576.809.429+616.95+619.

31
ENZOOTIC HEPATITIS OR RIFT VALLEY FEVER.
VIRUSDISEASE
UNDESCRIBED
,4~ CATTLE
OF SHEEP
AND MAN FROM EASTAFRICA.

R. DAUBSEYand J. R. HUDSON,
Division of Veterinary Research,
Kenya Colony.
With an account of an experimental inoczllation of man by
P. C. GARNHAM, Xedieal Department.

(PLATES
XL1.-XLIII.)

INJ u l y 1930, an application was received for the assistance of the

Research Division in diagnosing a disease that was causing a heavy
mortality in newly-born lambs on a farm in the Rift Valley. The
farm i n question is situated a t a n altitude of 5500 to 6000 feet, and
extends northwards from the shore of Lake Naivasha. It consists
of about 30,000 acres of open bush country, and has carried Meriiio
sheep successfully for a number of years.
Normally, lambing takes place on this farm in October and
November; but on this occasion it was decided to lamb during July
and August, which are usually dry months. The peculiar climatic
conditions of East Africa make it a matter of some difficulty to
choose the most suitable periods for lambing. The choice of a season
has been narrowed down by experience to the October to November
period, which is just before the short rains, or to the month of May
just after the long rains. Further experience is leading to a general
adoption of the October to November lambing season as being more
suitable, since it has been found that lambs born during May suffer
much more heavily from parasitic infestations than those born in the
October to November season. I n deciding to lamb in July and August
this year the manager of this farm was no doubt largely influenced
by the unusual rainfall, which had been sufficient to ensure good
grazing throughout the dry season, and by a fear that should the
short rains in November also prove to be unduly heavy the pastures
might become rank and unsuitable for young sheep. The mean annual
JOURN. OF PATH.-VOL. XXXIV. 545 2 0
546 R. DAUBNEY AND' /. R. HUDSON

rainfall in the Naivasha region is approximately 25 inches, distributed

as follows :-
Inches. Inches.
January . . , 0'73 July . . . . 1'75
February . . . 1'04 August. . . . 2-02
March . . . . 2'44 September . . . 1'27
April . . . . 4.89 October . . . 1-32
>fay . . . . 3'61 November . . . 2'38
June , , . . 2'65 December . . . 1'13
During the year 1930, over 45 inches of rain were recorded,
31.44 iiiches falling during the " long " rains of February to May and
8.54 inches during the '' short " rains from September to November.
Sheep on this farm have not in the past suffered severely from
diseases of major importance. Sheep pox makes an annual or biennial
appearance, and is controlled by vaccination. Heartwater has been
responsible for some loss in the past, particularly in the areas border-
ing on Lake Naivasha, but it is now almost completely eradicated
as a result of periodic dipping of the cattle on the farm. Ticks
are no longer abundant on the grazing. The commonest species
are Rhipicephalus evertsi and Hyalomma Egyptiunt. Arnblyomma
variegatum is now rarely seen, and the brown-tick group is practically
unrepresented. Bluetongue has always existed on this farm, and
is regularly seasonal in its occurrence. The first cases are usually
observed towards the end of the short rains. In a normal season the
disease has died out by the end of January, although in occasional
years a few cases also occur after the long rains. Owing to the
excessive rainfall in 1930, there was a widespread enzootic of blue-
tongue throughout the colony, and the disease was still existent as
late as July.
During the latter part of June, before lambing had commenced, a
number of abortions occurred in the ewe-flocks. At first it was
thought that these abortions were due to bluetongue; since on two
occasions blood, taken after ewes had aborted, produced bluetongue
on inoculation into susceptible sheep a t the laboratory. However,
other samples similarly collected failed to transmit bluetongue on
inoculation.
Shortly after lambing commenced, it was realised that some disease
was causing an unduly high mortality in the lambs of certain flocks.
A t the outset the disease appeared to be confined to the lambs of
one ewe-flock, and by 10th July, sixty lambs had died out of a total
of eighty born in this flock. At the time of death the lambs were
from three to seven days old, the majority about three days. The day
before death they were listless, disinclined to feed, and sank down
soon after being put on their feet. Diarrhoea was not a feature of
the condition, and the symptoms were not very definite. Affected
lambs rarely survived twenty-four hours after the first appearance
of symptoms.
 RIFT VALLEY FEVER 647

Coincident with the appearance of this disease in lambs there was

a marked rise in the mortality rate of ewes on the farm, and the
number of abortiois also increased. I n contradistinction t,o the
position with regard to the lamb disease, both mortality and abortion
were observed among ewes in practically all flocks on the farni.
The ewes were either found dead without having shown previous
symptoms, or were observed t o be ill for a few hours only before
death. I n some cases it was stated that they were noticed to
vomit in the evening and were found dead the next morning. When
observed to be ill, the sheep showed a thick muco-purulent nasal
discharge, and occasionally the stools would consist of almost pure
blood. They were not feeding, were disinclined to rise, and a t this
stage had a normal temperature. Ewes that aborted were not visibly
ill a t the time, with the exception of a few animals that developed
septic metritis or peritonitis. .
On the 27th July, a second visit was made to this farm. It was
found that the situation had become far more serious, and that lamb
disease had extended, or rather, that cases were now occurring in all
the flocks of lambing ewes. Abortions among the ewes had almost
ceased ; but the ewe mortality was now extremely heavy.
A fortnight later, when a third visit was made, the posit,ion was
much the same. The total mortality in lambs had by now reached
approximately 3,500, and upwards of 1,200 ewes liad died.
By this time, it was already known from experiments that the
disease in the lanibs was a hyperacute forin of that in the ewes, and
that abortion was a common sequel to recovery in pregnant animals.
It had been docidea to remove the ewes and the remaining lambs by
easy stages to a n adjoining farm on the edge of the Rift Valley, a t an
altitude of 7000 to 8500 feet. Some of the flocks were then on their
way to the higher farm, and both ewes and lambs were dying on the
road. After the arrival of the sheep a t the higher farm, deaths
continued for a further period of five or six days. From that time
onwards no fresh cases were recorded, and it is reasonable to suppose that
those animals that died during the first few days after the movement,
had been infected before leaving the lower altitude. As the records of
experiments will show, the disease continued on the lower farm for several
months, and in fact, occasional cases were observed as late as December.
Post-mortem examination.
The lesions encountered a t post-mortem examination are very
constant in young lambs, whether naturally or experimentally infected.
I n gdult sheep the lesions a t first appear to be quite,different from
those found in lambs; this applies more particularly to the liver
lesion. Further investigations have shown quite clearly t,hat this
apparent difference depends solely upon the acuteness of the process
and upon the rapidity with which the lesions develop.
548 R. Dd UBNE Y AND 3. R. HUDSON

Live?.. The most striking lesion in lambs is an extensive necrosis

of the liver. At the outset this is a focal process; but in a severe case
most of the liver tissue is affected. Usually, however, some part of
the surface of the organ can be found, which still indicates quite clearly
the focal nature of the lesion. The necrotic foci appear beneath the
capsule as small, white spots, up to about 1 millimetre in diameter.
In the neighbourhood of such small foci one commonly finds some
scattered pin-point subcapsular haemorrhages, reddish-black in colour.
The liver itself is not enlarged.
When a certain amount of normal tissue remains its colour is
usually relatively unchanged ; but occasionally the whole liver may
be light yellow (fig. '7). The necrotic foci are distributed more or less
evenly throughout the whole of the organ, and when they are sufficiently
numerous,coalesce to form a diffuse necrotic lesion. There is never
any sign of demarcation of large necrotic masses from surrounding
tissue.
On cutting the liver it is found that the lesions visible on the
surface extend throughout the substance of the organ. I n general,
the liver in acute cases shows a striking resemblance to that seen in
certain Sulmonella infections such as fowl typhoid, and to a less extent
paratyphoid of calves.
The acute liver lesion described above is only rarely seen in adult
sheep ; usually the changes are much less advanced. I n ewes dying of
the natural or experimentally-induced disease, the surface of tho liver
appears at first sight to be mottled with small subcapsular hzmorrhages,
which are visible as pin-head size reddish-brown spots. Actually this
red mottled appearance is due to the presence of innumerable ill-defined
necrotic foci, which are paler in colour than the normal tissue, and which
by contrast make the normal tissues appear congested or hzmorrhagic.
Although this red mottled appearance is characteristic of the liver
lesion in adult sheep, there is often a small area where the necrotic
foci are well-defined and readily recognizable.
On section the cut surface usually appears paler than normal;
distinct focal lesions, either of the congestive or necrotic type are not
usually visible to the naked eye. Increase in the size of the organ is
not noticeable, although it is occasionally engorged with blood.
Spleen. The spleen is not enlarged. Tumor splenis has been
observed only once in an experimental case in the course of some
two hundred post-mortem examinations. The constant lesion is the
presence of pin-point to pin-head size hsmorrhages beneath the capsule.
These haemorrhages are chiefly found towards the free border of the
organ. Capillary arborescence in the neighbourhood of the petechize
is of frequent occurrence. The pulp of the organ is not noticeably
affected. The condition of the spleen is an important diagnostic
feature, since it enables one to exclude Nairobi sheep disease, heart-
water and bluetongue from consideration as the cause of death.
 R l F T VALLEY FEVER 549

Kidney. The capsule of the kidney strips normally, exposing a

smooth cortical surface, which, however, usually shows a few small
ill-defined hsmorrhages or some capillary arborescence. On section,
both cortex and medulla not infrequently show a few radiat,ing lines
of congestion. Otherwise the tissue appears normal.
Lungs. The lungs are little changed. They are normal in size and
colour and do not show hsmorrhages. A fairly constant lesion is the
presence of small slightly raised areas on the surface of the organ.
These areas are irregularly oval i n shape, and may be up to an inch
in the larger diameter. They are a little paler than the surrounding
tissue. The bronchi and trachea show nothing unusual, although the
mucous membrane of the larynx may occasionally be markedly
congested.
Circulatory system. Subepicardial ecchymoses are common in the
region of the coronary grooves on the ventricles of the heart. I n
addition the left ventricle almost invariably shows fairly extensive
sub-endocardial extravasations. In the right heart, lesions are
extremely rare. It has been noticed when drawing blood for virus
that the jugular vein is always difficult to raise. The mesenteric and
omental vessels are always strongly injected. The injection is another
characteristic feature of the disease.
Xymphatic system. The mesenteric lymphatic glands are always
enlarged, and on section appear more moist than normal. In most
cases there are extensive haemorrhages in the cortex of these glands.
Alimentary tract. The lesions in the alimentary tract are vague
and subject to considerable variation. The mouth, pharynx and first
three compartments of the stomach show nothing unusual, The
mucous membrane of the abomasum may be faintly congested, or
niay on occasion show numerous pin-point hsemorrhages towards the
cardiac end.
As a rule the duodenum and jejunum are little changed, although
the mucous membrane may be catarrhal throughout the small intestine.
I n addition, the ileum is not infrequently the site of some diffuse
hyperaemia; and the cecum may show areas of congestion which
usually appear to be arranged roughly in transverse ridges, but
occasionally tako the form of longitudinal bands. The czcal lesion
may extend some little distance along the colon, which otherwise
shows no change. It should be mentioned that in some lambs the
cscum has been found impacted with a pultaceous mass resembling
a canine stool. Further, a few cases in which there is a marked
haemorrhagic enteritis, affecting chiefly the ileum and Iarge intestirie
and leading to considerable thickening of the mucous membrane, have
been seen in adult sheep. I n these cases the lumen of the colon is
impacted with chocolate-coloured faeces, " tarry " in consis teiice and
obviously containing a large amount of blood. These animals have
shown characteristic lesions of the subacute type, and the blood upon
.lOURN. OF PATH.-VOL. XXXXV. 2 0 2
550 R. DAUBNE Y A N D J. R. HUDSON

inoculation has produced the more usual form of the disease. It is

however, by no means certain that the " tarry " enteritis is not due
to some complicating infection.
There is a very remarkable tendency to rapid decomposition in the
carcases of animals that have died of the disease. Sheep that die in
the night are frequently too decomposed for post-mortem examination
by nine o'clock on the following morning. Doubtless this early
decomposition is related to a more or less complete failure of liver
function some time before death.
-iVemous system. The only macroscopic lesion found was injection
of the meningeal vessels.

Bacteriology.
From the nature of the lesions found in the first post-nzorta?ic
examiiiations in lambs, it was thought probable that sonie bacterial
infection, posaibly of the Salmonelln type, might be responsible for
the condition. Further, the constant presence of lesions in the
mesenteric glands, together with the absence of any infection of the
umbilical vessels, strongly indicated the alimentary canal as the
probable porlal of entry. On cultural examination, however, the
intestinal flora revealed nothing abnormal, nor was there any evidence
of excessive multiplication of B. coli or of the extension of faecal flora
to the small intestine.
Cultures were repeatedly made from the liver ancl rriesenteric
glands of naturally and experimentally infected sheep and lambs on
plates of plain agar, ancl on McConkey's, Conradi's and Endo's media.
KO non-lactose fermenting organisms were recoverecl, ancl with the
exception of stray contaminating bacteria the organs proved to be
sterile.
Transmission experiments.
Citrated blood taken from a naturally infected lamb in the field on
27th J ul y 1930 was inoculated intravenously on the following day into
sheep 33177. Unfortunately, through an error, the temperature of this
sheep TWS not taken on the day after inoculation ; but on the second
clay, although the animal showed a normal temperature it was obviously
sick. It mas lying down and was completely "off its feed." Respira-
tion was rapid, and there was a heavy blood-stained discharge from the
nose. The f me s were a little softer in consistency than normal. Blood
taken from this sheep on this day was inoculated intravenously into
two sheep, 3380 and 3383. Sheep 3380 showed a sharp rise in
temperature to 106.6" on the evening of the second day, but returned
to normal on the morning of the third day. The temperature of sheep
3383 rose to 106.6" on the morning of the second day, and reached
136.8" on the morning of the third day; by the fourth morning the
animal's temperature was normal (chart I).
 RZFT VALLE Y RE VER 551

On the day that blood was collected from the lamb, blood was also
taken from a typical case of the disease in ewes. This blood was
inoculated intravenously into sheep 3385. On the morning of the
second day this sheep had a temperature of 107", and its visible mucous
membranes were injected and dirty in appearance. I n the afternoon,
the animal's temperature fell t o lo@, and a profuse blood-stained

:'ol I i

DAYS AFTER INCCULATION.

OVlWB

3383.

CHARTI.
mucous discharge was present in the nostrils. This animal had a
further rise in temperature on the evening of the third and morning
of the fourth days, returning to normal on the fourth evening.
On the second day, blood from sheep 3385 was inoculated subcu-
taneously into sheep 3379 and intravenously into sheep 3381 and 3485.
652 I?. DAUBNEY AND /. R. BUDSON

Sheep 3379 reacted sharply, its temperature reaching 107.2" on the

evening of the first day after inoculation, and returning to normal on
the second evening (chart 11). Sheep 3381 reacted on the first clay,
arid had a normal temperature on the second morning. Sheep 3485
reacted on the first day, and died on the morning of the third day.
This animal was a native ewe that was lambing, probably somewhat;

1 a:

1 Q:

Wlhm lo.'
5379.

1EJ'

1 oa'

101'

ovim
3300

prematurely, on the third day, and death was no doubt accelerated by

a complicating peritonitis.
Further inoculations with the original material were also carried
out on 30th July 1930. Sheep 3374, 3376 and 3400 were injected
with the original citrated lamb's blood collected in the field. Another
 RZFT VALLE Y FEVER 553

sample of this lamb’s blood that had been collected in osalate-carbol-

glycerin mixture and stored in the ice-chest was inoculated into sheep
3370,3373 and 3375. Of the six sheep inoculated, three (3370,3374 and
3400) failed to react within seven days of the inoculation : the remainder
reacted typically after an incubation period of from 24 to 36 hours.

107. ~

a
CI
3

OVINE
104: 5
3361
c.

10.3: g /
2.

102:
.
p
c

.B
101.
1 2 3 I 5 6 7 8 9 10 11 12 l3 14 15 16 17 18.
DAYS AFTER MOCUIATION.

107:

OVINE OVIIl%

5568 s364.

DAYS mm NOCUWLTICN.

CHAW 111.
On the same day a fresh series of inoculations was also arranged
with stored citrated blood from the original ewe case. Sheep 3361,
3364 and 0369 received virus intravenously. All three reacted
(chart 111).
554 R. DAUBNEY AND J. R. HUDSON
Sheep 3361, a pregnant ewe, reacted on the morning of the second day, and its
temperature returned to normal on the third morning. On the eighth day, this
ewe aborted a lamb of approximately four months. Following the abortion there
was a sharp temperature reaction lasting four days, doubtless due to sepsis, and
the animal recovered.
Sheep 3364 reacted on the second day, and its temperature returned to normal
on the third morning ; there was a slight secondary rise on the fifth day, and the
animal was destroyed when very sick on the ninth day. At the time of death
there was a very profuse nasal discharge, and the animal, although in good
condition was too weak to stand. Post-mortem some lesions characteristic of the
disease were found, and histological examination of the liver confirmed the
existence of focal necrosis, although repair was commencing.
Sheep 3369 reacted on the second day and died on the fourth evening. On
post-mortem examination the following morning, decomposition mas advanced,
but the characteristic liver lesion was still evident. This was also a pregnant ewe
and she was lambing apparently a t or about full term, when death occurred.

I n view of the marked similarity between the reactions produced by

the two samples of blood, a cross-immunity test was arranged with
recovered animals. The results of this experiment are set out in
table I which demonstrates quite clearly the identity of the two strains
of virus.
TABLEI.
Cross-immunit?ytest between ewe and lamb viruses.
~~

original inoculation. lesult. Test inoculation. Rrsnlt.

3383 Lamb virus, 2nd passage + Lamb virus, natural case 0 (chart I)
3376 Lamb virus, natural case + ,, I, 0
3375 ,, (1 , + ,, 0
3380 Lamb virus, 1st passage + Ewe virus, natural case 0 (chart I)
3522 Lamb virus, 3rd passage + ,, ,, 0
+
9.

3548
3366
3379
,, ,, ,,
Ewe virus, natural case
Ewe virus, 3rd passage
; 1 9,

La;b virus, natural case

I,

91
0

:} (chart 11)
Lamb virus, natural case 1 +. Died 4th da:
Ewe virus, natural case

Looation of the virus in the tissues.

Blood-The early experiments had shown that blood taken during
the temperature reaction or at death contained the virus. With a view
to facilitating filtration the infectivity of the plasma was tested.
Two reacting sheep, 3439 and 3500, were bled into a solution of
0.9 per cent. NaC1. and 0.5 per cent. sodium citrate. The blood was
mixed and centrifuged, and when completely clear the citrated plasma
was decanted. The cells were washed twice in physiological saline
and resuspended. The plasma was inoculated into sheep 3440 and
3456, the corpuscles into sheep 3434 and 3453. All four animals
reacted typically.
 RIFT VALLEY FEVER 655

Licer. The liver is rich in virus. Minced liver tissue extracted

with a relatively small quantity of 9 per cent. NaCl and subsequently
diluted with a quantity of distilled water sufficient to reduce the salt
concentration to 0.9 per cent. furnishes virus suitable for inoculation
or filtration experiments, and has frequently been used for these
purposes.
4dee.n. The spleen also contains virus which may be liberated by
the same method of extraction.
Filtration.
A preliminary filtration experiment was carried out with lamb
virus of the second passage. Sheep 3383 was bled on 2nd August 1930
into citrate solution in distilled water. The hEmolysed blood was
diluted with eight parts of normal saline solution and filtered, using a
negative pressure of 630 mm. Hg., through a small british Berkefeld
filter. Sheep 3362 ancl 3363 were each inoculated subcutaneously with
5 C.C.of the filtrate. Both animals reacted typically and recovered.
A second attempt a t the filtration of the whole blood was made on
5th August 1930. Sheep 3522, a control animal reacting to unfiltered
blood from the previous experiment, was bled into citrate solution.
The citratecl blood was diluted 1 t o 20 with distilled water, and filtered
through large british Berkefeld candles and a small Seitz filter: the
diluted blood passed with difficulty through the Seitz. The Berkefeld
filtrate was inoculated into sheep 3349, 3354 and 3358, each of which
received the equivalent of 3 C.C. virulent blood. The Seitz filtrate was
iiiocnlated into sheep 3351 in a dose equivalent to 1.5 C.C.of the
virulent blood. The three animals inoculated with the Berkefeld
iiltrate all reacted typically after the usual incubalion period. The
sheep inoculated with the Seitz filtrate did not react, and was
subsequently proved to be susceptille.
JIi?acad hw. Livers from two natural cases, a ewe and lamb, were
received in fresh condition a t the laboratory on the 2nd August 1930.
%O grills. of niinced ewe’s liver were treated with 200 C.C. of 9 per
cent. sodium chloride solution ; two hours later distilled water was
added to make a total of 2000 c.c., and the liquicl was filtered through
Chamberland L 1 his, L 2 and L 3 candles. None of the animals that
received filtrate reacted. Control sheep 3448 and 3500, receiving
unfiltered extract, reacted on the evening of the third day.
An extract of the lamb’s liver was prepared in a similar manner.
This material was filtered through the Chamberland candles already
used for filtration of the ewe’s liver. Filtration was very slow, taking
seven hours. On the following day the L 3 filtrate was inoculated
into sheep 3502 and 3503, the L 2 filtrate into sheep 3496 and 3498,
the L 1 bisfiltrate into sheep 3436 and 3504. Sheep 3439 and 3452
received unfiltered material as controls. All the inoculations were
carried out intravenously.
556 R. DAUBNEY A N D J. R. HUDSON

Table I1 shows the results of the experiment.

TABLE11.
Filtration of extract prepared front the liver of a naturally
infected lamb.

Number of Filtrate.
sheep. Result.

3436 L 1 bis +. Recovered

3504 I, +. Destroyed 3rd day for examination ;
lesions typical
3496 L2 +. Died on 6th day ; lesions typical
3498 L2 +. Died on 6th day ; lesions typical
3602 L3 +. Killed in mttremis 7th day; lesions
typical of acute case
3503 L3 +. Recovered
Controls, Died 4th day ; lesions typical
unfiltered blood { +:0 Subsequently proved to be immune

It was obvious from the results of these experiments that filtration

of diluted blood or liver extract through the finer grades of Chamberland
candle was likely to prove unsatisfactory, because of the presence of
haernoglobin and cell detritus, Consequently, in later filtrations plasma
only was employed.
Filtration ofplasma. On the 28th August 1930, blood from a reacting
sheep 3508 was collected in an equal quantity of normal saline-citrate
solution. After separation of the plasma by centrifuging, it was
further diluted with eight parts of normal saline-citrate solution,
and filtered through Chamberland L 3, L 5 , and L 7 candles
at a negative pressure of 630 mm. Hg. Filtration was rapid, about
150 C.C. of the material passing through the L 3 and L 5 filters in
half an hour. The reaction of the material was tested before filtration,
and proved to be about pH '7.5.
The results of this experiment are given in table 111.
TABLE111.
Filtration of diluted plasma from sheep 3508.

Number of Filtrate. )ose : equivalent Result.

auimal. n C.C. of plasma.

I
Sheep 3681 +. Recovered
Lamb 3647 ] L3 +. Died on 20th day
Sheep 3682 +. Recovered
Lamb 3646 } L5 +. Died on 3rd day ; lesions
typical of acute form
Sheep 3677 +. Recovered.
Lamb 3638 } L7 +. Died in 48 hours : lesions
typical of acute form
.~ _ 1-
sheep 3678 _ .. Controls, . +. gec?vered- - .
 RlFT VALLEY FEVER 557

On 1st September 1930 and 5th September 1930 the filtration of

plasma was carried a stage further by the employment of the finest
grades of Chamberland candle. The conditions of the experiment were
identical with those detailed above. The essential data will be found
in table IV. Lambs, which we were now able t o obtain for experiment,
are more susceptible than sheep : of eight lambs inoculated with filtered
and unfiltered material only one survived.
TABLEIV.
Filtration of diluted plasma through the $ner grades of
Chamberland candle.

Nomber of Dosc : equivalent Result.

animal. in C.C. of plasma.
I I
Lamb 3627 . L7 1.75 C.C. +. Died within 48 hours ;
Lamb3629 . L9 1.75 ,. +.
lesions typical
Died 7th day; lesions
typical
,, +.
'1
i
Lamb 3631 1.75 Died 3rd day; lesions
Lamb 3649 L 11 typical
,, +.
Sheep3686
Sheep3697
1. L 13
1-75
4 C.C.
4 ,,
+.
0.
Recovered
Recovered
Later proved to be sus-
ceptible

I n view of the results already obtained with the finer grades of

Chamberland filter, a further carefully controlled filtration was under-
taken using L 2, L 9, L 11 and L 13 candles. Plasma from a reacting
lamb was diluted partly with normal saline, and partly with an eight-
day old culture of the organism of contagious bovine pleuro-pneumonia
in serum broth, so that the final dilution of plasma was 1 in 15. No
precaut,ions were taken to exclude contamination of the material before
filtration. The pH of the mixture was approximately '7.4. With a
negative pressure of 600 mm. Hg. filtration of 60 to 80 C.C.through
the L 2, L 11 and L 13 candles occupied about 3 minutes. Filtration
through the L 9 was slower, passage of the same quantity requiring
about 8 to 10 minutes. Five C.C. of each filtrate, equivalent to 2 C.C.
of filtered pleuro-pneumonia culture, were inoculated with the usual
precautions into a 100 C.C. bottle of serum broth. The L 2 candle
failed to give a sterile filtrate, and a rapid growth of contaminating
organisms masked any possible growth of the pleuro-pneumonia
organisin. The bottles inoculated with the I, 9, I, 11 and I, 13
filtrates remained sterile after a week's incubation. This result is in
accordance with the usual findings in filtration of the pleuro-pneumonia
organism, which has failed to pass L 5 candles in the experiments
carried out in this laboratory. Table V gives the results obtained
on inoculation of the filtrates into sheep.
It will be noticed that in this instance the virus succeeded in
passing an L 13 candle, although in a previous experiment with a
558 R. DAUBLVEY A N D /. R. HUDSON

TABLEV.
Controlled 3ltration of diluted plasma.

auimal.
Numberof I Filtrate.
I Dose : equivalent
in C.C. of plasma. Result.

.I
I
Lamb 3736 L2 I 1 C.C. +. Died 3rd day; lesions
typical
Sheep 3658
{ +. Recovered
Sheep 3662 } 2 C.C. +. Recovered
Sheep 3614
{ +. Recovered
Sheep 3666 } 2 C.C. +. Recovered
+. Recovered ; reaction
slightly delayed
+. Recovered

different candle of this grade it was held back. Similar irregular

results with candles of the finer grades have been recorded by Olitsky
and Boez (1927) with the virus of foot-and-mouth disease, which
passed L 7 candles once in 5 filtrations and L 9 candles once in
3 filtrations. It seems, therefore, that the virus of enzootic hepatitis
is in all probability smaller in size or is attached to smaller particles
than the virus of foot-and-mouth disease, since it regularly passes the
Chamberland filters up to the L 11grade.
I n all filtration experiments the inimunity of non-reacting animals
and of animals that reacted and recovered was tested by an inoculation
of unfiltered virus.
Other Properties of the Virus.
Minimal ilzfeetive dose. It has not yet been possible to determine
the actual minimal infective dose of blood. Observations on this
point should be carried out on young lambs, which unfortunately have
not been available in sufficiently large numbers for all our require-
ments. A single experiment has, however, been staged wibh adult
sheep, Blood from a reacting sheep was collected in citrate solution
and diluted with distilled water, the usual precautions being taken
to avoid any carry-over of virus. The dilutions were so arranged that
the dose to be inoculated was in each case 2 C.C. Sheep were inoculated
with doses corresponding to 0.04 c.c., 0.002 c.c., 0.001 c.c., 0.0005 C.C.
and 0.0002 C.C. of the whole blood. All the animals reacted typically,
and there was no lengthening of the incubation period in those that
received the smaller doses.
Xurvival of virus in vitro. When taken with precautions as to
sterility, the virus will remain viable in ordinary citrate solution a t
room temperature for at least a week. I n the cold room a t +_5"C.
the virus can be preserved in citrate or in oxalate-carbol-glycerin for
much longer periods. Tested after periods up t o 54 days, cold-stored
O.C.G. virus proved to be fully as virulent as fresh blood. After
 RIFT VALLEY FEVER 659

147 days in cold store the virus although still capable of provoking
reactions was undoubtedly considerably attenuated, the reactions being
poor and their appearance delayed. The cold-storage plant for seveyal
clays during this period was not functioning satisfactorily.
Precipitation of virus with protein fractions. The virus is pre-
cipitated from blood plasma with all the protein fractions obtained
by salting out with ammonium sulphate.
Certain experinients have been carried out in order to ascertain
the behaviour of different viruses when subjected t o salting-out by
electrolytes. As a precautionary measure precipitating solutions were
buffered a t about p H 7.4, to avoid any risk of destruction of virus as
the result of acidity. The protein fractions were each washed in the
precipitating concentration of the salt ; redissolved and reprecipitatecl
once or twice before the final solution was made. With the virus of
enzootic hepatitis, ammonium sulphate was the only salt employed
for precipitation, and each fraction was inoculated into two adult
sheep in closes corresponding to 0.25 C.C.and 2 C.C.of plasma. The
animals were inoculated with fibrinogen, euglobulin, pseudoglobulin
and albumin fractions ; all reacted, recovered and were subsequently
proved immune by test.
Animals Susceptible.
She cp.
The most susceptible animal is the newly-born lamb. Sheep of all
breeds and of all ages have proved susceptible in our experiments,
providing always that they were drawn from districts where the
disease was unknown. Mortality however is greatly influenced by
the age of the animal. The death-rate in newly-born lambs, oxperi-
mentally infected, is certainly in excess of 90 per cent. and it is quite
probable that this figure would hold for natural infection. Pregnant
ewes, experimentally infected, succumb in a proportion, including
deaths from complications following abortion, of about 20 per cent.
Of other classes of sheep utilised in our experiments, a definitely
lower mortality was experienced in weaners under eighteen months
of age. The experimental results are in agreement, so far as can be
ascertained, with field experience. It is a remarkable fact that a
filterable virus disease can regularly cause the death of lambs as
young as three days in the field, and that death can follow inoculation
of young lambs within 36 hours.
Goat.
The only goat inoculated with the virus gave a reaction, identical
with that shown by sheep, and recovered. NO outbreaks have been
reported in goats, although, when ewes have been aborting heavily in
an infected area, reports of abortions in pregnant goats have also been
receivect.
 560 R. DAUh'NEY A ND J. R. HUDSON

Cattle.
Cattle are susceptible to the virus, and a natural outbreak of the
disease has been observed in a dairy herd. This outbreak occurred on
a farm situated in Solai, in the Rift Valley, between 50 and 60 miles
from the first outbreak in sheep. There were no sheep on this farm;
but sheep and goats belonging to native squatters on a neighbouring
farm were reported to be aborting in large numbers. About 50 cases
of illness were observed in the dairy cows. There were three deaths
during the acute attack, and a further four deaths from sequuoln: such as
abortions. Symptoms were not very definite, attention being first
drawn to the illness by a more or less complete temporary cessation of
lactation. The animals' coats were staring and occasionally they
refused to feed. Salivation was a common symptom, and a t times
there was a f e t i d diarrhoea. There was never any laineness nor
stiffness of the limbs. I n most cases temperatures were not recordecl,
but during a visit to the farm, when three of the affected animals were
examined, one that was salivating and purging was found to have a
temperature of 107". Blood from this animal was despatched to the
laboratory for inoculation.
This blood was inoculatecl into two bullocks, two sheep known to
be immune to eiizootic hepatitis and two sheep susceptible t o the
disease. The bovines, which incidentally had been bred in the Rift
Valley, failed to react, as did the two immune sheep. The susceptible
sheep reacted typically, recovered, and subsequently proved to be
immune to a sheep strain of virus. These inoculations, together with
TABLEVI.
Comparison oj viruses of bovine and ovine origin.

Source of virus. Animal.

I Rrsult. Virus used for
immunity test. Result.

I
I:{
. 0
I ... ...

Cow, natural I Bullock

Immune 3357)
sheep (3687
Susceptible 3841
sheep
Cow . .
{
0
0
0
+.
Typical reaction
3642 +. Typical reaction
.
6509 + Died third evening;
typical liver lesion
...
) Sheep3683strain {
...
Immune
,,
,.
9,

,,
Her calf .5359 +. Typicalreaction
Passage of Cow. . 6127 +. Mild reaction
bovine virus Her calf . 5198 +. Typical reaction
Sheep strain
9,
Immune ; has been
hyper-irnmunised
through sheep
3642 Sheep , 3635 +. Destroyedfordiag- Immune
nosis ; lesions

Sheep strain
Bullock . 6039 +.
typical
Typical reaction,
chart IV
... ,.
Of ViNS Bullock . 6216 +. Atypical reaction; ...
died 17th day
 RIFT VALLEY FEVER 661

a further series of inoculations of the original virus, and a passage of

virus from one of the susceptible sheep are included in table VI.
Apart from dullness and inappetence the only symptom shown by
the experimentally infected cattle was the presence of a blood-stained
iuucous nasal discharge. The temperature reaction commences, on the
average, about 24 hours later than it does in sheep (chart IV).
Post-mortem the lesions resemble those produced in adult sheep.
106..

105'.

6039.
102:
IrnUnS t o
Pl"ZeF*e,t.

101..

100:

C~IART
IV.

Jh.
During the course of the investigation all the four Europeans
engaged developed a dengue-like fever, which we now know to have
been due to infection with this virus. The first person to be attacked
was Mr F. Lyons, the laboratory assistant in charge of the exposure
experiment on the farm where the disease first appeared. Mr E. J.
Hull, w h o assisted i n the laboratory investigation, was next attacked.
A few days later, both writers developed symptoms within a few hours
of each other.
- I n every case the onset of the attack was characterised by a very
brief period of general malaise, followed rapidly by sharp rigors and
headache. The temperature rose t o about 103", and the face was
brightly flushed. I n from 3 to 6 hours after the rigors had passed
off, pains developed in or iiear the joints extending from the
base of the skull to the extremities. Fever persisted for a period of
from 1 2 to 36 hours, and the pains gradually disappeared within about
four days. One of us, Mr Hall, had a second temperature reaction,
accompanied by similar symptoms, three days after recovery from the
initial attack : he complained of headache and defective vision for
son18 weeks afterwards.
JOURN. OF PATH.-VOL. XXXIV. 2P
 562 R. D A UBNE Y AND J. R. HUDSON

Upon enquiry we found that almost every native engaged in herding

sheep during the original outbreak had been ill for some four days and
hacl complained of fever accompanied by severe pains. At the time
these illnesses were ascribed by the manager to eating the somewhat
decomposed carcases of dead sheep.
The manager of the farm in Solai where the outbreak in bovines
was discovered, also contracted the disease; in fact it was his description
of what he considered to be an attack of influenza that made us suspect
the true,nature of the disease occurring in his cattle. He hacl been a
chronic malarial subject for some years before the attack of Rift Valley
fever, and stated that after recovery he experienced no further malarial
relapses. It seemed possible that the stimulating effect of the virus
disease upon the reticulo-endothelial system (as shown by the hyper-
trophy of the endothelial nuclei of the liver sinusoids) niight have
enabled the patient to rid himself of his malarial parasites. Since
upwards of two hundred human cases of the disease were known to
have occurred without any fatality experimental inoculation appeared
t o involve no risk, and it was decided to inoculate a patient sugering
from subtertian malaria a t the Native Hospital, Nairobi, with the
virus, in order t o observe its effect on a malarial infection.
The man was an adult of the Kisii tribe, admitted to hospital on
24th December 1930, when suffering from a relapse of subtertian malaria.
The attack was mild, it was not treated with quinine, and the parasites
disappeared from the circulation on 5th January. I n order to ascertain whether
recovery was complete or not, one grain of tartar emetic was given intravenously
on 13th January, and a recrudescence of nialaria ensued. Parasites persisted
in scanty numbers for several weeks after the provoked attack.
At 2 P.M. on 20th January, 3 C.C. of diluted filtered virus (see table VII)
TABLEVII.
In o cu l atims with plasma f?.on& lamb 3802 o n 20/1/1931.

Sumtier. XIater!al and equivalent dose of Remarks.

-
~~-

Native 4Oi6 0.2 C.C. of Chamberland L 6 filtrate ......

Sheep 3784 Typical reaction. Immune
when tested subsequently.
Reaction confirmed by sub-
inoculation into susceptible
and imniune sheep.

were inoculated intramuscularly into the buttock. Daily observations were

made on the blood and urine. -At 4 P.M. on 23rd January; the patient became
ill with headache and pains in the loins, but no febrile reaction took place
until the following day, when the temperature (axillary) rose to 103'4°F. and
symptoms increased in severity, pain being particularly severe in the bones of
the legs and in the left hypochondrium. The face was congested, swollen and
moist, the eyes were slightly bloodshot, the pulse rapid and bounding. There
was no vomiting and the tongue was only slightly furred. There was no
 RIFT V A L L E Y FEVER 663

palpable enlargement of the liver or spleen. The patient felt very ill, remaining
in one posture in bed, and not tossing about as in a malarial paroxysm. In
the evening, much generalised pain was felt in the abdomen, although there
were no tender spots or rigidity. The temperature returned to normal on the
evening of the next day, and the patient felt much better except for pains in
the bones of the legs andof the ankles. Abdominal discomfort remained and
persisted on and off for weeks; in fact on the 23rd of February, the patient
was still complaining of pains in the abdomen and diarrhoea, although the
latter did not make its appearance until the middle of February. Apparently,
rrcoverv from the attack of Rift Valley fever was rapid, but the clinical
picture was somewhat obscured
by a further recrudescence of
inalnria on the 28th of January,
which became so severe that
for the first time in the malarial
history of the case quinine had
to be administered, and the
subtertian infection dispersed
by this agent.
The secondary rises of tem-
perature must be considered as
malarial in origin, since daily
examination of blood slides
showed an increasing number
of subtertian rings, leading up I
to the paroxysms of the 28th
and 29th of January, which
incidentally were accompanied
by symptoms much more sug-
gestive of malaria than of Rift
Valley fever. The patient was
discharged from hospital on
4th February, well except for
pains in the ligaments and
muscles of the back, near the
left scapula.
The leucocyte findings are
shown together with the tem-
perature on chart V. The
leucocyte counts were per-
formed daily a t 9.30 A.M., about
t h e e hours after a meal. On
the dav the svmntoms first
" 1

appeared (23/1/31), a polymorphonuclear leucocytosis of 5000 per c.mm. occurred,

which was followed by a drop to nearly half on the next day, and by a further
drop to 1100 per c.mm. on 27th January. The counts after this date follow
the sequence common to malarial relapses, a leucocytosis during the paroxysrri
followed by a leucopenia. The patienti count gradually returned to normal.
The polyrnorphonuclear-lymphocyte ratio throughout more nearly approximated
to unity than in normal people, a condition, however, that is commonly
encountered among native races in Kenya. Metamyelocytes, band-forms and
one-lobed polymorphonuclear leucocytes were frequently seen in the blood-
smears, equally in December and during the attack of Rift Valley fever. This
also is a common feature of native blood, and no increase in their number or
greater shift to the left in the Arneth index was observed during the illness.
 564 R. DAUBNEY AND J R. HUDSON
There were no significant changes in the red blood corpuscles, the eosinophils
or the large mononuclear cells.
The van den Bergh reaction of the blood-serum was negative throughout.
The urine showed a faint trace of urobilinogen before and during the attack,
but the presence of albumin was not detected until 25th January, from which
date it persisted in varying amounts (always scanty), until 16th February, when
tests were discontinued.
Irwcdationsfrom the patient. Blood was taken from the patient daily from
the 24th January (the day on which the temperature rose) to the 4th February,
and a small quantity of each sample was inoculated into susceptible sheep to
test the infectivity of the blood. From the 24th to the 29th of January the
daily samples proved to be infective (table VIII), but from then onwards to
the 4th of February only negative results were obtained.
TABLEVIII.
Inoculations from human case of Rqt Valley fever during the reaction.

Day
on which Iiumber
material Inooulum. of animals Result of inoculation. Subinoculation. Itiimui~itytest.
was inoculated.
collected.

24/1/31 2 C.C. serun Lamb 3803 Temperature reaction 27/1/31 into lamb 9/2/31, immune
only 3805, reacted,
died, lesions

25/1/31
‘2c.c. (. ,, 3807 Temperature reaction
only
typical
... 9/2/31 ,,
15 C.C. urine ., 3515 No reaction ..I 5/2/31 reacted and
died
28/1/31
?2C.C. serun Sheep 3774 Temperature reaction
only
... 9/2/31, immune
.5 C.C. urine Lamb 3813 No reaction ’ ... 5/2/31, reacted
severely
27/1/31 2 C.C. serun Sheep 3777 Temperature reaction ... 9/2/31, immune
only
28/1/31 ,, ,, 3i94 Delayed temperature
reaction
... 9/2/31 ..
29/1/31 ,, ,, 3780 Delayed temperature ..I 9/2/31 ,.
reaction
30/1/31 ,, 3775 No reaction ... 9/2/31, susceptible
31/1/31 ,, 3776 9/2/31 ,,
I, ,t

...
. . I

1/2/31 ... ...... ...

2/2/31 Sheep’3797 No reaction ..I 9/2/31, susceptible
3/2/31 Lamb 3821 ,, 1. ..I 9/2/31 ,,
4/2/31 , ,, 3825 7. ,, . . I 2/2/31 ,,

Urine collected on the 25th and 26th of January failed to infect susceptible
lambs, and no further tests were made.
There was some indication that the virus recovered from the patient, even
at the height of the temperature reaction, was not so virulent for lambs as
usual; but that a single passage through lambs was sufficient to restore its
virulence completely. For example, lamb 3803, inoculated with the blood-
sample of the 24th January, showed a brief temperature reaction only, while a
second lamb, inoculated from 3803 during the reaction, died on the third day
after inoculation in the usual way.
A small quantity of the patient’s serum collected two weeks after recovery
was tested for protective power. Two lambs, 3859 and 3858, twins of the same
sex, were inoculated each with 3 C.C. of serum, the former receiving a normal
human serum and the latter serum from the patient. Both received in addition
 RZFT VALLE Y FEVER 565

an injection of 1 C.C. of virulent blood. The lamb 3859, treated with normal
serum, was dead by the forty-eighth hour, whereas the other lamb showed a
delayed temperature reaction only and never appeared to be in the least unwell.
It is interesting to compare the clinical and laboratory findings in this case
with the known pathology of the disease in sheep and cattle. The incubation
period was 74 hours, about the same as in cattle and slightly longer than
in sheep. There was no palpable enlargement of the liver, and no disturbance
of liver function could be detected by van den Bergh’s test or by examination
of the urine. The presence of albumin and the absence of casts suggests the
existence of a nephrosis, a constant feature of the disease in sheep. The
abdominal pains are highly suggestive of the characteristic mesenteric lesions
seen post-morteh in lambs that have died from enzootic heapatitis, although no
actual enlargement could be made out. The blood changes resemble those found
in sheep, but were not sufficiently severe to cause a n outpouring of the more
primitive elements of the haemopoetic system.
Diagnosis of the disease must always be a matter of some difticulty, although
in known enzootic areas the presence of a short febrile illness, characterised by
prostration, pains in the bones, ligaments and joints, abdominal discomfort,
absence of enlargement of the spleen, and a leucocytosis followed by a progressive
leucopenia should be sufficient to establish diagnosis.
Malaria is easily differentiated by the nature of the paroxysms and the
presence of parasites Dongue fever may be excluded in the absencc of rash,
of enlargement of glands and of the characteristic saddleback temperature chart.
The leucocytic changes in the two diseases are very similar, even to the progressive
leucopenia which reaches its lowest point after the termination of the pyrexia
(Simmons, S t John and Reynolds 1931). Influenza presents more difficulty
although there is a leucocytosis throughout in this fever, and the definite localisa-
tion of pains in the bones and joints in the one disease and the frequency of
catarrhal symptoms in the other are all of assistance in separating the two
conditions. Sandfly fever, three-day fes7er and other fevers whose &iology is
more obscure resemble Rift Valley fever in many ways; in fact it is not
impossible that some outbreaks of Rift Valley fever have already been referred
to this group. There is, however, a certain method of differentiation, i.e. by
inoculation of lambs with blood taken during or soon after the febrile period.
So far as the effect of the disease upon a chronic malarial infection is concerned
it may be stated that in this instance no evidence of increased resistance to the
malarial infection was obtained, on the contrary the evidence pointed rather t o an
immediate aggravation of the malaria. The fact that a further parasitic relapse
was observed on the 23rd of February, a month after the attack of Rift Valley
fever showed that no development of immunity occurred within a reasonable
period after the attack.
Hovsc.
Only one horse has been inoculated with virus, and this nniiiial
failed to react.
Pig.
Two pigs have been inoculated with virus taken from nil acute
case in a lamb; neither of the two reacted in ally may.
Natural transmission.
From the earliest experiments, susceptible sheep have been housed
with those reacting to virus. In spite of the fact that the cxperi-
mentally-produced disease differs in no detail from the natural disease
JOURN. OF PATH.-VOL. SXXIV. 2 r 2
566 R. DAUBNEY AND J. R. HUDSON

iufection by contact has never occurred. When subsequently

tested on their immunity contact animals have invariably proved
susceptible.
Experiments in the field designed to test the possibility of trans-
mission from ewe to lamb by contact and by feeding milk broke down
owing to the rapidity with which the disease spread to all parts of the
farm, including the buildings. At the laboratory, nursing ewes have
011 occasion been inoculated with virus, have reacted, and have even
died without infecting their lambs. Conversely, young lambs have
frequently been infected by inoculation and have died without infecting
their mothers. Lambs have also been fostered on ewes whose own
lambs have been killed by the experimental disease without any
transmission taking place.
To sum LIP, one can state that a disease which in the field was
able to assume the proportions of a serious epizootic, failed to spread
naturally in the sheep population a t the laboratory, in spite of the
close contact maintained and of the proved susceptibility of the great
iiiajority of the animals. The most feasible explanation of this state
of affairs would be that the disease was indirectly transmitted from
animal to animal and that the necessary transmitting agent was absent
from Rabete, where the laboratory experiments were carried out.
Further light is thrown on this problem by the result of a field
exposure experiment that we were able to arrange on the farm where
the disease first occurred.

Nineteen sheep were divided into three batches and exposed under different
conditions. Six sheep were placed in cages covered by ordinary mosquito nets,
and were kept under cover all the time. Five sheep were kept in similar
iliosquito-proof cages at night and were allowed to graze with the third batch
during the day : these five sheep were housed every evening before sunset and
were not let out until after seven o’clock in the morning. The remaining eight sheep
grazed during the day, and a t night were enclosed in a small ring fence composed
partly of the cages containing the other sheep and partly of branches of Acacia.
Good experimental conditions were maintained for a period of five weeks,
during which time a number of the animals showed temperature reactions, and
one died of enzootic hepatitis. Whenever one of the experimental animals
showed anything that might be interpreted as a reaction-even the slightest
rise in temperature-it was bled, and the blood was despatched to the laboratory
for subinoculation into susceptible sheep. The immunity of sheep used for
subinocu1,ztions was later tested by inoculation with virus. At the conclusion
of the field experiment the exposed sheep were returned to the laboratory,
where they mere subjected to a controlled immunity test with virus. Table I X
shows the result of this exposure together with the subsequent immunity test
of the exposed animals.
It will be observed that of the six animals kept all the time under mosquito
nets, every one was still susceptible when tested at the conclusion of the
experiment j and that although blood was taken from three of these animals at
the time of slight temperature reactions, transmission to susceptible sheep was
never effected.
Of the five animals protected at night one showed a reaction during the
 TABLEIX.
Field exposure experiment.
Result of final
Rosult of subinoculation, and of subsequent rnmunity tost of origina:
Number of sheep. Date of reaction in the field. Subinoculation. immunity test of iiioculated animal. exposed sheep.

2654 3/9/30. Died 6/9/30 . . . 6/9/30. Sheep 3638 Died 7th day. Lesions typical ...
3493 6 to 8/8/30 . . . . . 16/8/30. ,, 3444 Died 4th day. Lesions typical Immune
3710 2 to 4/9/30 . . . . . 6/9/30. ,, 3639 Died 6th day. Lesions typical ,,
21/8/30. ,, 3442 Slight reaction. Immune to Test 368( .,
Out all 3429 . . . .
14/8/80 ,, 3447 ,, , 9. ,,
the time 3315 Doubtful 16/8/30, 104'4"
4
. . 6/jj30. ,, 3696 No reaction.' Susceptible at Test
1~ Susceptible
3711 4/9/30, 104-4" . . . ,, 3679 9,
Immune
3713 29/;)3G . . . . . . N($ bled ... ,,
3712 No reaction . . . . . ... ... Susceptible

3382 Doubtful reaction 1/9/30 . . Sheep 3633 No reaction. Susceptible Susceptible

3426 ,, 15/9/30, 103.2" . ,. 3630 ,, 3. ,.
In all 3511 ,, 5/9/30, 104.4" . ,, 3691 ,9 9 ,,
the time 3717 No reaction . . . . . ... ...... ,,
3718 ... ...... 99
I, . . . . .
3716 ,9 . . . . . ... ...... ,,

3325 Noreaction . . . . . ... ...... Susceptible

3714 . . . . . ... ...... 1,
In at ,*
night 3715 . . . . . ...... Tmmunc
..
3366 1Iv:iction 28/8/30, 105" . . . Noi'ilrd ......
Xj6i No reactioil . . . . . ... ...... Siisctsptible
568 R. DAUBNEY AND J R. HUDSON

fourth week; but through an oversight the blood of this sheep was not
subinoculated. At the immunity test, this animal alone out of the second
batch proved to be immune to enzootic hepatitis.
Of the eight animals exposed day and night, seven showed some form of
temperature reaction during the period of exposure; one of them actually died
of enzootic hepatitis.
Positive results were obtained with four out of six of these bloods when
subinoculations were made a t the laboratory. The remaining reacting animal
was not bled at the time of its reaction, but subsequently proved to be
immune. Of the seven survivors of this batch tested a t the close of the
experiment, only two were still susceptible, one in which no reaction had been
recorded during the exposure, and onc of the two doubtful reactors that had
failed to transmit by inoculation. There is a very satisfactory agreement
between the results of subinoculation of blood from the reacting animals and
the findings in the final immunity test.

On the strength of this experiment it was concluded that in all

probability transmission is effected by some insect that is sufficiently
large to be excluded by an ordinary mosquito net and which feeds
chiefly a t night. This hypothesis is supported by certain features in
the history of the original outbreak. It will be recalled that on the
10th August the remaining lambs and ewes were moved to a higher
altitude, and that in spite of the occurrence of deaths on the road
and for a few days after arrival, there was every indication that the
manceuvre had been entirely successful in checking the outbreak. It
is logical to conclude that some vector that had been operating at the
lower level was absent a t the higher. As a matter of fact mosquitos,
which have been particularly numerous in the valley this year, are
rarely seen on the higher pastures.
Systematic catching of mosquitos has been carried out on the
lower farm since late September. Catches have been determined and
sorted by Mr Roberts, Senior Entomologist, a t the Medical Research
Laboratory, Nairobi, t o whom we are indebted for much valuable
assistance in connection with this aspect of the problem. The great
majority of mosquitos taken proved to be either Culex pipiens or C.
fatigum. Three other species, Anopheles spuarnosws, A. mauritianus and
Ta?niorlLynchus brevipalpis, have been collected in small numbers.
Many hundreds of the two species of Culex have been inoculated
i n t o susceptible sheep, and anything that might be interpreted as a
reaction has been investigated by subinoculation and immunity test.
So far no evidence of transmission has been obtained.
A similar procedure has been followed with the few available
examples of the other species ; in the case of the Anophelines equally
without success.
One lamb inoculated with an emulsion of five Tceniorhynchws
brevi-palpis, three of which were rather decomposed, showed no
temperature reaction, but was subsequently found to be immune.
Fortunately blood slides were taken during the first few days after
 FIG.I.--Iiidney : naturd C L S ~: ewe. Frc. 2.-Liver : experimental ciisc : sheep.
H. anti R. H. and E.

FIG.:<.-Liver : experimental case : sheep. F I I ; 4.-Liver

. : n;tturiil case: lamb.
H. rind E. H. tinct E.

14’1~. 5.-Liver : intr:inuclcar bodies : csperi- FIG.6.-Liver : liyaline bodies : ixttural

nieiit;il case : sheep. Goodpastiire’s c:ise : cwt:. Mtinn’s stain.
stain .
 RZFT VALLEY FEVER 569

inoculation, although they were not examined till two months later.
The smears taken on the second third and fourth days show the
progressive leucocyte changes which we have come to associate with
this disease. This lamb was born at the laboratory and could not have
acquired immunity except from the inoculation of Tcmioyhynchus.
This evidence points strongly to Tmuiodynchus as the vector, and it
is intended to continue systematic catching throughout one coiiiplete
year at least.
Histology.

Spleen.
Histologically, there is little to remark in the spleen. The sinusoicls
are swollen; but do not contain an excessive number of white cells.
The subcapsular haemorrhages are not extensive. I n an occasional
case there is some evidence of nuclear degeneration, affecting a few
cells in the pulp. When seen, these nuclear changes consist of a
condensation of the nucleus, sometimes followed by karyorrhexis.

Kidney.
Essentially, the pathological change in the kidney is a tubular
degeneration or nephrosis. In hyperacuta cases in lambs, either
naturally or experimentally infected, death takes place too rapidly
to permit the development of any extensive lesion. I n such animals
there is cloudy swelling of the epithelium of the convoluted tubules,
some slight congestion of the vessels both of the cortex and the
niedulla; but no increase in the number of polymorphonuclear
leucocytes present in the tissues. Occasionally in lambs the lesion
may have progressed a stage further towards a definite tubular
degeneration. The epithelial nuclei, although still taking the stain
may be noticeably pale, the cytoplasm of the cells granular, and their
free edges frayed.
Adult sheep may live long enough for the development of a
definite tubular nephrosis, and the actual lesion found post-mortem
depends on the length of time that they survive. The change begins
in the convoluted tubules with loss of nuclear staining and condensa-
tion of the cytoplasm of the epithelium. I n the more acute cases
the change in the convoluted tubules may be quite advanced, while
the remaining tubules and the glomeruli appear normal. I n such
kidneys, the only other change appears to be a slight congestion of
the medullary and cortical vessels. The kidney of sheep 3439 (chart
VI) which died four days after inoculation illustrates this type of
cortical lesion and furnishes some indication of the rapidity with
which the lesion develops (fig. 1). I n the less acute cases the
tubular change is less advanced ; but may be found affecting equally
all the tubalar epithelium of the cortex, and to a less extent that
570 R. D A U R N E Y A N D /. R. HUDSON

of the medulla. The glomeruli do not contain an excessive number

of nuclei and Bowman’s capsule remains unaffected.
If sheep survive seven days it is usual for them t o recover. About
the ninth day one finds, in addition to the tubular changes described
above, that the glomeruli contain more nuclei than normal. The
excess consists chiefly of polymorphonuclear leucocytes. Some of the
~ nuclei,in all probability polymorphonuclears,
have undergone rhexis. It is the esception
to find any proliferation of the cells of
Bowman’s capsule. Congestion of the
vessels of the cortex and niedulla is still
maintained, and the connective tissue nuclei
appear swollen and stain deeply; in the
medulla there may be evidence of prolifera-
OVINE
tion.
3439 The process of repair is remarkably slow,
in striking contrast t o the rapidity with
which fibrosis is effected, following the
tubular nephritis of Nairobi sheep disease.
I n one animal, killed two months after
infection, the lining of the majority of the
cortical tubules still remained incomplete
D l y S AFTER MOCUIATIOA
and there was little evidence of epithelial
CHART VI.
repair. While the nuclei of the fibroblasts
of the cortex appeared to be swollen aml
deeply-stained, there was no tendency towards deposition of fibrous
tissue.
It is evident that the kidney lesion is merely a toxic degenerative
change and like similar lesions in other acute fevers, e.g., yellow fever,
it is not followed by fibrosis.
Lung.
Frequently the lung shows nothing abnormal, but in slioep that
have been unable to rise for some time prior to death there may be
a pronounced unilateral or bilateral hypostatic congestion. The small
white raised areas on the surface of the lung, to which referelice has
already been made, prove to be due to the distension of the alveolar
capillaries with leucocytes, most of which are polymorphonuclears.

Cimdaiory systeni.
There is nothing characteristic about the hzmorrhages iu the heart
muscle.
Blood.
At the commencement of the reaction there is a very marked
increase in the proportion of polymorphonuclear leucocytes, and at
the same time in young lambs there is an absolute decrease in the
 RZFT VALLEY FEVER 571

number of monocytes. The number of polymorphonuclear leucocytes

soon begins to fall sharply; in lambs the fall is noticeable some
24 hours after the reaction commences, and young forms comprising
some 50 per cent. of myelocytes and metamyelocytes appear. At
the time of death normal polymorphonuclear leucocytes arc
usually absent from the peripheral circulation and monocytes are
rarely seen. The white cells of the blood consist almost entirely
of myeloblasts, large numbers of which show Auer granules, and
lymphocytes. I n young lambs this sequence of changes usually
occupies three days only.
No systematic daily examination of the blood of adult sheep has
been carried out; but smears taken a t the time of death from both
natural and experimental cases give the same blood picture as the
slicles taken a t death from young lambs.
No changes have been remarked in the red cells, and no
micro-organisms or unusual bodies have been detected in blood
s inears.

Lymphatic glands.
Those niesenteric glands which macroscopically are enlarged and
show hemorrhages in the cortex reveal the expected lesions on histo-
logical examination. Hmnorrhages from the larger vessels occur in the
neighbourhood of the cortical lymph nodules, usually more centrally
placed than the nodules theinselves. When the hsmorrhages arc
extensive and accoinpanied by polyniorph infiltration it is common
to find inany of the leucocytes undergoing karyorrhexis both within
the nodules and in the fixed tissue of the gland. Distinct necrotic
foci have never been found in lymphoid tissues.

Alimentary truct.
The inost noticeable lesion of the alimentary tract is the catarrhal
enteritis already mentioned as occurring in the m a l l intestine. The
lesion is characterised by clesquamation of the epithelium and erosion
of the villi. The interglandular tissue is infiltrated throughout by
large numbers of polymorphonuclear leucocytes which are particularly
numerous in and around the lyniphoicl follicles. Karyorrhexis of the
iiuclei of the infiltrating cells is a striking feature in the lyinpli
nodules.
It will be recalled that reference has been made to the occasional
occurrence in naturally infected adult sheep of a hemorrhagic or
(ILarry ” enteritis. The “tarry” mass filling the lumen of the gut in
these cases consists largely of coagulated blood in which is embedded
epithelial detritus, leucocytes and bacteria ; the nuclei of both epithelial
cells aiicl leucocg tes are in various stages of dissolution.
572 R. DAUBNEY A N D 1.R. N U B S O N

Nwvous system.
Apart from injection of the meningeal vessels, which is usually
discernible to the naked eye, no lesions have been found in the
central nervous system.
Livs?..
Histologically the liver lesion is in every way as diagnostic as the
naked eye appearance of the organ (figs. 2 to 6). One might go
further and state that whenever the macroscopic changes are
insufficiently pronounced t o enable a diagnosis to be made, which is
not infrequently tlie case in adult sheep, histological exaniination will
enable one to establish the diagnosis beyond any possibility of error.
Although the general picture may vary with the susceptibility of the
subject and with the degree of rapidity with which a fatal termination
or recovery is reached, the nature of the process remains essentially
the same in all cases. It may be characterised briefly as a focal
degeneration of liver cells closely followed by infiltration of the lesion
with phagocytic cells, chiefly polyniorphonuclear leucocytes and
histiocytes. The general sequence of these changes can most easily
be followed in sections from adult sheep, since in young lambs the
whole process is so rapid and the destruction of tissue is so enormous
as to obscure completely the actual nature of the lesion.
In hyperacute cases in young lanibs a section of the organ niay be
unrecognisable as liver. Usually, however, a few hepatic cells remain
a t some portion of the periphery of the lobule, and in an occasional
lobule a few liver cells may be recognisable in the immediate vicinity
of the central vein. For the rest, the lobule is completely filled by
infiltrating cells, the faintly-staining cytoplasm of degenerated liver
cells, and aiiiorphous granular masses of chromatin in which here and
there one can find the more or less degenerated nucleus oE an hepatic
cell. The infiltrating cells are for the most part polymorphonuclear
leucocytes, with a certain number of wandering histiocytes. The
former display a very noticeable tendency to undergo karyorrhexis,
and the masses of chromatin are derived in part from them, and in
part from the disintegrated nuclei of hepatic cells. All that remains
of the cytoplasm of the liver cells in these areas is a number of round
or oval fragments of homogeneous material, each representing a liver
cell that has eviclently undergone the characteristic hyaline degeneration
that is described below. They have, a t this stage, however, almost
completely lost their affinity for eosin.
Where sections of liver are available from adult sheep at varying
periods after infection, it is possible to follow the development of the
lesion step by step. The earliest change inay affect either a single
hepatic cell, or a group of from two to five cells, situated anywhere in
the lobule ; but more frequently in the central zone. The affected cells
 RIFT VALLEY FEVER 573

stand out clearly on account of the increased affinity for eosin of their
cytoplasm, which is homogeneous, and apparently denser than norma1 :
a t this stage there may be no evidence of nuclear change. A more
advanced lesion will involve a greater number of cells, up to two-thirds
or more of the entire lobule. I n the later lesion the cells have under-
gone a certain amount of contraction, have become rounded or oval,
and are more or less completely separated from one another: and
nuclear degeneration is now apparent.
The next step i n the process is an infiltration of the focus by
polyniorphonuclear leucocytes and histiocytes, chiefly the former.
Large numbers of lhese infiltrating cells soon degenerate, and one
finds masses of granular chromatin derived from the nuclei of both
infiltrating and paraenchymatous cells. Generally when the lesion
begins in the central zone, the central vein is occluded by the necrotic
mass and little or no trace of its wall remains. OnIy in very few
cases is there evidence of distension of the central vein with blood and
hmiorrhage or diapedesis in to the lobule.
I n the vicinity of the foci the endothelium of the sinusoids
commonly appears Rwollen, and the nuclei stain deeply. I n a
proport,ion of cases the sinusoids throughout the affected lobules,
and this may mean throughout the organ, are clistended with eryth-
rocytes; as a rule, however, they show no excess of blood. When seen
this congestion of the sinusoids occurs most frequently during the
second week of the reaction in adult sheep that are progressing towards
recovery.

It has been possible to study the process of repair in sections of

liver from recovered animals killed a t varying periods after the
reaction. By about the tenth day the foci are not so marked; and
although they niay contain some chromatin fragments and clebris of
hepatic cells, the infiltrating polymorphonuclear leucocytes are much
less nnnierous and are found chiefly at the periphery of the lesion.
The sinusoids throughout the organ are dilated, and the liver cords
appear to be widely separated. In a few cases to which reference has
already been made, the sinusoids are packed with erythocytes ; but more
generally they appear empty except for a few polyinorphonnclear
leucocytes and small round cells. The endothelial nuclei are swollen,
rounded, and stain intensely, and there has undoubtedly been some
prolifenLtion. Traces of the foci remain, in some cases two months
after the reaction; but a t that stage they contain few or no chroniatiii
fragments, and there are only a very few infiltrating polymorphonuclear
cells at the margin of the lesion. The sinusoids are still dilated and
the endothelium continues to be active. By two and a half months
recovery may be complete, and the liver may show no trace of injury.
Cirrhosis has never been observed as a sequel to the destruction of
hepatic tissue.
674 R. DAUBNEY A N D 1.R, HUDSON
Sections from but two bovines have been available for histological
study. One of these, an acute experimental case, shows a focal necrotic
lesion of the liver similar to that described in sheep. The material
from this animal was unfortunately too deconiposed to allow of any
accurate study of the nuclear changes ; but the general features of the
foci were those of hyaline degeneration followecl by infiltration. The
second case survived seventeen clays, and a t death the liver contained
no focal lesion.

Cytology of the cell changes in the liver.

From the outset it was realised that the cytoplasmic changes in
the hepatic cells (figs. 5, 6, 8 to 14) bore a striking reseniblaiice to
what is generally known as the “ Councilman lesion in yellow fever
))

of man. Unfortunately, Councilman’s original conirriunicatioii was not

available, and the description given recently by Torres (1928) was much
contracted While this manuscript was in course of preparation,
Rlotz and Belt (1930) reclescribed the Councilman lesion in ternis
that only served to emphasise the resemblance between the two
morbid processes.
The earliest change in the cell is cloudy swelling, which js rapidly
followecl by hyaline degeneration of either a part or the whole of the
cytoplasm (fig. 8). The hyaline mass is at first irregular in outline;
but soon becornes rounded off into a sharply-defined oval or spherical
niass. The rounding-off process involves some contraction of the mass,
and where the entire cell is iniplicated this results in complete
separation of the individual froin its neighbours. When only a part
of the cell is affected the contraction separates the hyaline body from
the healthy cytoplasm, ancl the mass is surrounded by a iiiore or less
distinct, clear halo. In large foci, such as occur in the more acute
cases, the degeneration almost invariably affects wliole cells, ancl liyaline
bodies are extremely rare. Wheu found, they are usually a t the
periphery of the focus. In a less acute case, where the foci are small,
hyaline bodies may be the most striking feature of the section, occurring
free in the foci and at the same time in the cytoplasm of otherwise
healthy cells throughout the organ. It is quite evident that the
hyaline bodies are frequently extruded from cells in the niwe iiornial
parts of the tissue.
The hyaline bodies (figs. 6, 14) resemble those of yellow fever in
size and in staining reaction ; the vacuolatecl appearance described by
Klotz and Belt (206. cit.) as due to included fat is not coniniou: fatty
changes are indeed exceptional in this disease. The bodies are best
denionstrated by such stains as Mann’s, Borrel’s magenta-picro-indigo-
carmine, and Goodpasture’s aniline-fuchsine. When stained by Mann’s
method they resemble in striking fashion the Negri bodies of rabies,
and it is difficult t o point to anything in their structure or Inode of
JOUILNAL OF PATBOWQY.-VOL. X X X I V . PLATEXLTI

E N Z ~HEPATITIS
C (RIIT VALLEYFEVER)

”
Q
Q)
 14
Fro. 8. -Liver : experimental case : sheep. Goodpasture’s stain.
FIGS.9-13.-Liver : intranuclear bodies : natural
case : ewe. Mann’s stain.
FIG. 14.-Liver : hyaline bodies : natural case :
ewe. Mann’s stain.
 RIFT Vd LLE Y FE V E X 576

development that can serve to differentiate them from inclusion bodies.

Any distinction must rest upon the fact that the process of hyaline
degeneration can and frequently does involve the whole of the cytoplasiii
of the cell. Klotz and Belt have remarked upon the reseniblaiice
of the hyaline bodies of yellow fever to the cytoplasmic inclusions
in other virus diseases and in particular to Negri bodies. Although
there is n o intention to suggest that they are specific inclusion
I)oclies in the generally accepted sense, me feel with them that there
is a very definite analogy. I n the later stages the hyaline cytoplasiii
becomes iiiuch less markedly acidophilic, and in the centre of an old
lesion the cells may be quite pale, although their homogeneous
appearance remains.
The nuclear changes in affected cells are of the type generally
lmown as oxychroniatic degeneration (Landa and Luger, 1926).
Nuclear changes may be initiated in advance of any cytoplasmic
cliange other than a cloudy swelling, or they may not make their
appearance until hyaline degeneration of the cytoplasm is well-
advanced. One frequently finds intensely aciclophilic nuclei in cells
tlistribu tecl throughout the more normal parts of the lobule and show-ing
little or no cytoplasniic change. It is unlikely that these acidophilic
nuclei have any relation to the sequence of nuclear changes that take
place in the degenerating cells of the foci. I n the latter, the nucleus
first beconies hyperchromatic ; then the basophilic elements tend to
be distributed towards the periphery, and amorphous or granular
acidophilic material appears in the central area. The nucleolus becomes
inore strongly basophilic, and may move to the margin. Hypertrophy
of the nucleolus at this stage is extremely common but the hyper-
trophied nucleus is generally not more basophilic than normal. A t a
later stage the basophilic material may become collected into a dense
mass which may continue t o occupy a position on the margin of the
nucleus, or again become more or less central. Still later, acidophilic
material may have disappeared, and all that remains of the nucleus
niay be one or niore fragments of deeply-staining basophilic chromatin
eriibedclecl in the liyaline cytoplasm of the rounded-off cell (fig. 8).
The seclnence of the nuclear changes particularly in the earlier stages,
runs parallel to that observed in yellow fever, and the acidophilic intra-
nnclear niasses develop in exactly similar fashion to the intranuclear
‘‘ inclusion bodies ” of Torres (1928), and Cowdry and Kitchen (1930).
The aciclopliilic material in this disease is more finely graindar tljnil
that in either yellow fever or herpes.

Discussion.
This clisease has undoubtedly existed in the Rift Valley for some
>.ears, and from time to time it has been responsible for extensive
losses both in adult sheep and in young lambs. No precise records
576 R. DAUBNEY AND /. R. HUDSON

are available, but according t o information obtained from stoclr-owners

in the district certain wet years have been noticeable for an excessive
number of abortions and a heavy mortality in adult sheep. I n other
wet years the chief feature has been the mortality in very young lambs.
I n each case there has been a history of abortion in cattle and of a
dengue-like fever in man, occurring a t the same time. The mortality
in lambs has generally been attributed to a bacillary infection, probably
to b’. eoli, and the mortality in ewes was thought to be due to heart-
water, a disease which before the discovery of Rickettsia presented
considerable difficulty in diagnosis. The dengue-like fever in man
would in all probability fail to attract serious attention in view of the
fact that in such years the incidence of malaria would be unusually
high. I n the year 1926 a t least three stock-owners suffered from a heavy
mortality among ewes and a high abortion rate. There is also a history
in this year of numerous abortions among cattle and tho occurrence of
a clengue-like fever of man both among natives and Europeans. The
incidence of malaria in 1926 was the highest for some years.
I n the light of our experience the two chief factors concerned in
the natural control of the disease appear to be the lower susceptibility
of adult sheep and the clistribution of the vector. The nunibers of
the vecting insect appear to be controlled by climate, which operates
hy limiting its distribution both geographically and seasoiially. If
lambing is taking place when and where the vector attains its peak in
numbers, heavy mortality among young lambs is to be expected. If on
the other hand it is possible to arrange that lambing shall fall a t a
time when the vector is not normally active, or if it is possible to lamb
at higher altitudes where the vector does not occur, mortality may be
greatly reduced.
I n the light of these facts it has been possible to prevent serious
outbreaks on the farms in the vicinity of the original outbreak during
the present year. On one farm the disease caused mortality in the
ewe flocks during the whole of the period J u n e to September. Before
lambing in November the flocks were moved to higher country and
no cases of the disease were reported. On another farm the disease
was responsible for mortality in ewes and rams. I n a small rain flock
there was a mortality of roughly 30 per cent. during August and
September; and in one of the ewe flocks some forty abortions were
reported, while an examination of the ewes indicated that a much
larger number of abortions had actually occurrecl. Diagnosis was
confirmed by bloocl inoculation, and although lambing was timed to
commence within three weeks the owner was aclvised to move the
ewes to a higher farm. Only two or three cases of the disease were
diagnosed in the lambs born after movement. A similar movemeilt
was carriecl out from the third farm with equally successful results.
Here a few ewes were retained to lamb a t the home farm, and in
their lambs a mortality in excess of 50 per cent. occurred.
 RIFT VALLEY FEVER 677

We are unable to trace any reference to this disease or to any

similar disease in literature or in the records of this laboratory; and
differential diagnosis from the better-known sheep diseases offers no
difficulty. The four common virus diseases of sheep in Kenya are
Nairobi sheep disease, bluetongue, heartwater and sheep-pox. Sheep-
pox is readily distinguished as a member of the variola-vaccinia group.
The incubation period in Nairobi sheep disease is slightly longer than
that of Rift Valley fever, and the disease is not transmissible to
bovines. Moreover the chief symptom in Nairobi sheep disease is
diarrhea, and at death characteristic lesions are found in the stomach,
czcum, colon and kidneys. The incubation period of bluetongue is
from seven to ten days after blood inoculation and in a natural out-
break the mouth lesions are characteris tic. Heartwater has a similar
incubation period to bluetongue and here again the lesion from which
the disease takes its name is found in a percentage of cases. I n
Nairobi sheep disease, bluetongue and heartwater tumor plenis is a
constant post-mortem lesion while it has only been observed once in
E f t Valley fever. Animals known to be immune t o all these diseases
were frequently employed in the early transmission experiments, and
almost every animal inoculated with the virus of enzootic hepatitis
has also been used either before or afterwards in Nairobi sheep
disease experiments without any evidence of cross immunity being
detected.
The only disease of cattle with which Rift Valley fever niight
possibly be confused is three day sickness, which has been variously
referred to in literature as three day sickness, s tiff-sickness, ephemeral
fever, and dengue. Three day sickness of cattle in Africa is a niild
inoculable febrile disease of short duration, characterised by muscular
and articular pains and stiffness. Affected cattle are invariably lame
in one or more limbs and may be so badly affected as to be unable to
rise. The illness lasts from three to five days and is followed by a
rapid and complete recovery. There are records (unpublished) in
this laboratory of certain experiments carried out by Montgomery,
in which the incubation period of three day sickness after blood
inoculation is shown to be from five to eight days, as against three
to five days for enzootic hepatitis in cattle. Moreover, Montgomery
was unable to transniit three day sickness to sheep, although some
twenty sheep were inoculated on various occasions with blood con-
taining the virus.
Clinically three clay sickness is a well defined and easily recognisecl
disease. It is relatively common in certain years in Kenya Colony and
does not differ in any way frorii the disease described by responsible
workers in other parts of Africa. Unfortunately, the name “ dengue ”
has been applied to this disease of cattle by various authors. Although
there is some uncertainty as to the identity of the disease with which
some of the older authors were working, the “dengue” of Rot-Bey
JOURN. OF PATH.-VOL XXIUV. 2 Q
578 R. DAUBNEY A N D /. R. HUDSON

(1895), Hutyra and Marek (1916) Edmonds and Walker (1929) and
Tarantino (1929) is undoubtedly the disease known to us as three day
sickness.
The true dengue of man is not transmiseible by inoculation to
cattle (Melanicli and Stylianopoulo 1930). We can trace no records
of attempts to infect sheep. This brings us to the question of the
identity of Rift Valley fever as observed in man with true dengue;
and here, in addition to the difference in behaviour of the viruses in
cattle, the absence of a rash in Rift Valley fever iu man is of assistance
in distinguishing the diseases by clinical means.
The clinical resemblance of the disease in man to true dengue is of
particular interest when considered together with the remarkable
agreement histologically between the liver lesion in sheep affected with
enzootic hepatitis and that found in yellow fever in man. If our
tentative conclusion, that Tceniorhynchus transmits enzootic hepatitis,
is correct, this is additional evidence of the relationship of the viruses;
especially since it has recently been shown that T. africanus can
transmit yellow fever under experimental conditions (Philip 1930).
These three diseases seem to fall into a natural group.
We have proposed the name Rift Valley fever as a popular
alternative to our first suggestion cnzootic hepatitis, which was originally
applied to the disease in sheep. The latter is hardly a suitable name
for the human disease, since we have as yet no evidence that the liver
is involved in man. While realising the disadvantages of attaching a
geographical name to a disease, we feel that convenience necessitates
the use of some such term.

SUMMARY.
1. A virus disease affecting sheep, cattle, goats and man is described
from the Rift Valley of Kenya Colony.
2. The virus causes a mortality of over 95 per cent. in very young
lambs, abortions and a moderate mortality in ewes and cows and a
transient fever accompanied by severe pains in the region of the joints
in man.
3. In the susceptible domesticated animals the characteristic lesion
is found in the liver. Histologically this lesion is a focal necrosis,
bearing a striking resemblance to the Councilman lesion of yellow
fever.
4. The virus passes regularly through Chamberland filters up to the
L 11 grade, and occasionally through the L 13 candle.
5. The disease is not contagious and there is evidence to suggest
that it is transmitted by a mosquito, probably Tcewiorhynchus
brevipalpis.
6 . It is suggested that the disease falls into a natural group with
yellow fever and dengue in man.
 RIFT VALLE Y FE VER b79

REFERENCES.
COWDRY,E. V., AND KITCHEN,Amer. Journ. Hyg., xi. 227.
8. F. (1930)
EDMONDS, C. R., AND WALKER, ‘‘ Diseases of Animals in Tropical Countries,”
G. K. (1929) p. 382.
HUTYRA AND MAREK (1916) . Spec. Path. and Therap. of Dis. of Domestic
Animals. English Edition, i. 208.
KLOTZ,O., AND BELT, T. H. Amer. Jour.. Path., vi. 663.
(1930)
LAUDA,E., AND LUQER,A. (1926) Ergebn. in. Med. Iiinderhlk., xxx. 377. Quoted
by Ludford, R. I., in X e d . Res. Council’s
System. Bact., vii. 40.
MELANIDI,C., AND STYLIANO- Rev. Gen. Ned. Vet., xxxix. 640.
porno, M. (1930)
OLITSKY,P. K., AND BOEZ,L. Journ. Exp.ilfed., xlv. 685.
(1927)
PEILIP,C. B. (1930). . . . . Amer. Journ. Trop. Xed., x. 1.
PKOT-BEY (1895) . . . . . . quoted Hutyra and Marek, vide aupa.
SIMMONS, J . S., ST JOHN,J . H., Phillip. Journ. Sci., xliv. 1.
AND REYNOLDS,F. H. K. (1931)
TARANTINO, G. B. (1929) . . . La clinica vet., lii. 77.
TORRES (1928). . .. . . . C.3. SOC.Biol., xcix. 1344.

JOUBN. OF PATH.-VOL. XXXIV.