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Vet Pathol32:635-640 (1995)

The Lesions of Hepatic Fatty Cirrhosis in Sheep


R. G. HELMAN,L. G. ADAMS,
AND C. H. BRIDGES

Department of Pathobiology, Texas Veterinary Medical Center, Texas A&M University,


College Station, TX 77843-4467

Abstract. During a natural outbreak of hepatic fatty cirrhosis (HFC) in western Texas, 500 2-6-year-old
Rambouillet ewe sheep were sequentially studied to determine the pattern of lesion development. All sheep
developed lesions of HFC. Grossly, changes first began in the subcapsular hepatic parenchyma along the porta
hepatis and spread peripherally until, in the final stages of the disease, approximately 80% of the liver was
affected. Ascites, hydropericardium, and acquired hepatic vascular shunts were present in sheep with severe
HFC. Light microscopic lesions initially appeared as accumulations of fine lipid droplets in the cytoplasm of
periacinar hepatocytes but, with time, involved all hepatocytes of the lobule. The fat vacuoles in the periacinar
hepatocytes coalesced to form larger vacuoles; and after rupture of adjacent fat-laden hepatocytes, fatty cysts
appeared. Fibrosis began in the periacinar zone associated with the ruptured fatty cysts and continued until
there was widespread bridgingperiacinar fibrosis. Islands of regeneratinghepatocyteswere frequently sequestered
within the bands of fibrous tissue. Characteristically,the hepatic and posterior mediastinal lymph nodes, lung,
and spleen contain ceroid. No lesions of hepatic encephalopathy were found in any animal. HFC is a progressive,
chronic disease of sheep, and the morphology of the hepatic lesions is similar to lipotrope-deficientforms of
nutritional cirrhosis. These findings are discussed in relation to similar nutritional deficiencies and toxicoses in
sheep.

Key words: Ceroid; cirrhosis; fatty change; liver; ruminant; sheep.

The earliest descriptions of hepatic fatty cirrhosis imals during the late winter and spring, thus causing
(HFC) demonstrated it to be a disease responsible for the initial hepatic damage. Because of drought con-
serious economic losses in livestock in a five-county ditions, the reduced range of vegetation through late
area in western Texas4Typically, morbidity varies from summer and fall creates a nutritional stress on an al-
85 to loo%, and mortality approaches 85% over an ready diseased liver, and fat mobilization with hepatic
extended 3-year period. The disease primarily affects fat accumulation continues. This is aggravated in ewes
domestic and sylvatic ruminants-chiefly, sheep, goats, due to most animals being in the last trimester of ges-
cattle, deer, and pronghorned antelope.'^^ Animals ini- tation.
tially show signs of failure to gain and maintain body Since the original description of HFC in 1932, the
weight. Signs typically associated with hepatic enceph- bulk of investigative work has concentrated on iden-
alopathy, including head pressing, depression, abnor- tifying an etiologic agent and epizootilogic factors as-
mal head positioning, and coma, are displayed in an- sociated with the disease. Cases of HFC have been
imals in the final stages of the disease. Necropsies of diagnosed in southern Texas, northeastern Mexico, and
affected sheep having clinical signs of terminal hepatic eastern New Mexico and in Texas counties surround-
disease reveal livers with varying degrees of pale yellow ing the five-county area where HFC was originally de-
discoloration and firmness believed to represent fatty As an economically important and unique
degeneration and cirrhosis, respectively. Regional disease of ruminants, it is important for veterinary
lymph nodes frequently had a similar yellow discol- pathologists to recognize the lesions of HFC and their
~ration.~ progression in order to make an accurate diagnosis.
HFC has appeared in years when there was above- The purpose of this paper is to characterize the gross
average winter rains supporting a bloom of spring veg- and microscopic lesions of HFC in sheep.
etation.’ Furthermore, in years of HFC occurrence, the
wet winters have been consistently followed by drought Materials and Methods
conditions through the summer months. Lambs born All procedures, examinations, and determinations were
in the spring following wet winters have lesions of HFC conducted on spontaneously affected sheep. Two groups of
when slaughtered 4-5 months later. Therefore, it has animals used in this study were Rambouillet ewe sheep, 2-
been suggested that a hepatotoxin is consumed by an- 6 years old, purchased off one ranch located in an HFC-
635

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636 Helman, Adams, and Bridges Vet Path01 3 2 6 , 1995

Fig. 1. Liver; sheep. The earliest macroscopic lesions are


multifocal, pale areas of fatty change (arrows) around the Fig. 3. Liver; sheep. The visceral capsular surface is al-
large blood vessels in the porta hepatis. most diffusely affected by fatty change that is represented as
homogeneous to lacy pale regions.
affected area of western Texas in Reagan County. One group
of 400 sheep was maintained on range at the ranch of pur- Tissues were fixed by immersion in 10% neutral-buffered
chase for a 7-month period from February through August. formalin, embedded in paraffin, sectionedat 6 pm, and stained
This ranch had had consistent problems with HFC over the with hematoxylin and eosin. Frozen sections of hepatic lymph
previous 20 years, such that in years of HFC occurrence 99% nodes, kidneys, lung, and liver were stained with the Oil Red
of the sheep on the ranch developed the disease. 0 method to demonstrate cellular lipids. Paraffin-embedded
The second group of 100 animals included 80 sheep with sections of liver were stained by Wilder’s silver impregnation
various stages of HFC and 20 unaffected sheep. These ani- method for reticulin and by Van Gieson’s picrofuchsin tech-
mals were brought to Texas A&M University, College Sta- nique for collagen fibers. Ceroid was demonstrated with the
tion, Texas, for study over a 1-year period beginning in March. acid-fast stain of Ziehl-Nielsen and with ultraviolet fluores-
The latter sheep were maintained on native pasture grasses cent microscopy in unstained sections. Microscopic lesions
in the spring, summer, and fall and were dewormed at 1-3- in samples taken from multiple sites in affected livers were
month intervals. During the winter, the sheep were supple- coordinated with the macroscopic lesions.
mented with prairie grass hay and grain, mostly milo maize.
The characteristics of the macroscopic lesions are based
Results
on the examination of 38 1 livers obtained by necropsy after All sheep in both groups of the study developed
HFC. The earliest gross lesions were focal to multifocal
natural death or slaughter of sheep in the first group of sheep
during the months of May, June, and August (months 4,5 , and consisted of pale, yellow, and firm subcapsular
and 7). Results were tabulated and sketches were made of areas around the porta hepatis on the visceral surface
the distribution, size, and extent of the macroscopic lesions.
of the liver (Fig. 1). The papillary process and the
Data on changes in other organs were gathered while per-
ventral border of the left lobe sometimes were involved
forming complete necropsies on 65 additional sheep, the sec-
ond group, affected with various degrees of HFC. Sheep in
to a minor extent. The lobular pattern of the liver was
the second group were necropsied at 1-month intervals accentuated throughout the organ and characterized
throughout the year-long period of study at Texas A&M. by a yellow, 1-mm central focus surrounded by nor-
mal-appearing tan to red-brown parenchyma.
Later, livers of affected sheep had moderately ex-
tensive lesions extending from the subcapsular areas
deeper into the parenchyma of the liver. The areas of
discoloration were firmer and completely surrounded
the porta hepatis extending along the boundaries of the
caudal vena cava and the attachment of the lesser
omentum (Fig. 2). The lesions located along the um-
bilical fissure and the ventral border of the left lobe
were more extensive also, having expanded toward the
porta hepatis and appearing on the parietal surface of
the liver.
In animals with more advanced HFC, there were
Fig. 2. Liver; sheep. Pale areas of fatty change (arrows) extensive lesions involving the entire parenchyma be-
have extended around the entire porta hepatis and into the tween the parietal and visceral surfaces of the left lobe
surrounding right and left lobes. (Fig. 3). The lesions were confined mostly to the retic-

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Vet Pathol326, 1995 Ovine HFC 631

Fig. 5. Liver; sheep. Earliest microscopic change in HFC


is the presence of finely vacuolated cytoplasm of periacinar
hepatocytescharacteristicof lipid accumulation.Central vein
is just out of the field of view. Hematoxylin and eosin. Bar
= 25 Fm.

vascular anastomoses and fibrous adhesions between


the hepatic capsule and the diaphragm. The afferent
lymphatics of the hepatic and caudal mediastinal lymph
nodes frequently were distended and tortuous. The per-
icardial sac commonly contained between 30 and 100
ml of clear, yellow, thin fluid. The thoracic and ab-
dominal cavities of these animals contained from 50
Fig. 4. Liver; sheep. There is prominent hypertrophy of
the margins of the liver accompanying the extensive fatty to 200 and 500 to 2,000 ml, respectively, of a similar
change and atrophy of the more central regions of the liver fluid. The mesentery commonly was very edematous.
(a). A cross-section adjacent to the hilar region of the liver The hepatic lymph nodes were enlarged and measured
showing sharp demarcation between fatty, atrophic, and hy-
pertrophied liver (b).

ular impression of the left lobe and to the omasal im-


pression of the right lobe. The caudate process was
occasionally involved, and the left lobe was atrophic
in some sheep.
In sheep with the terminal stages of the clinical dis-
ease, the surface of the liver was yellow and uniformly
pitted with small depressions. The more central regions
of both major lobes were firm and small while the right
lateral borders of the left and right lobes typically had
undergone massive hypertrophy (Fig. 4). The capsule
of the hypertrophied regions of liver was thickened and
had a lacy, gray to tan pattern. In some animals, the
entire liver was yellow and markedly atrophic. In the
atrophic central regions of the major lobes, there fre-
quently were bulging, 0.5-2.0 cm, green biliary cysts
Fig. 6. Liver; sheep. Hepatocytes with more advanced
adjacent to the caudal vena cava and porta hepatis. fatty change adjacent to the periacinar region (central vein
In more severely affected sheep, the gall bladder usu- out of view). Individual hepatocytes have one to numerous
ally was distended with dark green, viscous bile. The cytoplasmic fat vacuoles. Sinusoids are inconspicuous be-
parietal capsule of the liver often was thickened and cause of hepatocellular enlargement with fat. Hematoxylin
edematous, and in severely damaged livers there were and eosin. Bar = 25 wm.

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638 Helman, AdamIS, and Bridges Vet Pathol326, 1995

Fig. 7. Liver; sheep. Multiple fatty cysts in the periacinar Fig. 8. Liver; sheep. Portal region with hyperplastic bile
region (central vein out of field). There is early fibrosis ad- ducts, bands of fibrosis, and a regenerative hepatocytic nod-
jacent to the fatty cysts with minimal numbers of mono- ule. Hematoxylin and eosin. Bar = 100 pm.
nuclear inflammatory cells. Ceroid-laden mononuclear phag-
ocytes (see inset) are adjacent to hepatocytes and fatty cysts.
Hematoxylin and eosin. Bar = 50 pm. position of collagen occurred initially on the collapsed
fatty cysts and condensed hepatic reticulin. Fibroplasia
up to 3.5 cm. The cortex had many small, 1-2 mm, continued along a line linking adjacent central veins.
yellow-orange foci throughout. The caudal mediastinal Increased numbers of Kupffer cells were prominent in
lymph node had a similar but less pronounced ap- the periacinar regions where fibrosis was greatest. In
pearance. The thoracic and abdominal fat frequently the later stages of the disease, bands of fibrous con-
displayed serious atrophy. nective tissue with entrapped fatty cysts extended be-
Microscopically, the initial lesion was an accumu- tween central veins and portal areas, resulting in pseu-
lation of many fine, intracellular lipid droplets in the dolobulation. When fibrosis was advanced, there were
hepatocytes located in the periacinar region of the lob- often secondary hypoxic changes of the residual he-
ule (Fig. 5). The lipids stained intensely with Oil Red patic tissue in the periacinar and periportal regions
0. With progression of the disease, the lipid accu- characterized by coagulation necrosis. Secondary bil-
mulation in hepatocytes spread toward the portal areas iary hyperplasia occurred frequently in the portal areas
of the hepatic lobule and eventually affected practically and often extended along the usually indistinct bile
all of the hepatocytes. Often, mitotic figures, one to ductules adjacent to the terminal branches ofthe portal
three per 400x field, were prominent in hepatocytes veins (Fig. 8). Nodular hyperplasia of the hepatocytes
throughout the lobule. with lobular hypertrophy was noted in the right lateral
Coalescence of smaller lipid droplets to form larger borders of the right and left lobes of the liver.
lipid vacuoles was obvious within hepatocytes located Kupffer cells in and around the fibrotic areas con-
in the periacinar region, spreading peripherally in the taining fatty cysts commonly had abundant intracy-
hepatic lobules until a majority of hepatocytes were toplasmic pigment interpreted to be ceroid based on
laden with one or more large lipid vacuoles. Over time, resistance to organic solvents and yellow-brown au-
the larger intracellular vacuoles continued to increase tofluorescence under ultraviolet light (see also Fig. 7).
in size, particularly in those hepatocytes of the peria- Similar collections of pigmented mononuclear phag-
cinar region of the hepatic lobule (Fig. 6 ) . Eventually, ocytes were present in lymphatics in portal triads and
adjacent, enlarged hepatocytes ruptured, giving rise to the hepatic lymph nodes. In addition, there were in-
fatty cysts with multiple hepatocyte nuclei compressed creased numbers of mononuclear phagocytes in the
at the periphery. Such fatty cysts were most frequently cortical and medullary sinuses and lymphoid follicular
located eccentrically to or surrounding the central veins hyperplasia throughout the cortex.
(Fig. 7). In the final stages of the disease, the fibrous tissue
The first appearance of fibrosis was in the sites around became the predominant element with few normal,
the central veins where fatty cysts were most numerous discernible landmarks remaining. An occasional slump
(Fig. 7). Often, the larger fatty cysts had ruptured, re- of surviving, regenerating hepatocytes was present ad-
sulting in the release of lipids into the sinusoids. De- jacent to or surrounding a branch of a portal vein in

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Vet Path01 325, 1995 Ovine HFC 639

proximity to a fibroticportal triad. The number of fatty ventral lobe of the liver may represent a streamlining
cysts was reduced as the fibrosis became the predom- effect, which has been defined in dogs and cats.3It has
inant feature. been suggested that portal blood from the caudal mes-
The caudal mediastinal lymph nodes, spleens, and entenc and splenic veins is distributed to the dorsal
alveolar capillaries of severely affected sheep also con- lobe while blood from the cranial mesenteric vein flows
tained ceroid-laden macrophages. In the spleen, pig- into the ventral lobe. Although the exact percentage of
mented macrophages initially appeared in the central distribution is not known in ruminants, it may help
portions of the Malpighian corpuscle adjacent to the explain the differences in involvement between the
central artery but, after time, were increased in and dorsal and ventral lobes of the liver in sheep with HFC.
around the corpuscles. In advanced HFC, there was Lesions of portal hypertension are a common se-
lymphoid depletion of the spleen. No lesions of hepatic quela to the severe liver cirrhosis in sheep in the later
encephalopathy were found in any animal in the ter- stages of HFC. This is evident by the development of
minal stages of HFC. vascular anastomoses on and between the parietal he-
patic capsule and diaphragm. Distension of the afferent
lymphaticsto the hepatic and caudal mediastinal lymph
Discussion nodes plus ascites are also commonly associated with
HFC in sheep is a distinct and characteristic hepatic portal hypertension.*However, the ascites may also be
disease in which there is a progressive fatty change of exacerbated by hypoalbuminemia, which has been de-
the liver leading to cirrhosis. The development of he- scribed in sheep with HFC and would be expected in
patic lesions consistently follows a pattern with the animals with chronic liver failure.'J4
parenchyma adjacent to the major vessels and capsule HFC in sheep closely resembles experimental nu-
along the portal and umbilical fissures being affected tritional cirrhosis in laboratory r o d e n t ~ . ~The
, ~ J dis-
~
first with gradual extension into the right and left lobes ease in laboratory animals has typically been induced
of the liver. The development of hypertrophy in the by feeding a lipotrope-deficient diet in which the feed
least affected regions of liver occurs in the late stages has low levels of methionine, folic acid, and/or vitamin
of the disease and is a presumed, compensatory re- BIZ.As in HFC, the development of fatty change in
sponse to increase liver mass to meet bodily needs. nutritional cirrhosis due to lipotrope deficiency begins
All morphological hepatic lesions of HFC can be in the periacinar region and extends to involve the
related to the abnormal accumulation of intrahepa- entire lobule. Similarly, fatty cysts form around the
tocytic lipid. The gradual distension of hepatocytes central veins followed by rupture and the development
with lipid is followed by rupture of neighboring he- of fibrosis eventually progressing to cirrhosis with cer-
patocytes and formation of fatty cysts. It has been shown oid formation.
in experiments on other forms of fatty cirrhosis that Lipotropic agents are essential for the processing of
fibrosis will OCCUT only if the accumulation of lipid is intracellular lipids destined for export from the liver.12
sufficient to rupture hepatocytes and form fatty cysts.2,6 It is suggested that the synthesis and/or secretion of
As fatty cysts undergo dissolution, fibrosis develops by triglycerides in the form of very low-density lipopro-
condensation of reticulin and production of collagen tein (VLDL)is blocked with lipotrope deficiency. Stud-
by fibroblasts, and possibly myofibroblasts, stimulated ies have shown that lipotrope-deficient diets result in
by the presence of extracellular lipids, especially cho- a decrease in intracellular phosphatidyl choline (leci-
lesterol-containing lipids. thin), which is an essential component in VLDL syn-
The appearance of ceroid in hepatic lesions, regional thesis.
lymph nodes, spleen, and lung follows the rupture of The closest situation to lipotrope deficiency in ru-
fatty cysts and the onset of fibrosis. Ceroid is com- minants has been described in ovine white liver disease
monly seen in adipose tissue of animals with vitamin (WLD)I6J7In this condition, it is speculated that a soil
E deficiency and is believed to represent an abnormal cobalt deficiency results in lowered levels of vitamin
lipid product formed by the polymerization of oxi- BIZin the animal. Sheep with WLD have a diffuse and
dized, unsaturated fatty acids.5 The amount of ceroid progressive periacinar hepatocellularfatty change lead-
correlated directly with the severity and extent of fatty ing to formation of fatty cysts and accumulation of
cyst formation and rupture in sheep with HFC. For ceroid. Fibrosis along the tracts of fatty change with
the most part, ceroid is believed to be formed locally portal biliary hyperplasia also is described. Grossly,
in the liver in the regions of the fatty cysts and appears the liver of animals with WLD is diffusely pale tan to
in distant lymphoid organs and the lungs as a result of gray and becomes firm but does not have the charac-
lymphatic and/or venous transport of ceroid-laden teristic pattern of macroscopic lesion development as
mononuclear phagocytes. seen in sheep with HFC.
The earlier and more extensive involvement of the In WLD, sheep typically develop lesions and clinical

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640 Helman, Adams, and Bridges Vet Pathol 32:6, 1995

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Request reprints from Dr. R. G. Helman, Department of Pathobiology, Texas Veterinary Medical Center, Texas A&M
University, College Station, TX 778434467 (USA).

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