1899,
page 265, made blood examinations in the case of 141 plague patients,
including 17 who were in the period of convalescence varying between the
seventeenth and twenty-fifth day after the disease. These examinations
were made in the following manner: A drop of blood was obtained from
the finger, with the usual asceptic precautions, and inoculated into agar
tubes, while at the same time a cover-glass preparation was also made.
It was found that in a number of cases where the culture method furnished
positive results, the mere microscopic examination failed to demonstrate
the presence of the bacilli. Of 124 patients whose blood was obtained
during the climax of the disease, in 81 even repeated examinations did not
demonstrate the presence of plague bacilli, in 10 the bacilli were encoun-
tered only once, while in 33 they were repeatedly found in the blood. Of
81 patients the examinations of whose blood were always negative, 52
recovered and 29 died. Of 10 cases, in which there was a positive result
only once, the other examinations being negative, 8 died and 2 recovered.
It is interesting to note that in one the bacilli could be detected two and
three days before death, while twelve hours before the crisis and at the
post-mortem examination it was impossible to find them. Seventeen
convalescent patients invariably failed to show any bacilli in the circulat-
ing blood.

Zoholotny (3), in his researches on plague, says that the bacilli are
found in large numbers in the blood of animals sick with the disease,
but that in the case of human beings the bacteria are much less abimdant
and sometimes can not be found at all.

Musehold (4), quoting the work of Albrecht and Gohn, reports that
the latter examined 122 cases of undoubted plague. In 55 the bacilli were
found by the culture method in the circulating blood during life. Four
of these 55 cases recovered, and in 2 of them the bacilli were present in
large numbers in the circulating blood. In the case of the 51 patients
who died the bacilli were found in considerable numbers in the blood on
the day of death as well as on the previous one.

Cairns (5) studied the blood of patients during an epidemic of plague
occurring in Glasgow in 1900. He gives the results of examinations made
inter vitam on cases which subsequently terminated fatally. Four of these
may be cited in connection with our investigation. In three fluid dravra
from the buboes developed pure cultures of Bacillus pestis, and only in one
of the four fatal cases was it possible to obtain cultures of the bacilli from
the blood during life. The other three cases gave negative results. In one
of these, seven daily consecutive examinations were made, as well as one
shortly before death, all of them proving negative.

Calvert (6) studied two epidemics of plague in Manila in 1900 and
1901. He found that the clinical examination of the blood for Bacillus
pestis gave unreliable results, it being impossible to place any weight on
the negative findings. He made his examinations by taking the blood
from the ears of the patients at intervals of four hours, and examining
it in smears as well as by the culture method. This plan was followed
until the death or recovery took place. Thirty-six cases, 4 of which
recovered, were examined in this manner, most of them being followed to
autopsy, when the plague organism was demonstrated by culture and
even by animal inoculation.

This author gives the following table of positive findings of the bacilli
in the blood:

Per cent.

24 hours before death in 31 cases , 100

48 hours before death in 7 cases 22

72 hours before death in 5 cases 16.12

96 hours before death in 2 cases 6.45

120 hours before death in 1 case 3.22

On looking over the table it appears that the plague bacilli could
be found in all fatal cases twenty-four hours before death, but that
forty-eight hours before death the percentage of positive findings
was much smaller. In searching for the bacillus in the blood of
plague patients who finally succumb to the disease, the chances of
finding the microbe five days before the fatal termination are only
one out of thirty. All of this shows that even in fatal cases the
plague bacilli are not found in the peripheral blood at an early date.

Jennings (7), in his Manual of Plague, page 112, says that it is ex-
tremely rare to find plague bacilli in the blood in large numbers except
immediately before death. Their absence, therefore, in the early stages
of an attack is frequent, and must not be regarded as a negative diagnosis of
plague.

Terni (8), who studied the plague in Rio de Janeiro, in an excellent
article on the disease makes the following statement:

"The examination of the blood is by no means reliable. It is astonish-
ing that Galeotti places any value in blood examinations in the diagnosis
of early plague."

10

Terni found that in many cases which were diagnosed as plague
septicemia, examinations of the blood, both microscopic and by culture
method, were negative. This was true even at the post-mortem examination,
because the bacilli were exclusively localized in the lymph channels.
Even in the most profound cases, in individuals particularly predisposed
to the disease, the bacilli were found in the blood only in moderate numbers.
Their presence could never be compared with what has been found to be
true in connection with other septicemic microbes, such as anthrax or
diplococci. A multiplication of the plague bacilli is found only in excep-
tional cases in the circulating blood during the agonal stage.

One of us has been studying for some time the morbid anatomy
and histo-pathology of a number of cases of plague. These studies
appear fully to confirm the statement of Terni to the effect that
plague, as a rule, is not to be looked upon as a true septicemia, but,
on the contrary, as an infection particularly confined to the lym-
phatic system. Even in cases where sections from the lymph glands
contain innumerable bacilli, the lumina of the blood vessels are
generally free from such microbes. In fact, in the study of sections
from all of the internal organs when plague bacilli are seen they
are always found in the lymph channels or lymph spaces and not
in the blood vessels.

Powell (9) has recently reported the result of 3,400 blood examinations
of febrile diseases in Bombay. Most of these cases were malaria, but 117
of them were plague. In only 15 of the latter were the bacilli easily seen
in blood smears. With reference to the finding of the bacilli in the blood
in cases of plague, the author says: "As regards the recognition of the
plague bacilli in the finger blood, for some years I was very sceptical
about the reports of certain medical men, and until within the past
eighteen months had been unable to detect the bacilli except on cultures.
At the beginning of this year there was a particular type of septicsemie
plague, in which the bacilli were found in every case. Such cases in my
experience always died. One ease seemed to be convalescent and had a
normal temperature for three days, but suddenly died. Plague bacilli
were found eleven days before death."

The Indian Plague Commission, speaking of the bacteriologic
diagnosis of plague by microscopic examination of suspected mate-
rial, makes the following statements:

(156) In the case of blood derived from a patient who is suspected of
suffering from plague, the detection of bacteria possessing the morpholog-
ical characters (of the plague bacillus will (especially if these are present
in large nmnbers and when it is determined that these become decolorized
by Gram's process) be conclusive evidence that the patient is suffering

11

from plague. The mere finding of a few isolated bacteria arranged together
as diplocoecal forms can not, especially when Gram's test has not been
applied, be accepted as establishing the diagnosis of plague. We have
in view here in particular certain suspected cases which occurred in
Calcutta in 1896, in which it was claimed by Professor Simpson that the
diagnosis of plague was confirmed by his bacteriological examinations.
We would note with regard to these first that in some of the suspected
patients only a few isolated diplocoecal forms were found after long search-
ing through a number of films prepared from the blood. Again, in view of
some of the figures reproduced in the record of Dr. Simpson's evidence,
particularly Figs. I,iVI, C, and F, it is important to note that it does
not seem to have been determined whether bacteria became decolorized when
treated by Gram's process, and, lastly the fact must not be lost sight of that
saprophytic diplococci of various kinds are widely distributed in nature,
and that there is always a possibility of microscopical preparations con-
taining as contaminations a small number of such diplocoecal forms.

In looking over these statements (all that we can find in the
literature at our disioosal), one is certainl}^ impressed with the fact
that the finding of plague bacilli in confirmed cases of the disease,
except very shortly before death or in the rarer cases of plague
septicaemia, is the exception and not the rule. Indeed, we should
be mindful of the fact that plague, as a rule, is not a septicaemia
but a bacterial infection localized in the lymphatic system. It is,
therefore, from a purely theoretical standpoint, highly improbable
that there should exist a dormant, latent clinical form of the
disease in which the patients harbor the bacilli in the circulating
blood.

Our investigations to determine whether such is the case or not
were made on a number of native Filipinos and Chinese of this
city. We attempted to get material which, if, latent plague exists
at all, would give us some evidence to this effect, therefore we
selected natives from houses in which plague cases had occurred.
We examined a number of the inmates of Bilibid Prison, par-
ticularly such as were under the most unfavorable hygienic condi-
tions, namely, insane prisoners and those of the third class who were
most crowded in their quarters. Since we could not always get
natives of this description, we selected also a number which were
not under particularly unfavorable hygienic conditions, such as
native police officers and native members of the Constabulary,

While plague has never at any time been as widespread here as
in Hongkong, a sufiicient number of cases have occurred to make

12

it clear that the disease is endemic, though fortunately not markedly
epidemic.

According to the monthly sanitary reports of the Board of
Health plague has prevailed in Manila since 1900 to the follow-
ing extent:

Prevalence of plague in Manila, 1900 to 1904.
1900.

Month.

Chinese.

Filipi-
nos.

Ameri-
cans and
other
Cauca-
sians.

Total

number

of

cases.

Total
number

of
deaths.

January

February _

March

April

May

June

July

August

September

October

November
December.

Total _

January

February ..

March

April

.May

June

July

August

September .

October

November .
December-

Total.

27
63
111
137
55
39
29
11

485

5
20
51
91
124
54
38
26
12

6

427

1

1
1

1
1

1

April

June

Julv

1

1

1

1
1

1
1

\
2

1

1
2
1
2

2

1

2

Total

5

5

10

10

13

Prevalence of plague in Manila, 1900 to 1904 — Continued.

1903.

Month.

January

February _.

March

April

May

June

July

August

September .

October

November .
December-

Total.

Chinese.

104

Filipi-
nos.

Ameri-
cans and
other
Cauca-
sians.

90

Total

number

of

cases.

Total
number

of
deaths.

1904.

January

4

6

6

12

7
8

10

7
15
15

17

7
6
14
15
16

February . . _

1

March _ .

3

8
9

April

May

Total for five months

24

39

1

64

58

SUMMARY.

Reported in—

Plague

cases

reported.

Plague
cases
fatal.

1900-

271

485
10

198
64

199

427
10

174
58

1901

1902 _ . . _ _ ____

1903

1904 (Jan. 1 to May 31)..

It appears from these statistics that plague had completely died
out during four of the months of 1902, since during this period
not a single case came under observation. However, since August,
1902, until the time of writing the present report there has not
been a month entirely free from plague, though the figures have
generally been low, the maximum during this period being reached
in April, 1903, when 52 cases of the disease were reported.

The object in giving these figures in connection with our report
is to show that plague has been sufficiently prevalent here for
several years, so that blood examinations should furnish evidence

14

of latent plague provided that such a form of the disease exists
at all.

The figures of plague morbidity for Hongkong during the same
years are as follows :

Cases.

1900 1,086

1901 1,637

1902 540

1903 1,135

BLOOD EXAMINATIONS OF 245 APPAEENTLY PIEALTHY NATIVE
FILIPINOS AND CHINESE.

The method employed to ascertain whether apparently healthy
Filipinos or Chinese of Manila have any plague bacilli in their
blood was as follows :

The bend of the elbow, generally of the left arm, was very
thoroughly cleansed first with strong alcohol and then with a strong
solution of mercury bichloride, and finally with alcohol and sterile
distilled water. A rubber bandage was then placed around the
arm above the elbow and 1 cubic centimeter of blood was drawn
from one of the veins by the aid of a sterile hypodermic syringe.
The blood so obtained was added to 50 cubic centimeters of bouillon
in a flask. The bouillon used was prepared as usual, and, when
neutral to litmus, 0.5 gram of bicarbonate of soda was added to
each 1,000 cubic centimeters of the bouillon, making it slightly,
but decidedly, alkaline. This forms a very excellent culture medium
for plague bacilli. As a control experiment in some of the cases
about twice the amount of blood was drawn from the vein, and the
2 or 3 cubic centimeters so obtained was distributed in two flasks.
One of the latter was then immediately inoculated with a very small
amount of material from a young plague culture. This was, of
course, done to see whether plague bacilli, if present, would develop
in the bouillon in the presence of a small amount of freshly drawn
blood. It may be said that the control flasks always developed a
typical plague growth, so that evidently nothing in the arrangement
of the experiment prevented development of the plague bacilli if
any were present. The culture flasks to which blood had been
added were kept either in a dark chest at room temperature or in an
incubator which was fairly constant at 35° C. The media were
inspected daily and when a growth developed it was examined in
stained preparations and by culture on agar.

15

The native Filipinos whose blood was examined included 33 labor-
ers from the Serum Institute and the morgue. The native servants
of the latter, where all the plague post-mortem examinations of
the city are made, are, of course, particularly exposed to infection,
and would be especially prone to show latent plague provided
that such a condition exists. These 32 cases were kindly examined
for us by Dr. E. H. Euediger, bacteriologist in the Serum Institute,
whose technique differed from the method generally used only in
that he employed a 5-per-cent carbolic acid solution to sterilize the
elbow. Dr. Euediger also examined all of his 32 flasks by stained
cover-slip preparations and by culture methods whether they
showed any change in appearance or not.

The blood examinations in all of the 245 cases were made between
March 4 and May 20, 1904 — i. e., during a period when from
thirty-five to forty cases of plague were reported in Manila.

The following is a summary of the examinations :

On March 4 there were examined 5 native Filipinos from a house in
Santa Cruz in which an ambulatory case of plague terminating in
embolism of the pulmonary artery had occurred. Specimens were taken
from 10 native Filipino police ofl&cers on April 6 and 7. Nine of these
men lived in infected districts — i. e., those in which cases of plague had
recently occurred; 1 came from a noninfected district. Ninety native
Filipinos, members of the Philippines Constabulary, were examined between
April 13 and 30. These men live in barracks, but are often free to visit
their families and friends. During the first week in May 32 native
Filipinos, laborers at the Serum Institute and the morgue, were the
next subjects investigated. In the former place the various vaccines
and sera, including plague vaccine and serum, are prepared, and in the
latter are performed all the necropsies on plague cases. Fifty-eight native
Filipinos, prisoners in Bilibid Prison, were taken on May 12 and 13. In
this place about 4,500 men are confined. Last year a number of the
inmates died from pneumonic plague, but during this one no case has
occurred there, although one of the prisoners died of pulmonary plague
four days after his discharge. Of these 58 men 16 were insane prisoners,
all in advanced stages of degenerative mental disease, and the remainder
were the so-called third-class prisoners, who lived under the most unfavor-
able conditions to be found in the prison.

On May 16 to 20 there were examined 50 Chinese, small shopkeepers,
clerks, and coolies, either from houses in which plague cases (in' one
instance two such cases) had occurred or from those adjoining.

RESULTS OF THE EXAJVIINATION.

Most of the flasks to which 1 cubic centimeter of blood had been
added remained sterile; although a few developed growths which.

16

however, were clearly contaminations from the air, such as common
molds or similar forms of life. One of the cases taken by Dr.
Euediger developed Staphylococcus pyogenes aureus; and those of
two other natives developed a bacillus which, when examined in a
stained cover-glass preparation, might possibly be mistaken for
Bacillus pestis. One of these organisms, however, in culture looked
very different from the bacillus of plague and also retained Oram's
stain. This bacillus developed in a flask to which had been added
blood from one of the insane prisoners. The other growth occurred
in a flask containing blood from a member of the Constabulary.
This organism also greatly resembled morphologically the plague
bacillus, but it kept Gram's stain, and when rubbed in large amounts
on the shaved abdomen of a guinea pig failed to produce disease.
In short, not in a single instance out of 245 examinations of persons
of whom a large percentage had certainly been greatly exposed to
plague infection did we find any evidence of the existence of plague
bacilli in the blood.

CONCLUDING EEMAEKS.

From our investigations conducted on 245 native Filipinos and
Chinese it may be safely concluded that a condition of latent or
dormant plague does not exist in Manila, and there is hardly any
reasonable doubt that it does not exist in Hongkong. There
certainly has not been furnished the slightest proof of such a
character as to stand the searchlight of exact methods of bacterial
investigation to indicate that there is such a thing as latent
human plague, with the presence of plague bacilli in the circulating
blood, in the absence of clinical symptoms of the disease.

The governor of Hongkong himself, in his memorandum, clearly
sets forth some of the circumstances which unite to make it
practically impossible to completely eradicate the disease in Victoria
City.

In Hongkong (1) (memorandum, par. 3) it is the custom of
the Chinese — if they can possibly do so — to dump human corpses
dead from plague into the street, in order that they may not be
found in the houses and thereby subject the inmates to quarantine,
disinfection, etc. In spite of measures to prevent this procedure,
the number of corpses dead from the plague and so disposed of has,
during the ten years preceding 1898, increased from 25.1 per
cent to 32,7 per cent.

17

The Chinese in Hongkong, according to the governor's memoran-
dum, offer passive resistance to the catching of rats in their houses,
being afraid that plague bacilli might be found in the rodents, as
this would lead to measures of disinfection or to the repair of their
houses. The Chinese rat catchers are said to be dishonest; they
fail properly to label the rats, so that infected houses escape detec-
tion, and they import rats from the outside of Hongkong and
label them at random. In general they are very unreliable in
their work and are actuated solely by the desire to secure from
anywhere the largest number of rats in order to obtain the premium
offered for each.

To those who know how Chinese houses are constructed [says the memo-
randum, in par. 6], it will be apparent that effective fumigation is
practically unattainable. While, even if the spraying process, scrubbing
and disinfection of clothing reached externally everything in the room,
it would not kill vermin lying deep in the joints and cracks of the tables,
chairs, and settees, or beds. Nor would it reach the vermin with which the
heads of the poorer classes of coolies are infested. But apart from this,
what took place in many cases when a case of plague was detected was
that before the constable could arrive to take charge of the house, goods
liable to injury by disinfection were removed by the door, or, if too late for
this, were taken on to the roof, always easily accessible, and deposited in
some neighboring house.

In W. J. Simpson's report (11) "On the Causes and Continuance
of the Plague in Hongkong, etc.," which is quoted in the memo-
randum of the governor, we find the following statements as to the
sanitary conditions in the Chinese quarters in the city of Victoria.

When a case of plague has once occurred in a house, there is a great
tendency in subsequent years for the same house, or that adjoining, or
that on the opposite side, or that close by, to be attacked with plague.
When plotted out on a map, the distribution of plague appears to be
closely connected with previous infection of the house or of a defined
locality, the infection having been retained in an unrecognized form in
the interval. The houses which suffer principally are, speaking generally,
the most insanitary and the oldest. It has already been mentioned,
how closely packed the buildings are in the older portions of the town,
narrow streets and high houses being the leading features, by which the
admission of sunlight and fresh air is considerably obstructed. Narrow
streets and high houses, however, are not peculiar to Hongkong; they
are to be found in other towns, with their injurious effects on health,
but in Hongkong there is moreover in the Chinese quarters a defect
in the construction of the houses which intensifies the obstruction of light.
The rooms are long and narrow, with a window at each end, the front

23394 2

18

window looking into a wide and covered veranda, and the back window
into a small open space at the back, which forms a sort of wall between
the two houses. The lower floors of many of the houses are remarkable
for their darkness, and this in a region not far from the Tropics; they are
also frequently damp.

Since the epidemic of 1894 many of the lower floors of the worst kind
have been changed into storerooms to contain the goods and merchandise
for which Hongkong is in entrepot. Tliese storerooms as a rule are
infested with rats, which at times find their way up to the rooms on the
higher floors. The basements are generally rat ridden, both floors and
walls, and from the walls being often hollow it is easy for rats to reach
the upper floors.

The admission of sunlight into the dwelling rooms of Chinese tenement
houses is still further obstrvicted by the subdivision into several cabins
or compartments, sometimes nmubering up to six, which every room is
subjected to. Each cabin is let out to a separate tenant and not infre-
quently accommodate a separate family. These compartments or cubicles
are windowless rooms, and are often so dark that it is impossible for any
one, coming directly from the light outside and drawing or opening the
door of the cubicle, to see at once whether it is occupied * * *. Some
attempts have been made to improve this state of things by limiting the
height of the subdividing Avails to six feet. The condition Avhich obtained
before this imj^roA^ement has made it somewhat difficult to realize, for
what I am describing is that which now exists. Fresh air and sunlight
never get into the cubicles except perhaps the compartment at each end
of the room opposite the window. The subdivision of a single room
into a number of rooms called cubicles is an ingenious device for crowding
together a large number of people into a small space and securing a
correspondingly large rental, but it is an arrangement Avhich engenders
disease and favors its spread. There is no doubt whatever that every such
windowless cubicle is unfit for human habitation and should not be
permitted * * *. Probably another cause for the continuance of the
plague, besides the insanitary condition of the houses referred to, is the
very inadequate number of latrines and urinals with which Hongkong is
provided. The number of public latrines appears to be twenty-nine belong-
ing to the Government, and seventeen to private OAvners. The total
number of seats is 1,202. Most of them have urinals attached, and in
addition there are three small public urinals in the town. Seeing that all
the men and boys go to the public latrines, there are no sanitary appliances
in the houses except earthen pots, AA^hich are used exclusively by the
women and children. The total inadequacy of the latrine accommodation
provided is obvious. It is not one seat to one hundred of the male popula-
tion. On the KoAvloon side of the colony the" latrine and urinal accommoda-
tion is still more deficient. Large blocks of houses have been built, and
not a single latrine or urinal provided by the builder of the block. It is
impossible under these conditions that the ground should escape being
seAvage polluted * * *. The existing latrines are far from being models

19

of what they should be. Tliey are in fact insanitary in structure and
deficient in light and ventilation.

Quite recently Surgeon-General Evatt, P. M. Q.^ His Majesty's
troops, Hongkong, has been quoted by the daily papers in an inteir
view in which he speaks in the strongest terms of the insanitary
conditions, which he holds responsible for the continued prevalence
of plague in Hongkong. He calls Victoria City the plague-
distributing center of the world, a standing menace to the human
race. So we have the most convincing evidence, both official and
unofficial, as to the vicious sanitary conditions in Hongkong. These
furnish the soil in which plague thrives, from which it can not be
completel}^ eradicated, and from which it breaks forth again and
again in menacing epidemic form.

From our own investigation carried on in Manila, we have
certainly good reasons to deny the existence of latent or dormant
plague in our city. The statements of those who have looked into
the conditions in Honkgong likewise appear unfavorable to the
theory of the governor, and they offer an explanation more in accord
with what we do know positively as to the nature of plague infec-
tion, and as to the spread of infectious diseases in general.

REFERENCES TO LITERATURE.

(1) Henry A. Blake. Bubonic Plague in Hongkong; Memorandum
by His Excellency the Governor on the result of the treatment of
patients in their own houses and in local hospitals during the epi-
demic of 1903. Hongkong. Printed by Noronha & Co., Govern-
ment printers, 1903.

<2) Bericht der Deutschen Pest Commission. Gaffky, Pfeiffer, Sticker, and
Dieudonne. Arbeiten aiis dem Kaiserlichen Gesundheitsamte. Vol.
16, 1899, p. 265.

(3) Zobolotny. Recherches sur la Peste. Archives des Sciences Biolo-

giques. St. Petersburg, 1901, vol. 8, p. 81.

(4) MusEHOLD. Die Pest. Berlin, 1901, p. 150.

(5) Cairn. On the Agglutinating Property of Blood Serum in Cases of

Plague. London, Lancet, 1901, June 22, p. 1746.

(6) Calvert. Plague Bacilli in the Blood. Centralblatt fuer Bakteriologie.

I Abth. Originale, vol. 33, No. 4, p. 247.

(7) Jennings. Manual of Plague. London, 1903, p. 112.

(8) Terni. Studien ueber die Pest. Zeitschrift fuer Hygiene und Infek-

tionskrankheiten. 1903, vol. 44, p. 151.

(9) Powell. The Blood Examination in 3,400 Cases of Febrile Diseases in

Bombay. Indian Medical Gazette. Calcutta, Feb. 1904, vol. 39, No.
2, p. 41.

20

(10) Indian Plague Commission. Report, Vol. V, p. 60. London, 1901.

(11) Hongkong: Report of the Principal Medical Officer for the Years

1900, 1901, 1902, 1903. Hongkong. Printed by Noronha & Co.,
Government printers.

(12) Simpson. Reports on the Causes and Continuance of Plague in Hong-

kong and Suggestions as to Remedial Measures. London, Waterlow
& Sons, Limited, 1903.

Part II.

BRONCHO-PNEUMONIA OF CATTLE: ITS ASSO-
CIATION WITH B. BOVISEPTICUS.

By Paul G. Woolley, M. D., Director of the Serum Laboratory, and.
Walter Sorbell, D. V. S., Y eterinarian Serum Laboratory.

The reason we care to dwell upon these cases is not that the
above-mentioned pulmonary affection is uncommon or not well
known, but because of its relation, here at least, to hemorrhagic
septicaemia.

It is agreed by all authors that the causes of catarrhal pneumonia,
in animals as well as in man, are not always the same. However,
in general the chief ones are also those of acute bronchitis. Follow-
ing this it happens, either because of swelling of the mucous
membrane, or by aspiration of bronchial secretion, that the small
subdivisions of the bronchi became plugged, and a condition of
atelectasis arises in that part of the lung supplied by the affected
bronchioles. Then because of the locus minoris resistentice fur-
nished by the atelectatic part, and because of the usual presence of
infectious material in the bronchial secretions and in the aspirated
material, inflammatory processes occur, which may be limited or
which may spread by continuity to the neighboring tissue. The
changes following atelectasis may either be a gradual atrophy of
the affected parts of the lung, or following infection, bron-
chiectasis, abscess formation, pleurisy, gangrene, etc., resolution,
or calcification. The disease occurs especially in young animals and
in older ones whose physical condition is below par. In young
animals it may be epizootic.

The symptoms are said to vary. Schneidemiihl says that the

21

22

first signs point to acute bronchitis, upon which ensue pulmonary
symptoms, such as increased respiratory frequence and cough.

The course of the disease according to Diekerhoff and Schneide-
miihl is neither typical nor regular. Sometimes it progresses very
rapidly. In other cases it may last for weeks or months.

In the more chronic ones a purulent pneumonia with pleuritis
and pulmonary gangrene may ensue and death follow. It is in
this last group,, that of chronic cases, that ours belong.

These occurred in the herd of the Government Serum Laboratory,
The calves were imported for use in the preparation of vaccine
virus. The cattle were used in the preparation of antirinderpest
serum. All of the calves originally came from China, and upon
arriving in Manila had been treated with prophylactic doses of
antirinderpest serum and kept under careful observation for several
weeks before they were used for the intended purpose. At the time
of vaccination there was no indication of disease in any member
of the herd, their appetites were good, they were in excellent
physical condition, and their temperature curves were normal.
(Inasmuch as tuberculosis has never been observed in cattle here
the vaccine calves are not slaughtered after collecting the virus, but
are kept on hand until disposition can be made of them in some
other way. As a rule, the physical condition of the animals
improves after taking the vaccine. When this is not the case
slaughter is resorted to when there is no particular reason, as in
the present instances, of allowing the animals to live. )

The steer mentioned below was also imported from China for
use in the preparation of antirinderpest serum, but its condition
was never satisfactory for this purpose.

The histories follow :

Case I.

Calf No. JjSl Avas received at the laboratory on January 12, 1904, and
received 100 cubic centimeters of antirinderpest serum at once. It re-
mained apparently well with a normal temperature until January 24,
when the latter rose to 40.7° C. The next day the temperature was
40.8° in the afternoon, and the following day it reached 41.2°. From
this point it gradually declined to normal on January 30. On February
1, 5 cubic centimeters of virulent rinderpest blood was given to the animal,
and on the third day after, it was vaccinated. Following this operation,
there was what at first appeared to be the usual temperature of vaccinia,
which reached 40.6° on the fifth day after vaccination. But this tempera-
ture instead of declining persisted with slight variations between 39° C.

23

and 41° C. until March 1, when it became normal and stayed so until death,
March 24, 1904. On several occasions the blood of the animal was examined
for trypanosomas with negative results.

During the course of the disease, the animal gradually lost weight in
spite of a reasonably good appetite, and as it progressed the appetite
became impaired and the coat rough and stairy. There was no cough,
nor in fact any other notable symptoms than the gradual wasting.

The autopsy was made about twelve hours after death. The only
appreciable lesions present were in the lungs. The other organs were in an
apparently healthy condition, with the exception of the preseapular
lymph glands, which were enlarged and edematous.

The apices of the lungs were chiefly affected. These portions of the
organ were almost completely solidified. The surface of the hepatized
portions were mottled with yellowish-white areas which stood out dis-
tinctly upon a reddish-purple ground. In palpating, the finger was able
to detect that these lighter areas were firmer than the darker colored
portions of the lungs, which had an edematous consistency. Over several
of the nodules there was a thin, fibrinous membrane under which the
pleura was congested and roughened. Upon section, the affected portions
of the organs cut with increased resistance, and from the incisions a
frothy serum oozed. The general color of the surface was dark red, mot-
tled by the grayish yellow of the sometimes almost caseous nodules.
Sections of the latter had all the macroscopic characteristics of the gTay
hepatized tissue of pneumonia, being gi'anular and dry. In an occasional
one the process had gone beyond the simple hepatization and the center
had softened, producing a rather creamy material.

From several of such tubercles, cultures were made upon agar and in
bouillon, and pieces of the tissue were placed in Zenker's solution, and
absolute alcohol, for further study. Smears made from the pulmonary
nodules showed a few small ovoid bacilli, which stained with the ordinary
anilin dyes and were decolorized both by Gram's and Gabbett's methods.

Cultures of organisms were obtained which had all the characteristics
of the bacillus described by one of us as the cause of an epidemic of
hemorrhagic septicaemia among the carabaos of the Government corrals
during the past year. The chief characteristics of this organism were its
polar staining, rounded ends, nonmotility, and occasional encapsulation. It
grew invisibly on potato, did not produce gas in either solid or liquid
glucose media, did not coagulate or acidify milk, did not form spores or
liquefy gelatin, but did reduce nitrates and also gave the Indol reaction.
It was pathogenic for guinea pigs, causing death within twenty-four hours
after intraperitoneal injection of 1 ciibic centimeter. It was the Bacillus
bovisepticus of Kruse.

The tissues preserved for sectioning were embedded in celloidin, and
sections from these were stained with haematoxylin and eosin, by Gram's
method and by the carbol-fuchsin-acid one for tubercle bacilli. The stained
sections showed a generally edematous condition of the parenchyma. The
trabeculse were somewhat thickened and the fibrous tissue of the affected
portion of the lungs was generally increased. The epithelial linings of the

24

bronchi were convoluted and hyperplastic, the cells being in many places
three or four layers thick, and in the dilated lumina there were considerable
accumulations of epithelial cells, polymorphonuclear leucocytes and a
minimal amount of fibrin.

The air spaces of the affected lobes which were not involved in the con-
solidation were filled with a granular material with which there were
occasional desquamated cells and leucocytes. The peri-bronchial tissue was
the seat of well marked round cell infiltrations and the peri-bronchial
connective tissue was considerably increased.

Sections of the small nodules of consolidation showed that the chief
change in these was a coagulation necrosis. The centers of these areas
were crowded with leucocytes and granular, cellular detritus enmeshed
in fibrin. The walls of the air cells were perceptible as faintly pink-
stained bands in which no nuclei or only shadows could be seen. As the
periphery of these areas was approached, the lines became more distinct,
and about the latter was a zone of congestion. There were no giant cells
seen in any sections, and no "acidfast" bacilli could be found. There were,
however, a number of very short bacilli between the cellular contents of the
abscesses.

Ca.SE II.

Calf No. Ji23 was received from Hongkong on January 6, 1904, and
given a preliminary prophylactic dose of 100 cubic centimeters of anti-
rinderpest serum. It was vaccinated on January 13, after which its
temperature rose to 40° C. where it remained with slight remissions
until January 19, reaching normal on the 20th. On January 21 it re-
ceived 10 cubic centimeters of virulent rinderpest blood, and following
this the temperature again rose and varied between 39.2° and 40.6° for
the next eight days. On February 1 it received 50 cubic centimeters of
virulent blood, and following this there was again a rise, which, however,
was transient. From this time on the temperature remained within normal
limits. Death occurred April 30, somewhat more than three months after
the arrival of the animal at the laboratory.

During the last month or five weeks of its life a steady and gradual
decline was evident. The animal lost weight in spite of a constantly fair
appetite. Upon several occasions the blood was examined for trypanosomas,
each time in vain. No cause could be found for the wasting. There was
no cough, in fact no other symptoms than the gradual emaciation, and
increasing weakness.

The day before death the animal was unable to stand but was lying in
its stall in the hospital shed, eating the grass before it. It died the
following night and was found in the morning in its sleeping position.

Upon opening the thorax a considerable amount of clear serofibrinous
fluid gushed out. The peritoneal cavity contained no fluid and was
apparently normal. The subpleural and mediastinal tissues were edematous
and gelatinous in appearance. The prescapular glands and periglandular
tissues were also edematous. The liver appeared pale but otherwise nor-
mal. There were a few petechise in the capsule of the spleen, which was of
normal size and consistence.

25

The cecum contained a number of oesophagostomas and fasciolas.^ There
were no gastro-intestinal hemorrhages or ulcerations.

The lungs were the seat of a patchy hepatization, which involved about
one-half of the entire pulmonary tissue, chiefly the apices and anterior
lobes. In the hepatized areas, and perceptible to the eye and finger, upon
the surface of the lungs, were small nodules. These were small and
paler than the adjacent tissue, which had an almost purple color. Section
of these organs showed that the fibrous tissue was generally increased in
the affected portions, the trabeculse being quite prominent. The surfaces
of cut sections showed mottled gray and red, the gray being most apparent
in the centers of the lobules. Certain of these gray areas were quite dry
and almost caseous, and were as large as a navy bean.

From these nodules cultures of a nonmotile polar-staining bacillus
which did not stain by Gram's method and did not form spores were obtained
on agar. The colonies in agar varied in size from 0.5 to 1 millimeter in
diameter, were thick and opaque at their centers, were not chromogenic,
had smooth, regular edges and were somewhat sticl<y and mucilaginous.
On agar slants after twenty-four hours the growth was colonial, but with a
tendency to confluence. The water of condensation Avas clouded.

On glycerin-agar the growth was more luxuriant, quite thin but almost
opaque, and so moist that it was inclined to run down the surface of the
slant.

Milk became just perceptibly acid after seventy- two hours, but without
coagulation or reduction of the litmus. The growth on potato was invisi-
ble. In glucose broth and agar no gas was formed. In Dunham's peptone,
to a liter of which 5 cubic centimeters of a .5-per cent solution of potas-
sium nitrate had been added, a brillant cholera red reaction could be
obtained after twenty-four hours. ^ After seventy-two hours a pellicle
appeared in the ordinary bouillon; the medium was diffusely clouded and
a flocculo-gelatinous sediment was thro^vn down.

Of a twenty-four-hour-old broth culture, 1 cubic centimeter was injected
into the peritoneum of a healthy guinea pig. The animal survived this
treatment.

Cellodin sections cut from material fixed in Zenker's solution, and
stained with hematoxylin and eosin, showed changes that correspond in all
general respects with those described in the previous case.

Case III.

Calf No. 415 was received at the laboratory on December 29, 1903, and
given the usual prophylactic dose of 100 cubic centimeters of antirinderpest
serum. The temperature remained normal until after vaccination, which

^ Specimens of these parasites were sent to Chas. Wardell Stiles for
identification.

^ This method was suggested to us by Dr. W. B. Wherry. For some time
we have all been troubled by the inconsistency of the peptone used in
making Dunham's solution. Dr. Wherry, however, found that a constant
reaction could be obtained by adding traces of nitrates to that fluid.

26

was done eight days later. Following this, on the third day, the tempera-
ture rose abruptly to 41.6° C. and then gradually fell to normal within
the next week. On January 20, 1904, it was given a subcutaneous in-
jection of 10 cubic centimeters of virulent rinderpest blood, following
which there was a reaction of 1.4° C, after which the temperature fell
to normal and continued so to the time of death on May 14, 1904, almost
five months after the animal had been received. At no time were trypa-
nosomas found in the blood. The clinical history in the case was not unlike
that in the previous one; the salient points being gradual loss of weight in
spite of retention of appetite, normal temperature, no cough, roughened and
stairy coat, and loss of strength.

At autopsy the chief lesions, and in fact the only macroscopic ones, were
in the lungs. There was not as much edema as in the previous case, and
there was a less general pulmonary involvement. But here, as in the
other ones, the apices and anterior margin of the lungs were partially
solidified, mottled Avith red and gray, and filled with small, firm nodules,
some miliary and some the size of a large bean. Some of these on section
proved to contain a semifiuid purulent material, while others were dry
and gray.

Cultures were made as before from these nodular lesions and in this
case two organisms were isolated, one B. pyocyaneus, the other a polar
staining bacillus agreeing in general with the organism previously described,
but varying from it in some cultural characteristics.

Morphologically, it was identical with the organisms from the previous
cases. Culturally, variations were most marked in broth and on agar. On
the latter, the growth was generally composed of colonies, but these were
somewhat larger than the ones described for B. Itovisefticus. The growth
was much more luxuriant and whiter and there was a decided tendency to
confiuence on the part of the colonies.

In bouillon there was at first a diff'use cloudiness with no sediment and
no pellicle. Later, a gelatinous sediment was deposited, a very delicate
pellicle was developed, and the body of the medium became clear. There
was also a deposition of floccules on the sides of the tubes.

The cholera-red reaction was obtained after twenty-four hours. Milk
was unchanged, and there was an invisible groAvth on potato.

Intraperitoneal injection of 1 cubic centimeter of this organism killed a
healthy guinea pig in less than twenty-four hours. At autopsy an acute
hemorrhagic peritonitis was discovered, and the organism recovered.

Case IV.

Calf No. Jf64 was received at the laboratory on January 12, 1904, and
was injected with 100 cubic centimeters of antirinderpest serum. The
following day its tempertaure was 40.1°, and on the succeeding it was
normal and remained so for the next ten days. On January 20 it was
vaccinated with vacine virus, and following this its temperature rose,
reaching a maximum, 41.1°, on the day after the vaccine was collected.
Two days later it was again normal. On February 1, 1904, 10 cubic
centimeters of virulent rinderpest blood was injected subcutaneously, and

27

following this the animars temperature became quite irregular, reaching
40.5°, and with daily remissions of 1 to 2 degrees; but after twelve
days the maximum had fallen to about 39.2° C. and with smaller
remissions. This continued until death occurred. May 9, 1904, about
four months after its arrival in Manila. Up to this time it was able to
eat, but it gradually became emaciated and very weak.

At autopsy the lungs were found to be affected in a manner similar to
the condition of the others described above. In this instance, however,
the apices were alone affected over an area in each of about the size of the
palm of a hand. These were purplish-red, edematous, and contained small
areas of solidification, which appeared gi-ayish or gray in sections.
There was a fibrinous exudate over the pleural surface of some. The other
organs showed no change other than mild parenchymatous degeneration.
The body was anemic. ^

Cultures were made from the nodules in the lungs. On the original
plate cultures two types of colonies were noted: One very small, similar
to the typical colonies of B. hovisepticus, the other larger and with more
tendency to spread, and upon slant cultures to coalesce. Both of these
were studied.

The only cultural differences in these two strains were noted in agar,
upon which one showed a greater inclination to spread and coalesce, and
in broth in which one ( 464 ^ ) caused a uniform clouding with a gelatinous
sediment, while the other ( 464 ^ ) clouded the media less diffusely and
formed flocculi, which were present not only in the body of the fluid but
also on the sides and bottom of the tube. Both gave a brilliant indol
reaction. Both were pathogenic for guinea pigs.

Sections for material fixed in Zenker's solution and imbedded in cel-
loidin showed the same histologic picture as that described in above
cases.

Case V.

Calf No. Jf91 was received at the Serum Laboratory on February 24,
1904, and was given a prophylactic dose of 100 cubic centimeters of anti-
rinderpest serum. A transient rise of temperature to 41.2° C. and a
gradual return, during seven days, to normal followed this. On March 5,
1904, 5 cubic centimeters of virulent rinderpest blood was injected
subcutaneously, upon which a scarcely perceptible rise of temperature
lasting but twenty-four hours was noted. Four days later the animal was
vaccinated and again the temperature rose, remained between 39° and
40° C. during the inoculation disease, and then gradually fell to normal.
On March 25, 1904, a second injection, this time 50 cubic centimeters, of
virulent blood was made, and this gave rise to a slight increase in
temperature, which lasted but forty-eight hours.

During the following month the temperature remained above normal,

^ In the small intestine there were some round worms, which were
preserved and sent to Chas. Wardell Stiles for identification. In his
report he says that they are in all probability a new species of Hsemonchus.

28

varying from 39° to 40° C, and only dropped immediately before death.
The animal died on May 5, 1904, a little over two months after its arrival
at the laboratory.

At autopsy, done about eighteen hours after death, the upper halves
and the anterior lobes of the lungs were found to be of mottled purplish-red
color, and almost completely consolidated. In the areas of consolidation
were a few nodules, generally of a pyramidal shape with their bases
in the pleura.

The pleural surfaces of these were covered with a thin fibrino-purulent
exudate, which upon removal showed the yellowish color of the nodule. On
section the nodules appeared to be abscesses, surrounded by consolidated
pulmonary tissue, and containing a thick yellowish pus.

Throughout the consolidated portions of the lungs there were smaller,
almost miliary, firm areas. In general these were in the centers of
lobules about which the trabaculse surrounding them were increased in
size and quite prominent.

The other organs showed no marked changes.

Cultures from the abscesses upon glycerin-agar and in broth showed no
growth.

Histological examination of sections showed approximately the same
arrangement in and about the nodules as in the previous cases.

Case VI.

Calf No. Jti-5S was received at the Serum Laboratory on January 12,
1904, and given 100 cubic centimeters of antirinderpest serum. The
next day its temperature was 40.2° C, after which it fell to normal,
remained so for three days, and then gradually rose to 40.8°, afterwards
again gradually falling, the course covering a period of one week. From
this time until death the temperature remained normal, except for a
transient rise after vaccination and one following injection of 50 cubic
centimeters of virulent rinderpest blood. It died on April 27, 1904,
about three and one-half months after coming to the laboratory.

At autopsy the pulmonary lesions were similar to those found in No.
491, though less extensive, being almost limited to the apices and the
extreme exterior borders.

Plate cultures from the nodules after twenty-four hours at 37° showed
three slightly variant types of colonies. Transplants were made from each
type and studied as B. 453,^ 453,^ and 453.^

After carefully comparing their appearances in and on various media,
no other difference could be seen than that of size. B. 453 ^ was a trifle
larger, generally, than the other two. All produced typical colonies —
i. e., the ones which were larger, more opaque, and which showed a
slight tendency to coalescence.

These various differences between these strains will be discussed later.
Suffice it to say now, that, in general the organisms in their cultural and
morphologic characteristics were identical with B. hovisepticus.

Histologic examination showed a condition that agreed with that seen
in previous cases.

29

Case VII.

Steer No. 383 was received at the Serum Laboratory on January 6,
1904, and was given 100 cubic centimeters of antirinderpest serum. Two
days later it showed some indisposition, and upon examination the
temperature was found to be normal, the pulse somewhat accelerated,
respirations rapid and shallow, coat stairing, appetite diminished, and
rumination performed with indifference. Later the temperature became
irregular with an evening rise which often reached 40° C. About ten
days later, after the first examination, the animal developed a shallow,
painful cough, which became more marked as the disease progressed.
Tuberculosis, although up to that time unknown in these Islands, was
suspected and the steer Avas killed sixty-one days after coming to the
laboratory.

At autopsy a very widespread pathologic condition of the lungs was
found affecting almost the entire extent of both organs. This consisted
in a chronic bronchitis with bronchiectasis upon which a broncho-pneumonia
had been engrafted. The bronchial walls were injected and covered with
a sticky mucopurulent material. There were several bronehiectatic cavi-
ties, one the size of a goose egg, the others smaller, which had smooth
fibroid walls within which were greenish-yellow muco-caseous masses of
very foul-smelling material. The parenchyma of the lungs was studded
with small areas of consolidation, some of which had undergone softening.
The lungs were generally edematous. There were no marked changes
elsewhere save congestion and cloudy swelling.

Cultures made from the small nodules at the time of autopsy showed
no growths.

Tissues were preserved in Zenker's solution and embedded in colloidin.
Sections from these materials were stained with hematoxylin and eosin,
thionin, genetian violet ( Gram and Weigert methods ) , and by the ones
used to demonstrate tubercle bacilli.

The lungs on section showed an intense peri-bronchial infiltration, which
was composed for the most part of cells of an epithelioid character with
abundant protoplasm and vescicular nuclei. There was an increase in
the connective tissue of these accumulations, which were also more or less
regularly enclosed by fibrous capsules, making it appear as though the
epithelioid accumulations had occurred betAveen the mucous nlembranes
and the surrounding muscular layers. Outside of the encapsulating layers
were occasional lymphoid accumulations.

The mucous membrane of the bronchi was hypertrophied and cor-
rugated— the cells being tAVO or three layers thick and many of them
vacuolated. The lumen Avas filled with mucous material in AA'hich numerous
leucocytes AA^ere embedded.

In several places the mucous membrane on one side Avas hypertrophic,
while on the other it Avas flattened and atrophied. Projecting into the
lumen from the hypertrophied side Avere masses composed of small round
cells and stroma, about whose internal edges were a row of large cells
of peculiar appearance. One of these was an enormous cell whose pro-
toplasm Avas very large in amount and whose nucleus was about 3 to 4

30

times the size of a poly-morplionuclear leucyte, and vescicular. The other
ones were not as large, but were of the same type, except one large giant
cell from whose position, and the arrangement, it is plain that it is
formed by the coalescence of epithelioid (endothelial) cells. The remainder
of the lumen was filled with cells similar to those described.

That the structures of the growth were granulomata might be seen
in this same section, for the mixture of round epitheloid cells projecting
into one large bronchus was well supplied with mature and immature blood
vessels. There was no giant cell formation in these epithelioid or round-
celled nodules. The only place, apparently, where this was found was
on the lumen side of the projecting growths, and even here the occurrence
was an exception.

The trabeculse of the Imigs were increased in volume. The perivascular
connective tissue was increased and the parenchyma was injected.

It certainly seemed that in some places there was a tendency to syncytial
formation in the mucous membranes of the bronchi, especially in those in
which the peripheral groAvth had encroached largely iijion the lumens.

In other sections the profuse fibroid overgrowth was most well marked.
In such bronchi, so nearly obliterated that nothing but remnants of
mucous membranes were to be seen embedded, masses of round and
vescicular cells occurred. Giant cells could be seen in these, bvit here,
too, these were nearer the peripheral margin of the nodules, and they
seemed in a certain degree to have some relation to the smaller bronchioles
upon which the cellular accumulations had encroached.

In other sections nodules could be seen whose centers evidenced com-
mencing rhexis.

The liver showed a remarkable degree of fibroid change, evidently
originating about the bile ducts. The tissue, besides its formative elements,
containing many cells with vescicular nuclei. New gall-duct formations
Avere also noticed.

No organisms were seen in the sections that resembled glanders bacilli,
and there were no "acidfast" organisms discovered.

Cases VIII and IX.

Calves Nos. J/SG and Ji-OS. — Neither in their clinical symptoms nor in
their general condition did these animals show any marked symptoms of
disease. Their temperature kept within normal limits except after virulent
blood injections and after vaccinations. They had no cough. They ate up
to the day of death.

In the case of No. 468, which became much weaker than did No. 405,
appetite was present and the animal ate its fodder eight days after it had
been unable to rise to its feet.

The lesions were comparable to those described in the other calves, but
in both eases they were limited to the apices of the lungs. From the
lesions of each animal an organism was obtained in cultures which was
similar to the ones already described — viz, to B. hovisepticus.

The histologic examination showed approximately similar changes,
although the processes were less advanced.

31

That these cases arose in the way Diekerhofi; and Schneidemiihl
describe, there can be but little doubt. In certain animals, whose
physical condition warranted slaughtering, we have noticed small
areas of atelectiasis, which might or might not have been con-
genital, a point which we have been unable to determine. In
certain others we have found both acute and subacute bronchial
changes, with no macroscope parenchjanatous changes. Whenever
two such conditions coincided in the same animal the ideal oppor-
tunity would be afforded for the production of more serious
pathologic complex, and we imagine that this is exactly what has
happened in certain of these cattle.

It it an interesting feature of these cases that the bacilli of
hemorrhagic septicaemia should be associated with so large a pro-
portion.

Just what the duration of the disease has been in these animals
can not be determined accurately. In the steer it was undoubtedly
at least fifty days, and in the others it was probably thirty to
ninety.

Infection has undoubtedly taken place in each case following
the primary bronchitis, still later incipient pneumonic changes
setting in.

Since the organisms present in the lesions proved to be the
bacillus of hemorrhagic septicfemia, we may conclude that this
is as common an inhabitant of the respiratory tract of animals
here as it is in the United States.

Finally it may be said that from the above-mentioned facts that
these cases are not examples of chronic infection with B. dovisepti-
cus, but simply of an implanted infection in the course of other
pathologic conditions, or perhaps in certain cases of a terminal
infection. It seems probable, from our experience, that when
primary infection with this organism occurs the disease runs a
somewhat more acute course.

Another interesting fact was brought out in the bacteriologic
inestigation of these cases — namely, that the cultural character-
istics of the B. hovisepticus are not constant. Even with organisms
of approximately equal virulence the growths on and in media vary
considerably (these are perhaps most marked in bouillon as the
accompanjdng table will show), and with races of unequal virulence
the differences are still greater.

32

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33

A fuller discussion of the variations of B. hovisepticus, especially
as regards pathogenicity, must be left for further study. From the
few facts at the present time available we are not willing to draw
conclusions.

REFERENCES TO LITERATURE.

DiEKERHOFF. Lehrbuch der speciellen Pathologie und Therapie. Berlin,
1892.

ScHXEiDMiJHL. Vcrgleichende Pathologie und Therapie. Leipzig, 1898.

MooEE. The Pathology and Differential Diagnosis of Infectious Diseases of
Animals. Ithaca, N. Y., 1902.

WooLLEY and Jobling. Hemorrhagic Septicaemia in Animals in the Philip-
pine Islands. Bulletin No. 9, Biological Laboratory, Bureau of Gov-
ernment Laboratories. Manila, 1904.

WooLLEY. Pulmonary Lesions Produced by the Bacillus of Hemorrhagic
Septicsemia. Bulletin No. 12, Ibid. Manila, P. L, 1904.

23394 3

Part III.
REPORT ON PINTO (PAnO BLANCO).

By Paul G. Woolley, M. D., Director of the Berum Laboratory.

Under the terms paiio bianco, pinta, pinto, carate, mal pintado,
mal de los pintos, mal del pinto, peint, cute, cativi, quirica, pannus,
carateus, and the spotted disease of Central America, is included
a group dermatomycoses, characterized by peculiar nonpigmented
patches on the skin, in the scales from which hyphaj or spores or
both, of a mold-like fungus are found, which resemble in some
cases Penicilium, in others Aspergillus, in still others Monilia.

Heretofore this epiphytic disorder has been reported from Mexico
and Central and South America; another disease resembling it in
some respects has been observed by Legrain in North Africa, and
by Sandwith in Egypt, but, so far as I know, no previous report
has come from the Philippine Islands.

The case which I wish to record is not the only one which I have
seen in Manila, but it is the only one from which I have been able
to obtain specimens for examination. All of the affected persons
whom I have noticed have shown only the white variety, of which
the following case is an example.

The history of the case is as follows : The patient was a Filipino
laundryman, 15 years old, and in good health. There was no
similar disease in any of his immediate family.

Upon inspection it was noticed that there were pinkish white
patches, irregular in size and shape, on the ankles, dorsa of the
feet, shins, knees, elbows, hands, wrists, and one on the right
shoulder. This last-mentioned lesion the boy saj-s was the one first
noticed. The largest ones were over the external malleoli of the

35

36

ankles. These, the boy says^ appeared after the one on the shoulder.
The patches on the knees and elbows occurred later. None of
these patches were of the same shape or size, nor were they definitely
defined, but they shaded from their clear Avhite centers to the
normal brown of the skin. Neither weve the lines of extension
regular, so that the outlines of the patches were irregular and
crenated. About the larger areas were smaller ones, some barely
visible and of a faint pinkish white or very light-brown color.
. On palpation it was evident that the skin over the larger patches
was slightly rougher than the normal and that it felt somewhat
thicker. The palpating finger could detect no abnormal variation
in the covering of the smaller spots. There was but a minimum
amount of scaling, and there was some itching.

The rate of extension had been extremely slow, for in three years
the largest patch had a diameter of but 7 by 5 centimeters.

When asked regarding the cause, the boy said that the first spot
caine from carrying laundry baskets on his shoulder, and that the
other ones followed traumata of one kind or another. There were
no lesions on the palms of the hands or soles of the feet.

From one of the larger lesions on the ankle scrapings were made
and examined in a solution of caustic potash (25 per cent) . Among
the epithelial cells branching, segmented hyph« were seen forming
a coarse mesh work. The mycelium was somewhat finer than that of
Trichophyton; it Avas in general evenly refrangent, but in places
beaded or granular. The spores were darker in color than the rest
of the organism and less refractile. An occasional fructification
was found in the smears, and in these the arrangement of the
spores was like that of Penicilium. When treated with dilute
fuchsin the spores were stained a very deep red. The hyphse
showed an inner segmented arrangement with a continuous inclosing
capsule.

There was nothing in any of the preparations to suggest the
description of Gastambide. The mycelial filaments were usually
long, branched, and terminated in a bunch of spores. The descrip-
tion given by Montoya y Plores seems to apply more accurately
to the fungus of this case.

There can be no doubt of the nature of the disease. The clear
white spots with almost normal looking skin can be confused with
no other skin affection with which I am acquainted. Diseases
caused by trichophytons are extremely common in Manila and are

37

known generally as "dhobie itch," which is so common in the natives
that in thirty cases of skin disease taken at random in Bilibid Prison
twenty-four showed trichophyton filaments in cavistic potash
preparations.

It is possible that a brown pinta might be confused with pityriasis
versicolor should the small patches occur on the face where it -is
said that the latter may occur. However, in the present case the
clear white color of the irregular patches, the presence of sensation
and of itching, together with microscopic findings, are enough to
assure a correct diagnosis.

REFERENCES TO LITERATURE.

Manson. Tropical Diseases. London, 1903.
SCHEUBE. Krankheiten der Warmen Lander, 1900.

Part IV.

NOTES ON ANALYSIS OF THE WATER FROM
THE MANILA WATER SUPPLY.

By Charles L. Bliss, B. S., Chemist.

,1 >

In December, 1903, it was suggested by the Superintendent of
Government Laboratories and the Director of the Biological Labor-
atory that a systematic investigation, botli from a chemical and
bacteriological standpoint, be made of the water supplied the city
of Manila. After careful consideration it was decided that this
investigaton should consist of the usual sanitary chemical analysis
and a count of bacteria, these to be made at regular intervals of
one week and to extend over a period of several months. Later it
was decided to examine the water for the presence of amebas and
also to extend the investigation so as to include samples taken from
the course of the watershed.

There were several reasons why such examinations were desirable :

(1) Up to that time no chemical analysis of the city water had
been made at the laboratory, and frequent requests for statements
in regard to the water had to be denied.

(2) It was supposed the water would vary considerably from
time to time — ^not only from one season to another, but even from
day to day. It seemed very probable that during the rainy season
especially it would be subject to great changes within the course of
a few hours, and that the results would be markedly different from
the ones obtained during the dry, hot months preceding. This
seemed all the more probable if any conclusions can be reached
from the appearance of the water on different days. While ordi-

39

40

narily it is quite clear^ a few hours after a rain it shows turbidity,
which is very marked after a heavy storm. The watershed contains
a quantity of suspended matter in the form of a fine silt, most of
which gradually deposits when it is allowed to stand for a short
time. This would naturally lead to the supposition that a variation
could also be expected in the soluble constituents and in the number
of bacteria.

(3) Although numerous bacteriological examinations have been
made of the city water since the organization of the Biological
Laboratory, these had not previously been made at regular intervals,
but only at random as opportunity offered; moreover, they were
undertaken for different purposes. At times merely a count of
bacteria was needed; at others, a search for certain specific organ-
isms was undertaken, so that no continuous record was made of
the number of organisms found in the water.

(4) Amebas had been frequently found in the water in past
times,^ but it was not definitely known whether or not they were
always present. With the prevalence of amebic dysentery in the
city it was especially desirable to know whether these organisms
were constantly present in the water.

The investigation was begun on December 14, 1903, and con-
tinued during seven months. It extended over the greater por-
tion of the cooler weather, the hot season, and the beginning of
the rainy season. At first weekly examinations were made; but
after three months, there having been but little variation, examina-
tions were made every alternate week. All samples were taken
from a tap in the laboratory except on the following days : Feb-
ruary 23, from El Deposito; February 29, from the Mariquina
Eiver at Santolan, and March 7, four samples from the Mariquina
Kiver (the watershed) at different points.

The taking of the samjjles (with the exception of those from
El Deposito and Mariquina Eiver), the methods of manipulation,
the apparatus and chemicals were as nearly uniform as possible
throughout the entire period, Monday of each week being selected
as the day for doing this work. The last sample was analyzed on

^ Dr. Strong called attention to the presence of amebas in the water
and to its unfitness for drinking purposes in his annual report as Director
of the Biological Laboratory in 1902 and 1903. See Report of the
Superintendent of Government Laboratories for 1902 and 1903.

41

July 15, for the reason that there had been an excessively heavy
rainfall on the four days preceding, during twenty-seven hours of
which time 17-^ inches of rain had fallen, the city and outlying
districts having been completely under water from the evening of
the 12th till a day or so later. A few weeks earlier an examina-
tion made eight or nine days after a rather severe typhoon showed
results very nearly the same as those which had been obtained
throughout the series ; it was therefore thought best to take a sample
very soon after the flood, rather than wait until the following week.
It might be remarked that even in this instance there was but
very little variation from the usual results.

Two samples were taken from one of the laboratory taps each
Monday morning, after the water had been allowed to run for at
least one-half hour; one was retained for chemical analysis, the
other, with the usual precautions to prevent contamination, was
sent to the Biological Laboratory.^ Both examinations were begun
immediately. The sample for chemical analysis was collected in
a 3-liter, glass-stoppered bottle, which was first rinsed thoroughly,
filled and emptied two or three times, and then filled to the neck;
the stopper was well rinsed and immediate^ inserted. This bottle
was used for no other purpose throughout the series. The deter-
minations of nitrites, nitrates, ammonia, and oxygen consumed
were started at once so as to obtain results representing the true
condition of the water before any changes due to oxidation, reduc-
tion, or bacterial action could vitiate it, all necessary precau-
tions to prevent contamination by laboratory fumes being taken.
As the results in chlorine and hardness would not be affected
by any changes which might take place in the water, and as the
residues would be but very little if at all altered, these determina-
tions were deferred until opportunity to make them was at hand;
but in every instance they were undertaken before the end of
the week. In order to represent the water as it actually came from
the pipes, all analyses were made with the unfiltered liquid.

The chemical analysis consisted of the determination of total
residue; fixed residue; loss on ignition; nitrogen in the forms of

^ The counting of the bacteria was done by Mr. Clegg, of the Biological
Laboratory, and the determination of the presence of amebas was also done
by Mr. Clegg in conjunction with Dr. Musgrave, who has compiled his
results in Bulletin No. 18, Bureau of Government Laboratories, Biological
Laboratory.

42

nitrites^ nitrates, free and albuminoid ammonia ; oxygen consumed ;
chlorine; also hardness. As some of the methods of manipulation
were different from those usually employed, a brief description is
given.

Total residue was determined in a platinum dish easily holding
100 cubic centimeters, by evaporation on the water bath, and
heating for thirty minutes at 95° after drying. The dish, after
the weighing of the total residue, was heated uniformly to low
redness for three or four minutes. It was then placed in the
desiccator and weighed as soon as cold. There was practically
no change in the appearance in any sample on heating; at most only
a very slight darkening and but little odor were perceptible,
indicating the presence of only small amounts of organic matter.
The loss on ignition was therefore due largely to decomposition of
carbonate, as the heat was not sufficient to volatilize any chlorides.

Nitrites. — Tlie reagents Avere prepared as follows :

{a) Eight grams of naphthylamine hydrochloride were dissolved
in water, 8 cubic centimeters of concentrated hydrochloric acid
added, and the solution diluted to one liter.

(6) Sulphanilic acid, a saturated solution in water containing
5 per cent concentrated hydrochloric acid. The test Avas made by
placing 50 cubic centimeters of the water in a Nessler tube, adding
1 cubic centimeter of each of the above solutions, and mixing well
by gently shaking. After thirty minutes the color was noted.
In no instance did the depth of color indicate more than a very
faint trace of nitrites, and very frequently none developed. There-
fore a quantitative estimation could not be made.

Nitrates. — The aluminum reduction method was employed.
After a few hours, when the reaction Avas completed, the ammonia
formed, together with the free ammonia originally present in the
Avater was estimated directly by the Kessler process; the free
ammonia (determined in another portion of the sample) Avas
deducted, the remainder being the ammonia formed from the
nitrates. In the present series the determination of nitrates was
begun within a few minutes after the sample was received, and
the IvTesslerizing was done on the following day. If nitrites are
present in appreciable amount an allowance should be made for
them.

A little more than 50 cubic centimeters of the sample Avere placed

43

in a 250 cubic centimeter glass-stoppered bottle ; 2 cubic centimeters
of sodium hydroxide of 33 per cent strength and 2 grams of
aluminum filings were added, and the loosely stoppered bottle
allowed to stand at room temperature until the next day. The
solution was then filtered, with all precaution, into a tube, filling it
to the 50 cubic centimeter mark. This solution was then Nessler-
ized, the necessary correction for free ammonia being made.

Free ammonia. — A round-bottom flask of 1 liter capacity with
short neck was connected with a condenser 1 meter in length, the
ISTessler tube being slipped over the other end. On the day the
sample was received the distilling flask was rinsed with distilled
water; about 200 cubic centimeters of a solution of distilled water
containing 1 gram of sodium carbonate were then added, and the
greater part of the water distilled off, until the apparatus was free
from ammonia. After cooling, 500 cubic centimeters of the sample
were added to the residue, and the distillation continued at such
a rate that a i^essler tube was filled to the 50 cubic centimeter
mark in about ten minutes. Three tubes of 50 cubic centimeters
each were collected and Nesslerized. As a matter of fact one
tube would usually have been sufficient, at most two ; for the third
one never showed more than a slight trace of ammonia, and often
none at all.

Albuminoid ammonia. — Fifty cubic centimeters of alkaline per-
manganate (8 grams of permanganate, 200 grams of potassium
hydroxide, and 1,100 cubic centimeters of water, evaporated to
1 liter) were now added to the contents of the flask and distilla-
tion was continued, four tubes of 50 cubic centimeters each being
collected. The distilling apparatus was used for no other purpose
throughout the series, and it was always well protected from fumes.

Nesshrizing. — The Nessler solution was prepared according to
the usual method and the standard solution of ammonium chloride
contained (0.03812) gram of pure ammonium chloride in one liter.
One cubic centimeter represents 0.00001 gram of nitrogen.

The distilled water in the laboratory was found to be free from
even the slightest perceptible trace of ammonia ; it was tested
each time. A number of Kessler tubes were thoroughly rinsed
with this ammonia-free water, then filled to the 50 cubic centimeter
mark; portions of the standard ammonium chloride solution were
measured in from a normal capillary pipette reading accurately

44

in hundredths of a cubic centimeter. . The amounts used were 0.2,
0.4, 0.6, 0.8, 1, 1.2, 1.5, 2, and 2.5 cubic centimeters. Two cubic
centimeters of the ISTessler reagent were added to each, and also
to the tubes containing the ammonia (free, albuminoid, and that
from the nitrates). After allowing thirty minutes for develop-
ment of the color, the comparisons were made. The results were
expressed in terms of nitrogen per million.

Oxygen consumed. — The solution of pure potassium perman-
ganate contained exactly 0.3953 grams in 1 liter. One cubic centi-
meter represented 0.0001 gram available oxygen, and the solution
of pure ammonium oxalate was of exactly equivalent strength.
These two solutions were kept in a dark closet and were standardized
from time to time.

One hundred cubic centimeters of the water were measured into
a 200 cubic centimeter Erlenmeyer flask from a pipette; 2 cubic
centimeters of pure concentrated sulphuric acid were added, and
then 10 cubic centimeters of the permanganate solution; the flask
was then suspended in a boiling-water bath for thirty minutes.
It was then taken out and 10 cubic centimeters of the oxalate
solution were immediately added. The solution became colorless
within a minute or so, and was then titrated with the permanganate
till a faint pink color appeared, which remained permanent for a
few moments. This determination was made in duplicate, two
flasks "being carried through at the same time. In no instance
did the two titrations vary more than 0.05 cubic centimeters, and
usually they were identical. For convenience, the titrations
were made with a very narrow pipette graduated in 0.05 cubic
centimeters. Eesults were expressed as parts of oxygen consumed
per million.

Chlorine. — A solution of pure, recrystallized and dried silver
nitrate was made containing 4.7940 grams in 1 liter. One cubic
centimeter represented 0.001 gram of chlorine. A 5-per cent
solution of potassium chromate freed from chlorides by precipitat-
ing with silver nitrate was used. The chlorine determinations
were made in duplicate. Two portions of 250 cubic centimeters
each were measured into casseroles of about 300 cubic centimeters
capacity, carefully evaporated to about 50 cubic centimeters, 1
cubic centimeter of the chromate solution added, and the mixture
titrated with the silver solution. By using one as a comparison

45

the first change in color from a pure to a reddish yellow was
noted very sharj^ly, and the two results never varied by more than
0.05 cubic centimeter; usually they were identical. A pipette
similar to the one employed in the titration with permanganate
was used. Eesults were expressed in terms of chlorine per million.
Hardness. — A solution of pure calcium chloride was diluted so
that 1 cubic centimeter contained 0.0003218 grams, or represented
the equivalent of 0.0003 gram of calcium carbonate. Fifty cubic
centimeters of this solution should require exactly 14.35 cubic
centimeters of standard diluted soap solution in order to form a
lather which covered the surface of the liquid and persisted for
five minutes. The soap solution was standardized each time and
the proper value obtained, 50 cubic centimeters of the above calcium
chloride being used for this purpose. This was compared with
50 cubic centimeters of the sample treated in the same manner,
and the degree of hardness determined by reference to published
tables. In the two or three instances, when the results varied from
those usually obtained, and in all cases where any doubt as to the
exact end-point existed the determinations were repeated.

Analyses of the Manila ivater supply.
[Results are given in parts per million.]

Labo-
ratory
No.

Location.

Date.

Total

resi-
due.

Fixed
residue.

Loss
on
igni-
tion.

Nitrites.

Ni- .
trates.

Free
am-
monia.

1739
1745
1756
1780
1781
1803
1821
1840
1880
1922
1944
1996
2025
2026
2027
2028
2069
2157
2225
2303
2352
2385
2434
2464
2487
2511

Dec. 14,1903
Dec. 21,1903
Dec. 28,1903
Jan. 4, 1904
Jan. 11,1904
Jan. 18,1904
Jan. 25,1904
Feb. 1, 1904
Feb. 8, 1904
Feb. 15,1904
Feb. 23,1904
Feb. 29,1904
Mar. 7, 1904
.do

220
181
188
188
191
179
176
178
162
168
168
164
160
153
174
173
167
173
174
165
180
169
196
174
178
191

190
148
152
142
160
148
152
152
142
152
145
138
134
127
147
145
138
147
138
136
145
135
150
138
150
159

30
33
36
46
31
31
24
26
20
16
23
26
26
26
27
28
29
26
36
29
35
34
46
36
28
32

0. 150
.271
.100
.112
.080
.110
.198
Trace.
.100
.138
.274
.292
.124
.139
.112
.136
.220
.220
.158
.200
.158
.120
.360

0. 0079

.0049

.000

.008

.002

.006

.002
Trace.
Trace.

.002

.006

.028

.036

.021

.028

.024

Trace.

Trace.

.002
Trace.

.004
Trace.
Trace.
Trace.
Trace.
Trace.

o"

0
0
0
0

<■' 0

0
0
0
0
Trace.
0

Trace.
Trace.

(1)
(1)
(^)

Santolan

Mariquina River _
do

do -

do

do

do

Mar. 14,1904
Mar. 28,1904
Apr. 11,1904
Apr. 2.5,1904
May 9, 1904
May 23,1904
June 6,1904
June 20, 1904
July 5, 1904
July 15, 1904

0

0 .360
0 .220

46

Analyses of the Manila water supply — Continued.
[Results are given in parts per million.]

Labo-
ratory
No.

1739

1745
1756

1780

1781
1803
1821
1840

1880
1922
1944
1996
2025

2026
2027
2028
2069
2157
2225
2303
2352
2385
2434

2487
2511

Locati >n.

Deposito

Santolan

M a r i q u i n a
River.

do
do
do

Albu-
min
am-
monia.

.073
.031

.034
.038
.044
.048

.048
.044
.052
.086
.100

.080
.062
.060
.048
.052
.062
.048
.064
.042
.074

.040

.050
.068

Oxy-
gen.

1.90

1.85
.875

.85
.65
.65
.90

.90

.95

.95

1.25

1.60

1.50
1.07
1.35
1.00
.82
1.07
1.20
1.20
1.05
1.60

1.70
2.20

Chlo-
rine.

2.13

2.23
2.60

2.40

3.00
3.04
3.00
3.00

2.60
3.20
3.30
3.40
3.60

3.20
3.60
3.60
3.88
4.14
3.80
4.40
4.20
4.40
4.20

2.80
3.16

Hard-
ness.

85.7

85.7
90.0

94.3

101.5
94.3
87.1
95.0

104.0
95.0

109.0
97.0
97.0

93.2
93.2
94.8
95.0
101.8
97.8
95.0
98.0
92.0
91.0

83.0

71.4

58.8

Bacte-
ria.

550
600

250
200
200
150

125
150
120
112

208
105
267
120
120
108
125
175
130
206

450

Ame-
bas.

0

Remarks.

Heavy rain dur-
ing few days
preceding;
water very
turbid.
Do.
Water distinctly

turbid.
Water slightly
turbid.
Do.
Do.
Do.
Water almost
perfectly clear.
Do.
Do.

All contained
deposit, appar-
ently organic
matter and
silt. Determi-
nations of
residue were
made with fil-
tered water.
Each gave
sliglit dark
color and odor
on ignition.

Do.

Do.

Do.

Heavy rains just
previous; wa-
ter turbid;
slight sedi-
ment. Slight-
ly dark color
and odor on
ignition.

No rain imme-
diately pre-
ceding.

Very turbid.
Heavy rains.

Very turbid .
(Flood.)

1 Very faint trace.

2 Faint trace.

3 Distinct trace.

*Not sufficient sample. A sample taken following week, bacteria, 100; amebas, 0.
5 Rapid growth over surface prevented count.

The counts of bacteria and determinations of the presence of amebas were made by
Dr. W. E. Musgrave and Mr. M. T. Clegg, both of the Biological Laboratory .

47

On examining the above table the following maxima and minima
appear during the period covered by the report :

Total residue

PMxed residue

Loss on ignition

Nitrites (N)

Nitrates (N)

Free ammonia (N)

Albuminoid ammonia (N)
Oxygen consumed (O) .__

Chlorine (CI)

Hardness

Minimum.

Maximum.

153

220

127

190

16

46

0

Trace.

Trace.

.36

0

.036

.031

.100

.65

2.20

2.13

4.40

58.8

109

As will be seen despite the great variation in the weather condi-
tions the differences in analytical results were not very great. At
times the chlorine ran as high as 4.40 indicating some contamina-
tion, but these maxima were only transitory. However, a water
may show a very high degree of purity, so far as this can be deter-
mined by chemical analysis, and yet be unfit for drinking purposes
because of either the large number of micro-organisms which it
contains or because of their nature; so that it maj^ be possible to
convey the etiological factor of typhoid fever, cholera, dysentery,
etc., by a drinking water which chemically would be pronounced
unobjectionable. The chemical analysis may indicate a probable
pollution with sewage or with other matter which may be suspicious
or dangerous at all times, and it may condemn such a water. How-
ever, in the case of the Manila supply the long series of analyses
gave such results that no one would be justified, even at the worst,
in stating from a chemical standpoint that this water was either
injurious or deleterious to the public health. For this reason, as
has been repeatedly pointed out by others, a bacteriological examina-
tion is essential.

However, in glancing over the number of bacteria it will be seen
that the maximum, on December 28, was 600, and this large number
was quite unusual, the average being below 250 organisms to 1
cubic centimeter and in many cases even below 150. Bacteriolog-
ically, therefore, the water may not be regarded as very suspicious,
especially since the general series of determinations did not demon-
strate pathogenic organisms to be present, and indeed typhoid fever
is of but rare occurrence in Manila. During the cholera epidemic
it does not seem likely that any of the cases could have been referred
to direct infection from the Manila water suj)ply.

48

A factor however, which, apart from the chemical and bacterio-
logical analysis, is the most important, is shown in the last column,
where it is demonstrated that amebas, whether pathogenic or not,
are almost constantly present in the water supply, and Dr. Mns-
grave and Mr. Clegg, of the Biological Laboratory, in Bulletin
No. 18, "Amebas: Their Cultivation and Etiologic Significance,"
have shown that amebic dysentery can be sometimes produced in
monkeys by cultures made from the water supply. Neither a
chemical analysis nor a bacteria count will demonstrate the presence
of these dangerous factors of disease, and consequently, in the
Tropics at least, if we wish to obtain a fair idea of the condition of
the water, a determination of the presence or absence of amebas is
necessary.

The ordinary sanitary analysis does not include an examination
for substances which in themselves are injurious. The quantities
of ammonia, nitrate, chlorine, etc., found in an ordinary water are
harmless — they are simply indicative of a possible pollution, but
before judgment can be passed in regard to the sanitary analysis,
the source of the water, the geological conformation of its sur-
roundings, and so forth, must be taken into consideration. An
amount of chlorine, for example, which would be perfectly normal
in water from one locality might indicate contamination with
sewage in that from another. The total residue in this series was
always below 200 parts per million excepting on one occasion
which was after a heavy rain.

Very little darkening of the residue on heating was noted in any
sample; sometimes there was' .none, so that in this respect no
criticism can be made of the water. The presence of nitrites in
measurable quantity is sufficient ground for condemning a water as
a rule, for nitrites indicate bacterial action. However, in this
water nitrites were frequently absent altogether, and only on one
occasion were there more than a trace, this occurring immediately
after a heavy rain.

The amount of nitrates was always very low. The highest
amount of free ammonia found was 0.008 per million, excepting
in five samples . taken from the Mariquina Eiver itself where the
results varied from 0.0021 to (0.036), so that by the time the water
from the river reaches Manila the ammonia has largely become
oxidized to nitrate. This is borne out by the fact that the amount
of nitrate in those samples which were taken on March 7 was

49

lower than the usual quantity obtained. Albuminoid ammonia, as
a rule, was also quite low. None of these factors, therefore, would
indicate a contamination of the river water.

The same may be said of the amount of ox3^gen consumed. No
figure exceeding 2 was found excepting in one instance, and that was
three days after the great flood in July. The amount of oxygen
consumed seemed to be greater immediately after heavy rains.

The low results obtained in the investigation of chlorine are
favorable indications as to the quality of the water. One would
expect rather higher chlorine values in the waters of the Philip-
pine Islands owing to the proximity to the sea, but this is not as
a rule the case, as analyses of waters from other localities have
demonstrated.

After heavy rains more or less insoluble suspended matter, both
inorganic and organic, is rinsed into the suppty. However the
turbidity of the Manila water is generally due to a very fine silt
which has its rise in some of the clay beds at the source of the
river, and is for this reason harmless. Therefore the analyses made
in the Chemical Laboratory show that the water supply of the city of
Manila is of a very good quality, but the constant presence of
amebas, as demonstrated in the Biological Laboratory, render the
water unsafe for drinking ]iur]~)oses unless it is boiled.
23394 4

Part V.

FRAMBOESIA: ITS OCCURRENCE IN NATIVES
OF THE PHILIPPINE ISLANDS.

By Paul G. Woolley, M. D., Director of the Serum Laboratory.

Qy

In a recent visit to Benguet Province, in the central part of
northern Luzon, I became interested in a peculiar disease which
was called "Lepra" by the native Igorrotes, this term having been
taught them by the Spaniards. Inasmuch as I was entirely unpre-
pared for a complete study, and because it is extremely difficult to
obtain any history or to make complete examinations of these
people, the following report will be meager, but it is interesting,
since I feel certain that the disease is one closely related to if not
identical with framboesia.

In Baguio, Benguet, I saw, with Dr. Thomas of the Civil
Sanitarium, two cases; one, a woman aged about 35 years, and her
son, the latter of some 15 years, both of whom presented small rasp-
berry growths upon the face. In the mother, these were situated at
the corners of the mouth; in the son, in the nasolabial folds, and
they much resembled the growth pictured in the ISTew Sydenham
Society's Atlas (Fasc. XIV, PI. B. fig. 8). Both' of these persons
showed pigmented scars on the neck and face. At a later time I
searched for these two people in their native town of Agno, Beu-
guet, but was unable to find them.

The case from which I procured the tissue which I will describe
below, I saw one morning as Mr. Barron — the sanitary inspector of
the province — and I were returning from a long trip in the
mountains. As we stopped to rest at a little native village, we

51

52

noticed that several of the children had peculiar looking, sluggish
ulcers on their legs, necks, or bodies, and also showed signs of
considerable anemia. We asked concerning this affection and were
told that the Spanish called it "Lepra," and that many people had
it. This was all we could learn, except that the sores were not
painful, though they showed the effects of scratching, and that
they eventually healed. One grown person, a woman, was found
who had similar lesions on her neck. Whether she had others
elsewhere we were unable to discover. One lesion on the neck
(see Sydenliam Society's Atlas, Fasc. XIV, PI. LXXXY, upper
left-hand corner of lower figure) was a shallow ulcer with a firm
grey base, a well-defined, firm margin, a slight yellowish secretion,
not surrounded by any appreciable indviration, not painful, and
situated on the neck just below the angle of the jaw. This was
excised and preserved in commercial alcohol. Xear it were some
pigmented areas, somewhat darker than the normal skin^ not appre-
ciably thickened, and possessed of sensation, and which the woman
said were at the sites of healed ulcers similar to the ones we saw.
(New Syden. Soc. Atlas, PI. LXXXXV.) An ijifant which the
woman carried had similar lesions on its legs, face, and neck.
Another child of the same woman, which, however, we did not see,
was said to be afflicted in the same manner.

The gross appearance of the lesions would lead one to think of
leprosy, tuberculosis, epitheliomia, syphilis, or yaws. Leprosy I
think may be excluded. There were no cases of outspoken leprosy
among the persons of the pueblo in which these persons lived.
There were no anesthesias or leucoplakias in any of the cases
examined. The histologic examination was negative. Tuberculosis
could be excluded since neither the lesions nor the scars had the
classic appearance of lupus, nor was the histologic evidence suf-
ficient to support such a diagnosis. Epithelioma could only be
excluded by microscopic examination. All in all the cases seemed
most like syphilis. None of the people of the pueblo showed out-
spoken signs of this disease, although, as stated, a complete examina-
tion could not be obtained. The inhabitants of this region generally
are rigidly moral and rigorous punishment is inflicted upon any
who overstep the bounds. But while they live morally clean lives,
their physical surroundings are filthy, which may account for the
modification of the frambcesial lesion and the predominance of

53

infection. Treatment, of course, has not been tried, so one can not
say what effect mercury or iodides might have, but from the
evidence that I have at present I incline to the idea that the cases
seen were examples, not of syphilis, but of framboesia.

,The tissue was embedded in celloidin, and the sections which
were made later were stained with hsematoxylin and eosin, Unna's
polychrome blue. Gram's stain, and by the tubercle bacillus
methods. Microscopic examination showed that the lesion con-
sisted of a marked hyperplastic acanthosis, with a round-celled
infiltration of the underlying, and especially of the perivascular,
connective tissues. The first impression was that one was dealing
with an epithelioma, but closer examination showed this to be
a delusion. The acanthus layer of the skin was thickened and
prolonged in strands and columns of bizarre shapes. In many
places in this hyperplastic epithelium there were larger or smaller
islands of connective tissue, each, apparently, representing the path
of a blood vessel. In the centers of such areas and about the vessel
were collections of small round and plasma cells, but this small-
celled infiltration was most marked in the larger strands of the sub-
malpighian connective tissue. Here the increase of these formative
elements was remarkably prominent, and there was throughout the
sections the same perivascular arrangement. Within these areas
there were occasional leucocytes; there were also fibroblasts in
varying stages of development, and a number of plasma cells were
present within the round cell accumulations. At the site of the
ulceration the structure of the lesion was modified by the destructive
process. Here all the layers were invaded by a multitude of
polymorphonuclear leucocytes, the blood vessels were widely dilated,
and there was a certain amount of superficial degeneration, but the
structure of the lesion in the not degenerated parts was still per-
ceptible. So far as the arrangement of the layers of the skin was
concerned there was no distortion. In many parts of the sections
a peculiar appearance was seen which gave one the same impression
as that produced by the scales of a fish, or by the overlapping of
shingles upon the roof of a house. This was apparently due to
the fact that certain of the acanthus cells took a more intense stain
on one side. In this phenomenon (seen best in polychrome blue
specimens), the nucleus did not participate. In none of the sec-
tions, and these included the whole of the lesion studied, were there

54

any giant cells, tubercle of lepra bacilli, and no evidence of cell
inclusions was seen.

As for the occurrence of such lesions in framboesia, and their
distribjition, little can be said excepting as quotations from authors
who have had considerable experience with the disease. Manson ■
says, in discussing the distribution of the yaws, that they may be.
scattered over the entire body, or the crop may be limited to one or
two growths, or they may be confined to a circumscribed region of
the skin. Moreover, there may be successive crops evolved, especially
when the person affected is debilitated.' Morris remarks that the
disease in adults is more chronic than in children. When the yaw
develops normally it does not ulcerate, but Manson says that the
tumors instead of being absorbed, may break down and ulcerate,
the ulceration usually; being confined to the yaw itself, although it
may go deeper and give rise to extensive sores. With the develop-
ment of the deeper and more extensive forms of ulceration, the
typical lesion of framboesia may disappear for a time, or perhaps
permanently. If such is the case, the ulcerations are said not to
be infective and to not communicate the disease, although they may
persist for years. NichoUs, quoted by Mason, states that ulcera-
tion occurs in about 8 per cent of the cases. There is no histologic
description of the . variety of the lesion which I have encountered
to which I can refer, though f rom Unna's and Charlouis's descrip-
tion of the typical yaw it is but a modification of the latter. These
writers speak of the true yaw as a cutaneous plasmoma complicated
by epithelial hyperplasia and hyperkeratosis. Except that the lesion
described above is not raised it certainly corresponds in many
details with Unna's and Charlouis's description.

REFERENCES TO LITERATURE.

Manson. Tropical Diseases. London, 1903.

Morris. Diseases of the Skin. Philadelphia, 1898.

Hyde. Diseases of the Skin. Philadelphia, 1900.

Unna. Hautkrankheiten. Berlin, 1894.

ScHEUBE. Die Krankheiten der warmen Lander. Jena, 1900.

SECTION STAINED. WITH H/EMATOXYLIN AND EOSIN. (ZEISS COMP. OC. 6, OBJ. A. A.)

This shows the hyperplasia of the acanthus layer of the skin, the dilated blood vessels,
and the perivascular accumulations of small round and plasma cells. Photomicrograph
by Martin.

/

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