Path. vet.

I: 504-529 (1964)

From the Department of Veterinary Preventive Medicine

University of Queensland, Brisbane

Studies on the Pathology of Experimental Lantana

(Lantana camara L.) Poisoning of Sheep
A. A. SEAWRIGHT

In surveying the history of experimental intoxication of cattle and

sheep by feeding the plant Lantana camara, SEAWRIGHT~~ pointed out
that in prior reports, only gross pathological findings were described.
In a study in which 25 sheep were poisoned, resulting in five deaths,
SEAWRIGHT reported that with the exception of the kidney, few
significant histopathological changes were observed. In the livers of
some sheep, marked centrolobular accumulation of bile pigment in
parenchymal and Kupffer cells was all that could be observed. In
sheep which died when hepatic dysfunction had apparently disap-
peared, no significant histopathological changes could be observed in
the liver.
SASTRYand MAHADEVAN~~ reported studies on experimental poi-
soning of sheep with “lantadenes” and while hepatic lesions were
reported as being present, no description of the histopathology of the
affected organs was given.
TOAHEIKEL et al.35 used rehmannic acid extracted from lantana
leaf in studies on bile flow in the rabbit. While paralysis of bile flow
could be produced with intraperitoneal administration of the suspend-
ed finely powdered crystals, affected livers showed no significant histo-
logical changes.
HANZON had
~ ~shown in studies on the dynamics of bile flow in
normal and intoxicated mouse liver that it was possible to halt bile
flow from cells which were histologically normal. TOAHEIKEL et a1.35
and SEAWRIGHT~~ then accepted that lantana plant (or lantana derived
rehmannic acid) poisoning were instances of the so called “intra-

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 S E A W R I G H TLantana Poisoning of Sheep 505

hepatic cholestasis” in which the excretory function of the liver was

deranged without demonstrable morphological damage to the paren-
chyma or obstruction in the biliary tract.
The terminal lesions in the kidneys in the special case of apparent
resolution of hepatic dysfunction before death have been described by
SEAWRIGHT~~ as nephrosis. However, there was usually a transient
proteinuria earlier in the syndrome which corresponded to the period
of apparent hepatic dysfunction. The terminal renal lesion was then
considered to be probably secondary and perhaps associated with
prolonged gut stasis, a view originally put forward by SEDDON and
CARNE~O and recently shared by JANSEN and ALEXANDER~~. Since all
the animals in that study were allowed to follow a normal course to
recovery or death, there was no opportunity to examine the kidneys
during this initial period of proteinuria. The tendency to dehydra-
tion through excessive loss of water, mainly as urine, during the first
week after intoxication was also observed and this may have had its
origin in renal tubular dysfunction.
BROOKS~ in describing lantana poisoning of cattle under field con-
ditions reported that in the case of hungry cattle, mortalities were
known to occur within 48 hours of eating the plant. In such cases the
animals showed symptoms of abdominal pain. S A N D E R S described
~~
gastroenteritis in cattle fed lantana leaves.
Accordingly, an experiment was designed specifically to study the
morbid anatomical changes in sheep poisoned with Lantana camara,
with particular reference to lesions of the liver and kidney.

Methods and Materials

Ten castrated male lambs averaging 20 kg in weight were used in

this study. They were accommodated in sheep metabolism cages
which were kept in a shed so that the sheep were not exposed to
direct sunlight. Feed and water intake and faeces and urine output
were measured for each animal.
In the cases of lambs numbered 257, 259, 256, and 254, a single
dose of toxic powdered lantana leaf (BRI 038434) at 6g/kg of body
weight was given as previously described (SEAWRIGHT~~). Since in
cattle under field conditions, poisoning seems to be most severe when
the animals are hungry and thirsty, the remaining six lambs, numbers
243,244,245,236,237, and 238 were given doses of powdered leaf at

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506 SEAWRIGHTLantana Poisoning of Sheep

Table I. Pretreatment, Dosage arid Post Dosage Procedure for each Sheep
Sheep Total dose Post-Intox. Predose Period Haematocrit Total Serum
No. g/kg B.Wt. Period without food at Death Bilirubin
Days and water. mg% at
Hours death
257 6 1 - 33 1.2
259 6 2 - 33 3.7
256 6 6 - 34 6.9
254 6 5* - 32** 7.1
243 12 7 24 38 14.6
244 12 7 24 45 19.4
245 12 30 * 24 30 15.3
236 12 7* 48 65** 15.2
237 12 7* 96 67** 10.4
238 12 14* 48 35 15.3
* Died or killed in a moribund condition. The remainder were destroyed for
the purpose of studying the progress of lesions.
** Sheep 254 and 236 died before a blood sample could be collected and the
sample for 237 was accidently lost. These readings are those on samples
collected 24 hours previously.

the rate of 6 g/kg of body weight on each of two consecutive days after
varying periods of 24,48, and 96 hours deprivation of food and water,
as the case may be, (Table I). With each animal, daily determinations
were made of urinary bile pigment, protein, and specific gravity.
Blood samples were collected daily and the haematocrit and total
serum bilirubin measured. When animals died or were destroyed at
predetermined periods, necropsies were performed as soon as possible
after death. The volume and colour of gall bladder contents were noted
as was the volume of urine in the bladder. Urine samples were collected
for determinations of bile pigment, protein and specific gravity.
The right adrenal gland was dissected out in each case and weigh-
ed. Slices of the left adrenal gland in each case were fixed in 10%
buffered neutral formalin and Helly's fluid. Slices of liver, kidney,
heart, lung, thyroid, pituitary, rumen, reticulum, omasum, abomasum,
duodenum, duodenum including the ampulla of Vater, ileum, colon,
rectum, gall bladder, cystic duct, hepatic duct, pancreas, and mesenteric
and precrural lymph nodes were also fixed in 10% buffered neutral
formalin. Slices of liver, kidney, pancreas, pituitary, adrenal, thyroid
and gall bladder were fixed in Helly's fluid. Slices of liver were fixed
in cold acetone at 4" C for histochemical localisation of alkaline phos-

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 S E A W R I G H TLantana Poisoning of Sheep 507

Table 11. Adrenal Weights, Amount and Colour of Bile, and Specific Gravity,
Protein and Bile Pigment Content of Urine at Necropsy

Bile Urine

Sheep Rt. Vol. Colour Bile Protein Specific

No. Adrenal ml Pigment mg/ml Gravity
Wt. (g) (&I
257 1.5 30 Darkgreen - 0 1.035
259 1.33 45 Darkgreen - 0 1.030
256 1.26 60 Green + 1.3 1.038
254 1.83 68 Green + 3.2 1.040
243 2.95 120 Pale green + 3.2 1.035
244 2.35 240 Palegreen + 3.2 1.032
245 2.98 70 Darkgreen + 2.0 1.020
246 2.50 150 Colourless + -* -*
237 2.55 100 Darkgreen + 10 1.030
238 2.85 200 Colourless + 10 1.030
* This animal was dead 12 hours or so when necropsy was performed and apart
from examination for the presence of bile, testing was not carried out.

phatase. In the case of livers of sheep 257, 259, 256, 243 and 244,
samples of the organs were fixed in 1% buffered osmic acid for electron
microscope studies. Blocks of fresh tissue from heart, kidney and
liver were frozen at -20” Cfor later cryostat sectioning. Paraffin sections
were prepared from all tissues fixed in formalin and Helly’s fluid and
stained with Harris’s haematoxylin and eosin. Frozen sections were
prepared from formalin fixed and fresh frozen liver, kidney and heart
and stained with Sudan Black B, Sudan Black B and carmalum, Sudan
IV and Haematoxylin, Oil Red 0 and Haematoxylin. Thin frozen
sections of 6,u were cut on a cryostat microtome and thick frozen
sections of 10-15 p on a conventional freezing microtome. Helly’s
fixed paraffin sections of kidneys were stained with azan, and formalin
fixed paraffin sections of kidneys were stained with Von Kossa’s stain
to demonstrate calcium deposits. Helly’s fixed paraffin sections of
gall bladder were stained with Weigert-Hart and azan. Formalin fixed
paraffin sections of liver were stained with P.A.S., P.A.S. with dias-
tase digestion, azan, Van Gieson, toluidine blue, Southgate muci-
carmine, Foot’s reticulin, and HALL’+method for bile pigment, and
Helly’s fixed paraffin sections of liver were stained with phospho-
tungstic acid haematoxylin (P.T.A.H.).

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 508 SEAw R I G H T Lantana Poisoning of Sheep

Fag. 1. Liver of sheep 244. Swelling and paleness of staining of peripheral paren-
chymal cells and centrolobular cells apparently unaffected-slight proli-
feration of biliary ductules and increase in mononudear mesenchymatic
cells (central vein, C; portal vein, P). H & E x 100.
Fig. 2. Liver of sheep 244. Peripheral parenchymal cells and irregularly shaped
densely staining smaller parenchymal cells (arrows). H & E x 500.

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 S E A W R I G H TLantana Poisoning of Sheep 509

Results

The results to be reported and discussed in this section are

pathological findings and only such clinical observations as have a
direct bearing on the pathological findings. Studies on the histo-
chemical localisation of alkaline phosphatase in the livers of these
sheep and those dealing with electron microscopy will be reported
separately.
Details of dose rate of toxic leaf, predosage treatment, duration
of intoxication, terminal haematocrit and total serum bilirubin are set
out in Table I.
Within twelve hours of poisoning, sheep 257, 259, 256 and 254
stopped eating and for the first seven days after, all animals drank less
water each day than they passed as urine. Little or no faeces was
passed after the second day and clinical jaundice was apparent in all
sheep after three days.

Gross Pathological Findings

Details of right adrenal weights, volume and colour of bile, and

determination of bile pigment, protein and specific gravity of urine
for each sheep at necropsy are set out in Table I1 and the main gross
pathological findings in Table 111.
Except for the animals destroyed within 48 hours of poisoning
most sheep showed jaundice and photosensitization, swollen, ochre
coloured and mottled livers, distended gall bladders with pale green to
colourless bile and moist kidneys. Acute constipation was indicated by
balling of dry faeces in the colon anterior to the ansa spiralis and
excessive mucus in the remainder of the large intestine. Most severely
affected animals showed excessive fluid in the body cavities and severe
pulmonary oedema.

Abb. 1. Schwellung und schwache Farbung der peripheren Parenchymzellen;

zentrolobulare Zellen offensichtlich nicht betroffen. Geringgradige Proli-
feration der Gallengange und Vermehrung mononukkarer mesenchyma-
ler Zellen. (Zentralvene = C, Pfortader = P).
Abb. 2. Periphere Parenchymzellen und unregelmassig geformte, stark gefarbte
kleinere Parenchymzellen (Pfeile).

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 Table 111.Gross Pathological Findings m
+
0

Sheep Jaundice Liver Kidney Lower alimentary tract Heart Lungs Other findings
No* (J)
Photo-
sensit. (P)
v,
M
257 - - - - - - - *
d
- Slightlypale - -
T1
259 - - - 0
z
and swollen +I

256 J Slightly swollen - - - - - F

P Normal colour g
254 J Swollen and Cut surface Large, dry bolus like - - -
2%.
P ochre coloured very moist faecal accumulation in E3”
proximal colon op
%
Swollen, bile Pale and bile Large, dry bolus like - - - v,
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243 J 5
P stained and stained faecal accumulation in 4
mottled proximal colon

Swollen but of - Rumen contents very dry - - -

244 J
normal colour and colon contained a
large faecal bolus
245 J Swollen and Very bile Colon contained large Veryflabby - Gall bladder wall
P ochre coloured stained faecal bolus and distally and fat of appeared thicker
large intestine contained coronary groove than normal
excessive amount of grey, showed serous
opaque mucous atrophy
236 J Swollen, Dark red and Colon contained large, - Dark red, 200 cc blood stain-
P mottled, bile friable dry faecal bolus congested, ed pleural fluid. m
hi
stained moist, with 500 cc clear yellow
much white fluid in peritoneal 2
TI
froth in cavity 0
T
bronchi and +I
trachea
- r
237 J Swollen and Very pale with Stomach and small intes- Pale and Firm con- Subcutaneous and p,
clay coloured red striations tine contained copious flabby sistency, red intermuscular
with scant in cortex and amount of fluid. Few patches. tissues showed
patchy redden- medulla petechial haemorrhages in Moist on extensive petechial 3
ing under mucosa of duodenum and cutting with haemorrhages-
capsule ileum. Caecum and colon copious peritoneal cavity (19
E'
contained masses of hard, white froth contained 300 cc
%
dry, black faeces in trachea clear yellow fluid V,
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and bronchi a
238 J Swollen and Swollen, bile Anterior colon contained Twelve haemor-
4
P bile stained stained large, dry, black bolus of rhagic ulcers
with pale faeces 2-5 mm in diame-
mottling of ter on gall bladder
parenchyma mucosa
(P) = Inflammation of the conjunctivae and the skin of the muzzle and tips of the ears (see CLARE?).
512 S E A W R I G H TLantana Poisoning of Sheep

Histopathologic Findings

Liver

After 24 hours of intoxication the only change that could be ob-

served in the liver was an increase in the intensity of staining of the
cytoplasm of the peripheral parenchymal cells and an increase in the
granularity of the cytoplasm of more centrolobular cells. After 48
hours, the peripheral parenchymal cells showed a slight degree of
swelling with a tendency to paleness of the cytoplasm and prominence
of bile canaliculi. After 5 to 6 days there was very marked swelling of
periportal parenchymal cells, and their cytoplasm was quite pale and
granular. At this stage, there was a slight increase in collagen and
reticulin fibres in portal tracts and a slight tendency to bile staining of
the cytoplasm of more centrolobular parenchymal cells. Some glycogen
remained in periportal parenchymal cells and after diastase digestion,
P.A.S. positive dots, granules and globules were present in some more
centrolobular parenchymal and Kupffer cells.
From 6-7 days after poisoning and especially in sheep in which
the condition was much more severe, the periportal parenchymal cells
showed further degrees of degeneration. (Fig. 1). The cytoplasm
became very pale and vacuoles could be seen in some cells. Yellow
injected bile canaliculi stood out clearly and fine lines of bile pigment
appeared almost completely to surround parenchymal cells in the
periportal region. In these areas of the lobules there were numbers of
small irregularly shaped parenchymal cells with very eosinophilic and
homogeneous staining cytoplasm and small very dense nuclei (Fig. 2).
These latter cells appeared to be intimately associated with adjacent
parenchymal cells. The next stage of the degeneration of periportal
parenchymal cells was marked swelling of the cells with extensive
vacuolation of the pale staining cytoplasm and pyknosis of many
cell nuclei (Fig. 3). The nuclei in many hepatocytes situated in peri-
portal and midzonal regions showed a characteristic “bird‘s eye” like
appearance with a thin well-defined nuclear membrane and the chro-
matin accumulated into a large dense centrally placed dot. The
vacuoles in the cytoplasm of these cells were sometimes large and
single but more often were multiple so as to give the cytoplasm a
lacelike or foamy appearance. The vacuoles were only partially due to
fat, Some globules or granules of bile pigment were also observed
in such cells. At this stage, the cells surrounding the central vein

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 S E Aw R I G HT Lantana Poisoning of Sheep 513

3 4

Fig. 3. Liver of sheep 243. Periportal vacuolar degeneration of parenchymal cells

and apparently normal centrolobular cells. Marked biliary ductular hyper-
plasia (central vein,C; portal vein, €'). H & E x 150.
Fig. 4. Liver of sheep 237. Extensive necrosis and vacuolation of parenchymal
cells with proliferation of bile ductules. Considerable apparent regenerative
activity of parenchymal cells with disorganisation of the lobular archi-
tecture (central vein, C; portal vein, P). H & E x 150.

A b b . 3. Periportale vakuoliire Degeneration der ParenchymzelIen und offen-

sichtlich normale zentrolobulare ZelIen. Starke Hyperplasie der Gallen-
gange. (Zentralvene = C, Pfortader = P).
Abb. 4. Ausgedehnte Nekrose und Vakuolisierung der Parenchymzellen mit Proli-
feration der Gallengange. Sichtlich starke regenerative Aktivitat der
Parenchymzellen bei Auflosung der lobularen Struktur. (Zentralvene = C,
Pfortader = P).

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514 S E A W R I G H T Lantana Poisoning of Sheep

Fig. 5. Liver of sheep 245. Sporadic centrolobular and midzonal vacuolation

of parenchymal and Kupffer cells of long standing hyperbilirubinaemia.
Changes in periportal parenchymal cells and portal tracts are less apparent
(central vein, C ; portal vein, P). H & E x 150.
Fig. 6. Kidney of sheep 254. Extensive vacuolar degeneration of the cytoplasm
of the cells of the uriniferous tubules with pyknosis and disappearance of
nuclei in many cells. H & E x 150.

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 SEAWRIGHTLantana Poisoning of Sheep 515

appeared normal while the parenchymal cells immediately central to

the zone of vacuolar degeneration showed a marked increase in the
granularity of the cytoplasm. At 7 days of poisoning there was no
glycogen apparent in the liver but there was an increase in the P.A.S.
positive and diastase resistant deposits in mainly centrolobular paren-
chymal and Kupffer cells. The most severely intoxicated sheep
(No. 237) was also destroyed after 7 days and the microscopic feature
of this liver was the extensive degrees of necrosis and disappearance
of parenchymal cells, and disorganization of the liver lobules. Residual
parenchymal cells showed considerable pleomorphism, decrease of
eosinophilia of the cytoplasm with a variable degree of vacuolation,
almost complete loss of P.T.A.H. staining of the cytoplasm and chroma-
tolysis and pyknosis of nuclei. Mitotic figures were plentiful in both
parenchymal cells, biliary ductules, and Kupffer cells (Fig. 4).
There was a marked degree of biliary ductular hyperplasia in all
livers by the seventh day after intoxication and an increase both in the
amount of collagen and reticulin fibres and in the numbers of mono-
nuclear mesenchymatic cells in the portal tracts and peripheral regions
of the lobules. There was some breakdown of reticulin fibres in the
region of damaged parenchymal cells. With extensiveparenchymal cell
necrosis there appeared to be an increase in the number of fibres pre-
sent within the lobule also. With severe intoxication there was also
marked distension of the lymphatics of the portal tracts with oedema,
hyperaemia, haemorrhage and mononudear inflammatory infiltra-
tion of the periductal connective tissues, and desquamation of bile
duct epithelium.
The more chronically affected livers were characterised by a con-
siderable increase in the degree of centrolobular and midzonal bile
pigment accumulation. This took the form of pronounced finely granu-
lar brownish staining of the cytoplasm of parenchymal cells and large

A b b . 5. Sporadische Vakuolisierung der Parenchym- und Kupfferschen Stern-

zellen, zentrolobular und im mittleren Bereich bei lang anhaltender
Hyperbilirubinamie. Veranderungen der periportalen Parenchymzellen
und Pfortaderverzweigungen sind weniger offensichtlich. (Zentralvene
= C, Pfortader = P).
Abb. 6. Ausgedehnte vakuolare Degeneration des zellularen Plasmas der Harn-
kanalchenepithelien mit Pyknose und Untergang der Kerne in vielen
Zellen.

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516 SEA w R I G HT Lantana Poisoning of Sheep

irregular granular and globular, brownish and P.A.S. positive deposits

sporadically located in vacuolated parenchymal and Kupffer cells. Bile
plugs also were plentiful. Peripheral parenchymal degeneration with
increased periportal fibrosis and ductular hyperplasia were still evident.
However, after 28 days, the changes in periportal parenchymal cells
were less apparent while sporadically vacuolated cells appeared to
have moved to midzonal and centrolobular regions (Fig. 5). In addi-
tion, many of the newly proliferated biliary ductular cells showed loss
of cytoplasm suggesting a tendency to involution.

Gall Bladder

After poisoning, the gall bladder became progressively distended

but at the lower dose rate and up till G days after dosing there were no
significant histological changes in the wall of this organ other than
slight interstitialoedema and distendedlymphatics.In the more severely
poisoned animals, where after one week, the gall bladders were
distended to 4 to 8 times normal, there was marked hyperaemia and
oedema with polymorphonuclear and mononuclear inflammatory cell
infiltration of all layers of the wall. The mucosa remained intact. The
wall showed atrophic and degenerativechangesin smooth muscle fibres,
thrombosis of some small arteries, and hyalinization and poly-
morphonuclear cell infiltration of the walls of some other blood
vessels.
When the intoxication was of longer standing and the degree of
distension still appreciable, there was marked increased thickness of
all submucosal layers of the organ due to increased deposition of
fibrous tissue. At this stage interstitial oedema was absent but there
remained a moderate degree of polymorphonuclear cell infiltration of
all tissues and a protein coagulum in distended lymphatics. The mu-
cosa remained unaffected except in one sheep (No. 238) where there
were about 12 haemorrhagic ulcers 2 to 5 mm in diameter.
There was no evidence of acute cholecystitis reported to occur in
lantana poisoning by SEAWRIGHT~~. Even the most acutelyaffected sheep
showed no inflammation of the gall bladder mucosa. It should also
be noted that cholecystitis has been rarely seen in lantana poisoned
animals kept in metabolism cages, but it is not uncommonly seen
when the movement of the animal has not been so restricted. Since
it appears unlikely that the toxic principle of this plant has any effect

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 SEAWRIGHT
Lantana Poisoning of Sheep 517

on the gall bladder mucosa, it is suggested that the cause of acute

cholecystitis is due to mechanical factors related to the great degree
of distension of the gall bladder and opportunity for movement by
the animal. It is further suggested that by sheer weight a distended gall
bladder would kink with frequent violent movements-as in walking
and running-and so produce interference with the blood supply to the
organ, giving rise to local areas of ischaemia of the mucosa. This is
supported by the fact that acute cholecystitis is rarely, if ever, accom-
panied by infection and the changes are patchy in distribution rather
than involving the whole mucosal surface. The inflammatory changes
in the gall bladder wall are considered to be associated with distension
and stasis since the changes in lantana poisoning show no significant
differences from those seen when the common bile duct is obstructed
in non-poisoned sheep".

Kidney

The earliest change detected in the kidney was a slight increase in

the protein content in the fluid of the uriniferous tubules at 24 to 48
hours after poisoning. This was followed in 5-6 days by a slight
degree of fatty degeneration in the cells of the proximal convoluted
tubules. With increasing degrees of severity and time after poisoning,
the cells of the proximal convoluted tubules showed increased eosino-
philia with pyknosis of nuclei in some nephrons, while there was a
marked increase in the degree of non-fatty vacuolar degeneration of the
cytoplasm of other tubule cells (Fig. 6). The dense eosinophilic
cytoplasm of some epithelial cells broke down to bright red globules
and granules. With increase in severity there was frank coagulative
necrosis of tubular epithelium which in the most advanced case in-
volved most of the uriniferous tubules of the cortex, and even the
collecting tubules of the medulla. In such necrotic tubular cells, va-
cuoles could still be recognised while nuclei had disappeared and the
cells had desquamated from their basement membranes (Fig. 7). There
was also a further increase in the amount of bile stained protein con-
taining fluid in the lumina of uriniferous tubules and a tendency to
bright eosinophilic hyaline cast formation. Where necrosis of tubular
epithelium was present there was considerable infiltration of the
juxta-medullary areas with mainly mononudear and some poly-

* SEAWRIGHT,
unpublished.

42 Path. vet., Val. 1, No. 6 (1964)

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 518 S E A w R I G H T Lantana Poisoning of Sheep

Fig. 7. Kidney of sheep 237. Extensive necrosis of the tubular epithelium of the
cortex. The nuclei of many cells are pyknotic and vacuoles can be seen in
the cytoplasm of some cells. H & E x 150.
Fig. 8. Heart of sheep 238. Reparative lesion in the myocardium 14 days after
poisoning. The cellular areas contain mainly proliferating fibroblasts,
histiocytes and atrophic muscle fibres. H & E x 300.

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 S E A W R I G H TLantana Poisoning of Sheep 519

morphonuclear inflammatory cells. In areas of mainly necrotic

tubular epithelium in the cortex, blood vessels appeared engorged and
there were occasionalhaemorrhages. In all kidneys where the intoxica-
tion was of more than 5 days’ duration there was an increase in the
degree of interstitial calcium salt deposits in the medulla.
Similar degrees and types of degeneration of tubular epithelium
were present in the more chronically affected sheep. In these kidneys
however, there was more brown-stained proteinaceous fluid in tubules
and more extensive hyaline cast formation with consequent obstruc-
tive plugging of the tubular lumina. This gave rise to extensive disten-
sion of the uriniferoustubules with flattening of the tubular epithelium.
This was most evident in the outer third of the cortex where there
was a marked interstitial oedema and increased deposition of colla-
gen fibres. In such kidneys interstitial deposits of bile pigment were
also observed in the juxta-medullary areas.

Heart

Three of the most acutely affected animals showed microscopic

lesions involving the myocardium of the left ventricle when necro-
psied 7 days or longer after poisoning. The earliest changes observed
were patchy degeneration of heart muscle fibres with scanty petechial
haemorrhages. Affected fibres appeared fragile and misshapen with
loss of staining of some nuclei and loss of striation. There were some
mononuclear phagocytic cells located between affected fibres. In the
same heart, and in all others showing lesions, were multiple foci of scar
tissue consisting of histiocytes, proliferating fibroblasts and atrophied
muscle fibres (Fig. 8). In the most severely affected sheep however,
while no such degenerative or inflammatory changes were observed
in the myocardium, many petechial haemorrhages, interstitial oedema
and distended lymphatics were observed microscopically.

A b b . 7. Ausgedehnte Nekrose des tubularen Epithels in der Nierenrinde. Viele

Zellkerne sind pyknotisch und im Zytoplasma einiger Zellen kann man
Vakuolen erkennen.
Abb. 8. Reparative Veranderungen des Myokards 14 Tage nach Vergiftung. Die
zellulaien Gebiete enthalten hauptsachlich wuchernde Fibroblasten, Histio-
zyten und atrophische Muskelfasern.

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 Table IV.Histopathological Findings ul
E3
0

Sheep Liver Gall Bladder Kidney Heart Lungs

No.

257 Eosinophilia of periportal - - - - v,

m
parenchymal cells 5
z
0
259 Slight swelling of periportal Slight interstitial oedema - - - z
rl
parenchymal cells
r
256 Moderate hydropic change Slight interstitial oedema Slight fatty degeneration of - -
of periportal parenchymal proximal convoluted v
cells tubules g.
8
5'
254 Moderate hydropic change Slight interstitial oedema Marked vacuolar degener- - - m
h

of periportal parenchymal ation of tubular epithelium

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cells of cortex

243 Marked vacuolar degenera- Marked interstitial oedema, Moderate vacuolar de- Extensive patchy
tion of periportal paren- hyperaemia and inflamma- generation of tubular myocardial degener-
chymal cells, portal fibrosis tory cell infiltration of epithelium of cortex ation and slight
and ductular hyperplasia wall scar formation
244 Marked hydropic change Marked interstitial oedcma Slight vacuolar degener- - -
in periportal parenchymal hyperaemia, and inflamma- ation and cast formation in
cells, portal fibrosis and tory infiltration of wall uriniferous tubules
ductular hyperplasia

245 Slight periportal hydropic Marked fibrous thickening Moderate vacuolar degener- - -
change, marked fibrosis, of wall, moderate inflam- ation of tubules with cast
ductular hyperplasia and matory infiltration formation and extensive
sporadic centrolobular and plugging of tubules with
midzonal vacuolation and proximal cystic distension
bile pigment deposition

236 Marked periportal vacuolar Moderate interstitial Extensive necrosis and Slight degree of Pulmonary
degeneration, portal oedema and inflammatory vacuolar degeneration of scar formation in congestion
fibrosis and ductular cell infiltration all tubular epithelium myocardium and oedema
hyperplasia

237 Extensive necrosis of Moderate interstitial Extensive necrosis and Oedema and Pulmonary
parenchymal cells with oedema and inflammatory vacuolar degeneration of petechial haemor- congestion
marked regenerative cell infiltration all tubular epithelium rhage of myocard and oedema
activity biliary ductular ium
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hyperplasia

238 Marked periportal Marked fibrous thickening Slight vacuolar degener- Multiple scar
vacuolar degeneration, of wall, inflammatory ation, plugging of tubules formation in
portal fibrosis and infiltration, 12 small with casts and proximal myocardium
ductular hyperplasia ulcers on mucosa cystic distension
522 S E A W R I G H T Lantana Poisoning of Sheep

Onlyin the lungs of the twomost severelyaffectedanimalsandthose

which showed pulmonary oedema at the necropsy were lesions ob-
served. Microscopically, capillaries were greatly congested and alveoli
in many areas were filled with a homogeneous pink staining fluid.
In other regions alveoli were greatly distended, indicating emphysema,
and in others, contained scanty fragments of fibrin only. Peribronchial
and perivascular lymphatics as well as inter-lobular septa were greatly
distended and showed fragments of fibrin coagula in their lumina.
The principal microscopic changes in respect of each animal in
this series are set out in Table IV.

Discussion

Pluricausality of Lesions with Particular Reference to the Heart

S E L Y Eand
~ ~ BAJUSZ and SELYE~ have demonstrated with exhaus-
tive experimentation in the rat that a great variety of degenerative
and inflammatory lesions can be produced in the heart when the
animals are prestressed and conditioned in various ways and then given
certain potentially cardiotoxic drugs. In many cases, marked liver and
kidney lesions are observed as well. These experiments in sheep have
indicated that preconditioning by starvation and thirst prior to in-
toxication with lantana leaf gives rise to extremely severe lesions in
the liver and kidneys and also to degenerative and reparative lesions
in the heart. The situation here may well be comparable and the toxic
principle in lantana may be cardiotoxic as well as hepatotoxic and
nephrotoxic. In studies on the poisoning of rabbits with icterogenin,
a substance which produces a syndrome very similar to lantana poison-
ing, BROWN et al.2 claimed cardiac failure to be the cause of death in
the more acute syndromes. It would appear possible that in cases of
deaths from lantana poisoning in travelled or hungry and thirsty
cattle within 48 hours of exposure as claimed by BROOKS~, mortality
was probably due to cardiac failure. The latter may, in fact, be the
common cause of death when this occurs in the more acute phase of
lantana poisoning.

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 S E A w R I G H T Lantana Poisoning of Sheep 523

The Distribution and Nature of Primary Lesions

In the acute phase of lantana poisoning in these sheep the alimen-

tary tract, the liver and the kidneys appeared to be affected simul-
taneously. There is no evidence at present that this is also true of the
heart, although it seems likely to be so.
The intensity and duration of constipation in this condition
implies a paralysis of the lower alimentary tract. This often results
in such a prolonged period of anorexia that death can be simply
ascribed to starvation. RIMINGTON et al.25 showed that icterogenin
could cause such a cessation of movement in an isolated sheep in-
testinal strip, and SEAWRIGHT~~ suggested such an effect on the gall
bladder musculature might also be mainly responsible for the great
degree of distension which characterises that organ in this disease.

The significant change observed in the kidney in the acute phase

of the disease is a fatty and vacuolar degeneration of the uriniferous
tubules which, with increase in severity includes the collecting tubules
of the medulla as well. These tubular lesions which are also accom4
panied by proteinuria might conceivably account for the initial phase
of diuresis previously described by SEAWRIGHT~~. This in turn might
then result in a sufficient degree of dehydration to promote the
unusually marked absorption of water from the contents of the large
intestine and rumen which is apparent from post mortem findings in
this condition.

Liver

From these studies on the histology of poisoned livers, it is

evident that early degenerative change can be recognised in the peri-
portal parenchymal cells. This takes the form of an initial hydropic
degeneration involving the outer rim of cells, and with increasing
severity, spreads to involve additional more centrally located cells. The
degeneration proceeds to extensive vacuolation of the cytoplasm of
the affected cells, and in the most severe, to widespread necrosis of
most parenchymal cells. The serum bilirubin levels (Table 11) show,
nevertheless, that slight degrees of morphologic change are accom-

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524 S E Aw R I G HT Lantana Poisoning of Sheep

panied by appreciable degrees of hepatic excretory dysfunction. Very

severe photosensitization can then result with very little morphological
indication of hepatocellular damage. This often happens in natural
cases of the disease in cattle.
With more chronically affected sheep, changes in these cells be-
come less apparent and there is a tendency to sporadicvacuolation of
centrolobularly located cells and with accumulation of bile pigment.
Prolonged periods of hyperbilirubinaemia as seen in this disease are
frequently accompanied by sporadic “feathery” vacuolation of the
cytoplasm of parenchymal cells (POPPER and SZANTO~~). Hou et al.13
have demonstrated the toxicity of bile for cells and POPPERet a1.21
have demonstrated the P.A.S. positive, diastase resistant nature of
such altered hepatic biliary deposits which are so frequently seen in
such circumstances.
After about one week of intoxication it is not unusual to observe
increased mitotic activity in parenchymal cells in the periphery of
the liver lobules. Since this region normally contains the cytogenic
parenchymal cells RAPP PA PORT^^), loss of parenchyma in this region
at the periphery of the lobule would normally be expected to result in
vigorous regenerative activity. The histological findings in liver of the
most severely affected sheep (No. 237) in this study are in part due to
such attempts at regeneration.
Biliary ductular hyperplasia is consistentlyseen in this condition if
the intoxication is sufficientlysevere or prolonged. Proliferation of bile
ductules is frequently observed in pathological conditions of the liver,
particularly those associated with biliary obstruction and regenerative
hepatocellular phenomena and also as a primary response to alpha
naphthyl-iso-thiocyanate administration (LOPEZand MAZZANTI~~). In
lantana poisoning, considering the degenerative changes which occur
in peripheral parenchymal cells, the biliary ductular reaction is more
likely to be secondary to such changes rather than a direct result of
the toxin on the bile ductule epithelium.

The Liver in other Photosensitivity Diseases

The degenerative changes in the peripheral parenchymal cells of

the liver have been described previously in photosensitivity diseases
~ ~PLOWRIGHT
in cattle and sheep. W I L S O Nand et al.20 described such
changes in the livers of cattle showing delayed dimidium bromide

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 S E Aw R I G HT Lantana Poisoning of Sheep 525

poisoning and these lesions have been studied more recently in cattle
by FORDand B o Y D ~The ~ . latter workers report they were unable to
produce the lesions experimentally in any other species tried, other
than adult cattle. Such changes were seen in the livers of sheep
poisoned by Myoporm Zaetgm leaves and extracts (CUNNINGHAM and
HOP KIRK^) and by Myoporm acczminatmv (RIEKand WRIGHT~~). Recent
studies on sporidesmin poisoning of sheep by MORTIMER et al.18 have
shown that the early hepatic lesion in this condition is vacuolar degen-
eration of the peripheral parenchymal cells. BULLSin a survey of
hepatotoxic diseases of livestock indicated that the histological change
in the liver in lantana poisoning was in the portal tracts but gave no
further details.
MATHEwSI~in his report of Agave /echegai%?apoisoning of sheep
found liver and kidney lesions resembling those described here in more
chronic lantana poisoning. BROWN et al.3 studying the pathology of
geeldikkop of sheep in South Africa also reported the presence of
similar liver and kidney changes as those seen here in chronic lantana
poisoning of sheep. Other authors describing the pathological features
of photosensitivitydiseaseshave indicated that liver and k i h e y lesions
were present, but did not indicate their nature CLARE^). RIMINGTON
et al.25 showed that icterogenin isolated from Lippia rehmanni caused
a disease in sheep similar to lantana poisoning and reported that no
lesions could be seen microscopically in the liver.
In relation to the nature of,the basic liver lesions seen in these
conditions there is ample evidence that the parenchymal cells of the
h e r lobule have functional differences (STONERand MA GEE^^ ;
WILSON~~; NOVIKOFF~~; and SHANKet a1.32) and it would appear that
the poisonous principles in the diseases listed here belong with
phosphorus WILLC COX^^) and ally1 formate (ROSINand DOLJANSKI~~)
in chiefly affecting the peripheral parenchymal cells.

Secondary Lesions and Disturbances of Function

Although SANDERS~~ and STEYNand VAN DER WALT%report

subpleural haemorrhages in the lungs of experimental poisoning of
cattle and sheep respectively, there is no evidence from this study of
the lungs that there is any primary lesion of this organ which would

43 Path. vet., Vol. 1, No. 6 (1964)

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526 S E A W R I G H T Lantana Poisoning of Sheep

explain the development of a fatal pulmonary oedema particularly as

late as 7 days after intoxication. The terminal haemoconcentration ob-
served in affected sheep (Ht = 65, 67%) was undoubtedly secondary
to pulmonary oedema (FLOREY~). Although some degree of haemo-
concentration (Ht = 50%) does normally occur during the initial
diuretic phase of the intoxication (SEAWRIGHT~~), it does not approach
this level. While some increased viscosity of the blood would un-
doubtedly contribute to circulatory difficulty in the lungs, it seems
most likely that since myocardial lesions have been demonstrated, the
pulmonary oedema is secondary to failing heart action (DALEY et al.8).

Kidney$

It is notable that extensive coagulative necrosis of tubular epi-

thelium was observed only in the kidneys of the two sheep (Nos. 236,
237) which experienced terminal fatal pulmonary oedema. Necrotic
tubules frequently also showed residual vacuolar degeneration of the
cytoplasm. This would imply that the coagulative necrosis with
engorgment of contiguous blood vessels occurred subsequently to
the initial degenerative change. Considering the prolonged period of
severe haemoconcentration it seems most probable that these necrotic
changes were secondary and of ischaemic origin. The liver of sheep
237 also showed coagulative necrosis of apparently already degener-
ated cells and the ischaemia of terminal haemoconcentration may also
have played a part in the production of the advanced changes seen in
this organ.
It appears highly likely from this and previous studies (SEA-
WRIGHT~~) that there is a marked tendency to dehydration and an-
hydraemia in lantana poisoning. MARRIOTT~~ has shown that in an-
hydraemia, secretion of urine became decreased and the urine con-
tained protein and casts and its specific gravity was usually raised.
This was seen particularly in the most chronically affected animal, the
terminal urine of which also contained a trace of glucose. This was a
feature of dehydration also noted by MARRIOTT~~ and observed also in
similar naturally occurring diseases of sheep by JANSEN and ALEX-
ANDERl4.
In lantana poisoning of long standing, the histopathology of the
kidney is complicated by the presence of an initial degeneration of
tubular epithelium followed by the effects of secondary anhydraemia.

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 SEAWRIGHTLantana Poisoning of Sheep 527

The latter contributes to the excessive production of hyaline casts with

consequent obstruction of uriniferous and collecting tubules. This
may lead to urinary retention with elevation of blood urea as previ-
ously reported (sEAWRIGHTzg).

Szimmay
A study was made of experimental poisoning of 10 sheep given powdered
lantana leaf at 6g/kg and 12g/kg body weight after periods up to 96 hours of
deprivation of food and water. Jaundice and photosensitization were evident in
almost all animals after three days. At necropsy the most severely affected sheep
showed necrosis of the liver and kidney, and pulmonary oedema. Less severely
affected sheep showed vacuolar or hydropic degeneration of the hepatic peripheral
parenchymal cells, bile ductule hyperplasia and slight portal cirrhosis while centro-
lobular cells remained normal. Myocardial necrosis and scarring were present in
the hearts of some sheep. Hydropic and fatty degeneration of the tubular epithe-
lium of the kidneys was seen in the acute phase and Cast formation, occlusion and
cystic dilatation of proximal tubules were present in the more chronic stages
associated with dehydration.

Zxrammenfasswzg
Zehn Schafe wurden experimentell durch Verabreichung von 6 und 12g/kg
Korpergewicht Lantanablattern (Lantana camara L.) in Pulverform vergiftet,
nachdem den Tieren vorher bis zu 96 Stunden Wasser und Futter entzogen war.
Bei fast allen Tieren wurde nach 3 Tagen Gelbsucht und Photosensibilisierung
sichtbar. Die autoptischen Befunde ergaben bei den am schwersten betroffenen
Schafen Nekrosen von Leber und Nieren sowie Lungenodem. In den leichteren
Fallen wurde vakuolare Degeneration oder hydropische Schwellung der peri-
pheren Zellen des Leberlappchenparenchyms, Hyperplasie der Gallenginge und
leichte Zirrhose beobachtet, wahrend die zentralen Leberlappcheneellen unver-
sehrt blieben. Bei einigen Schafen fanden sich Myokardnekrose und Narben-
bildung im Herzen. Im akuten Krankheitsstadium wurden hydropische und fet-
tige Degeneration des tubularen Nierenepithels festgestellt, wahrend in den von
Dehydration begleiteten mehr chronischen Stadien Zylinderbildung, Verschluss
und zystische Dilatation der proximalen Nierentubuli vorhanden waren.

Acknowledgements
I wish to acknowledge the continued support and advice of Professor
J. FRANCISduring the currency of these studies. The histological technical work
was performed by Mr. F.L. WEBBof this Department and Mr. A. DRAYDON of
the Department of Anatomy, University of Queensland. The photomicrography
was done by Mr. E. HOLLYWOOD of the University of Queensland Photographic
Department. Mr. D. W. W. JONES assisted me in the post mortem room and my
technical assistant, Mr. F. G. SFERCO assisted me in all phases of the work.

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528 SEAWRIGHTLantana Poisoning of Sheep

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