Photosensitivity in South Africa. V. A comparative study of the pathology of the ovine hepatogenous photosensitivity diseases, facial eczema and geeldikkop (Tribulosis ovis), with special reference to their pathogenesis

The subject of this study was the pathological and scanning electron microscopical changes in the biliary systems of sheep suffering from facial eczema or geeldikkop (Tribulosis ovis), or made photosensitive by ligation of the common bile duct. While an obliterative cholangitis is responsible for the retention of phylloerythrin in facial eczema, the occlusion of bile ducts with crystalloid material (microliths) appear to perform a similar function in geeldikkop. The similarities and differences between the 2 diseases are discussed in the light of their pathogenetic mechanisms.     

Crystal-associated cholangiopathy associated with the ingestion of Panicum spp. and other plants

Several panicoid grasses and some other plants throughout the world have been associated with a crystal-associated hepatopathy causing photosensitisation in ruminants. We have prepared sheep with reentrant bile duct cannulae and fed them on Panicum dichotomiflorum from the Waikato region. Bile from these sheep has been collected and crystals harvested by centrifugation and Percol density gradient techniques. These sheep, when slaughtered after showing elevations of serum bilirubin and enzymes indicating liver injury, had the typical cholangiohepatopathy of Panicum spp. toxicity. The crystals have now been identified as calcium salts of epismilagenin glucuronide. Likely precursor saponins of the sapogenin have been identified in Panicum dichotomiflorum. While we regard these findings as an important step forward in the understanding of these diseases, we are now investigating the factors, including known hepatotoxins, responsible for the occurrence of these hepatopathies.     

A syndrome of hepatogenous photosensitisation, resembling geeldikkop, in sheep grazing Tribulus terrestris

Two outbreaks of photosensitivity disease occurred in weaner sheep in south western New South Wales during early autumn 1982. In each instance there was a history of access to the annual herb, Tribulus terrestris and both the clinical and pathological findings were consistent with geeldikkop, a major disease in the Republic of South Africa. The prevalence rates of clinical cases were 21 and 37%, while the case fatality rates approached 70%. Clinical signs were dominated by jaundice and photosensitisation. Ochre and khaki discolouration were present in the liver and kidneys, respectively. Histopathologically, the most characteristic lesion was the presence of acicular, cholesterol-like clefts in the lumens of bile ducts and in the cytoplasm of hepatocytes and Kupffer cells. Similar structures were also evident in the lumens of nephrons in association with segmental hyperplasia of the neighbouring tubular epithelium. The possible pathogenesis of the hepatogenous photosensitisation and its resemblance to geeldikkop are discussed.     

Pathophysiologic effects of experimentally induced Fascioloides magna infection in sheep

The pathophysiologic responses of 13 sheep inoculated orally with 100 metacercariae of Fascioloides magna were monitored for 4 months after inoculation. There were no differences in weight gains between these and a number of noninoculated control sheep throughout the experiment. Complete blood cell counts showed an increase only in the absolute number of eosinophils. Serum preparations (2 times a week) from 7 inoculated and 7 noninoculated sheep did not identify any significant changes in alkaline phosphatase, aspartate aminotransferase, alanine aminotransferase, and gamma-glutamyl transferase activities. There were no changes in total bilirubin, BUN, creatinine, inorganic phosphorus, calcium, albumin, chloride, potassium, and sodium values. Four months after they were inoculated, all sheep were necropsied, and flukes were recovered. Gross lesions attributed to fluke migration were found in the liver and portal lymph nodes, diaphragm, lungs, kidneys, and spleen. Microscopically, liver lesions in inoculated sheep occurred in the portal areas, veins, and Glisson's capsule and were characterized by both active and chronic forms of inflammation. Abundant infiltrates of eosinophils and plasma cells often marked the portal areas. Endophlebitis, with or without thrombosis, was the predominate vascular lesion. The flukes recovered varied greatly in size (6.5 to 48.0-mm long) and demonstrated some sexual development, but none was sexually mature.     

Clinicopathological studies on facial eczema outbreak in sheep in Southwest Turkey

After very hot summer, 22 sheep from 5 different flocks consisting of approximately 150-200 animals each were diagnosed with facial eczema in September 2005, in southwest Turkey. Photophobia, corneal opacity, severe ulcers of the facial skin, especially localized around the eyes and mouth, and 3% mortality were the most prominent clinical symptoms. GGT levels of the animals were very high and varying between 261- 328 U/l. While the activities of ALT and total bilirubin were elevated and AST was normal in affected sheep. Total bilirubin level was higher than normal. Seven of the 22 sheep were euthanatized and necropsy was performed on all of these animals. Severe icterus, hepatomegaly, enlarged gallbladder, congestion of mesenteric vessels were the common necropsy findings. Histopathological changes of the liver included necrosis of the hepatocytes, cholangiohepatitis characterized by mononuclear inflammatory cell infiltrate in the portal area and mild to severe fibrosis around bile ducts. A diagnosis of sporidesmin toxicosis was made based on the histopathology of the livers, the elevation in liver enzymes, and the development of cutaneous lesions consistent with photosensitization and high spore counts in the ruminal contents. Surviving sheep were treated with procaine penicillin + dihidrostreptomycin sulfate, multivitamin complexes and flunixin meglumine. Additionally, zinc sulphate was also given at a dose of 6 gr per 100 lt drinking water for 28 days. All treated sheep recovered. Pasture spore counts were between 96,300- 267,500 spores/g grass.     

Lesions of spinal cord parelaphostrongylosis in sheep. Sequential changes following intramedullary larval migration

Spinal cord nematodiasis epidemiologically, clinically, and histologically consistent with Parelaphostrongylus tenuis infection was noted in two flocks of sheep. Spinal cords from two sheep with active infection and one from a partially recovered animal were studied in an effort to determine the sequence of lesions following larval invasion of the central nervous system. In the former two sheep, migration of larvae within the spinal cord induced asymmetrically irregular tracks of disrupted and necrotic tissue, primarily in white matter. Subsequently, macrophages infiltrated these regions and phagocytized the necrotic tissue, which led to cavity formation. Swelling and loss of axons, diminished myelin staining, mononuclear cell infiltration and increase in astrocytic fibers were often seen in adjacent tissue. Only occasional coiled larvae were found in these actively infected animals. Late stage lesions in the white matter in the partially recovered sheep included multiple small astrogliotic regions with diminished myelin and axonal content, and a single large multicavitary, atrophic, gliotic zone.     

The toxicity of Senecio inaequidens DC

This study was designed to confirm the toxicity of a plant implicated in an outbreak of poisoning of stock in Frankfort, Free State Province, South Africa. Cows died acutely after being introduced into a camp, where an abundant, green shrublet was noted to be heavily grazed. This plant was subsequently identified as Senecio inaequidens DC. (Asteraceae) by the South African National Biodiversity Institute (SANBI). Extraction and chemical analyses for pyrrolizidine alkaloids (PAs) in Senecio inaequidens revealed the presence of 4 different compounds, namely retrorsine and senecionine (known to be hepatotoxic) and 2 unidentified compounds. The average total PA (free base plus N-oxide) concentration in plant parts of S. inaequidens collected at Frankfort during the outbreak was 0.81%, compared with the total alkaloid content in the dried, milled S. inaequidens plant material, collected 7 weeks after the outbreak, of only 0.18%. Male Sprague-Dawley rats (n = 4), aged 8-9 weeks, were dosed per os. Each rat received a different dose of the crude Senecio inaequidens extract, ranging from 0.049 mg/g body weight (b.w.) to 0.25 mg/g b.w. No clinical signs were observed in the rat receiving the lowest dose. Rats receiving higher doses showed depression, an unsteady gait, pilo-erection and jaundice, which was particularly noticeable in the ears. Clinical chemistry evaluation revealed an increase in the activities of ALP (except Rat 4), AST and GGT in all animals. Total serum bilirubin, creatinine and urea concentrations were also elevated. All rats had low serum globulin concentrations with an A/G ratio above 1.2. Post mortem examination of the rats revealed marked hepatic lesions. Histopathologically, these changes were characterised by necrosis (variable in extent) of the centrilobular and midzonal hepatocytes (but sparing the portal hepatocytes), with extensive haemorrhage and congestion. Proliferation of the bile ducts, fibrosis and oedema were also present. Ultrastructural changes in affected rats were characterised by margination of chromatin, the presence of numerous autolysosomes in necrotic hepatocytes, intramitochondrial woolly inclusions and changes in the endoplasmic reticulum. A sheep, also dosed with the crude extract, failed to exhibit clinical signs, clinical chemistry aberrations or macroscopic lesions; however, examination of the liver of this sheep revealed histopathological and ultrastructural changes similar, though milder, to those displayed by the rats. Pyrrolizidine alkaloids were extracted from the liver and kidneys of the rats and the sheep. In the case of the sheep, retrorsine was also detected in the lungs, urine and bile.     

A comparison of parameters used to assess liver damage in sheep treated with carbon tetrachloride.

Changes in serum enzyme levels, liver histology and liver function tests have been correlated to determine the usefulness of these tests in assessing liver status. The effects of carbon tetrachloride administration on these parameters has been determined in a group of 20 sheep. Normal levels, elevated levels after injury and the effect of elapsed time after injury are reported for serum glutamic dehydrogenase, sorbitol dehydrogenase, glutamic-oxaloacetic transaminase, glutamic-pyruvic transaminase, lactate dehydrogenase, fructose-1-phosphate adlolase, alkaline phosphatase, cholesterol and proteins. Variation in the time of elevation of enzyme activities may be useful in determining the elapsed time between acute injury and serum sampling. In comparison to sheep fed an adequate diet, a diet with a restricted protein intake was associated with increased severity of histological lesions and decreased liver function.     

Isolation and typing of Clostridium oedematiens (Cl. novyi) from cases of black disease of sheep in Iran

Clostridium oedematiens types A, B and D have been isolated from liver lesions of 44 cases of black disease of sheep in Iran. The technique of isolation and identification by using fluorescent labelled antibodies is described. The isolates were typed based on their lecithinase, haemolytic, necrotic and lethal activities.     

Acute monensin toxicosis in sheep: light and electron microscopic changes

Monensin was administered orally to 3 sheep at dosages of 12 (the LD50), 16, and 24 mg/kg of body weight, respectively. Clinical signs of monensin toxicosis were observed in the sheep in 24 to 36 hours of administration. Clinical signs included CNS depression, anorexia, diarrhea, and stiffness. Increased serum creatine phosphokinase and aspartate aminotransferase activities identified possible muscle damage. Sheep were euthanatized at 54 hours after dosing; at necropsy, there were skeletal muscle hemorrhages, pale myocardium, and pulmonary edema. Ultrastructural lesions were in the liver, diaphragm, and myocardium; diaphragm and myocardium were most severely affected. Mitochondrial swelling and cristolysis, swollen sarcoplasmic reticulum, and disruption of myofibrillar architecture were prominent. These ultrastructural changes are consistent with the hypothesis that monensin causes muscle cell necrosis due to its ionophorous properties and disruption of cellular Na+:Ca2+ balance. It is proposed that this upset of normal ionic processes allows increased intracellular calcium, which directly leads to the functional and structural mitochondrial changes observed.     

Tissue and intracellular distribution of rhodanese and mercaptopyruvate sulphurtransferase in ruminants and birds

Cyanide detoxification is catalysed by two enzymes: rhodanese [thiosulphate: cyanide sulphurtransferase, E.C. 2.8.1.1], and 3-mercaptopyruvate sulphurtransferase [3-MST, EC. 2.8.1.2]. In the present work, the activity of the two enzymes in the crude extracts of different tissues and in the mitochondrial and cytosolic fractions of tissues from some ruminants (camels, cattle and sheep) and birds (chickens and pigeons) have been compared. Rhodanese activity was almost exclusively present in the mitochondrial fraction. In ruminants and chickens the highest activity of rhodanese was found in the liver, followed by the kidney. In pigeons, however, the enzyme activity was the highest in the kidneys. In camels' tissues, the rhodanese activity was significantly (P < 0.05) lower than in cattle or sheep, and the enzyme activities in the two latter species were similar. The activity of 3-MST in the crude extract of tissues from camels was similar to that in sheep, but higher than that in cattle. The enzyme activity was equally distributed between the mitochondrial and cytosolic fractions in the liver and kidneys of camels, cattle and sheep.     

A serum enzyme increasing effect of non-toxic herring meal fed to sheep

In a series of experiments it was demonstrated that highly increased activities of aspartate aminotransferase (= GOT), total lactate dehydrogenase (LDH), and α-hydroxybutyrate dehydrogenase (HBD) occurred in serum of sheep on herring meal feeding. The alanine aminotransferase (A1AT = GPT) level remained unchanged. The enzyme increase was apparently not related to the liver toxic agent dimethylnitrosamine (DMNA) occasionally occurring in lethal doses in meals produced from raw materials preserved with excesses- of nitrite. Histological changes of the liver or other tissues were never detected in experimental animals killed at a stage with very high serum enzyme values. Diets equivalent in digestible crude protein consisting of vegetable as well as of animal protein sources other than fish meal, did not give rise to elevated serum enzyme values.Electrophoretic separation of the LDH isoenzymes in serum of herring meal-fed sheep showed an increased percentage of the LDH₁ fraction, which is predominant in liver, heart, and kidney. Determination of enzyme activities in various tissues resulted in a markedly higher concentration in livers from herring meal-fed animals than in sheep fed casein at an equal protein level. It is suggested that herring meal may have a special promoting effect on the de novo synthesis of the enzymes concerned.As a practical consequence of the experiments it must be emphasized that serum determinations of these enzymes for diagnostic purposes will give a false picture of the clinical condition of sheep fed even moderate amounts of herring meal.     

Alveld-Producing Saponins: II. Toxicological Studies

The phototoxic lamb disease alveld, prevalent in South-Western Norway, is caused by ingestion of Narthecium ossifragum. Earlier studies have shown that peroral administration of large amounts of crude saponins from this plant elicits the disease. Such saponins have now been purified further by 2 different methods (A and B). Two A type preparations resulted in alveld when fed to 2 lambs. The most highly purified preparation (type B) did not cause alveld in the 2 lambs tested. Lambs vary, however, in their susceptibility to the disease. Both types of preparations led to increases in serum aspartate aminotransferase, bilirubin and 5′-nucleotidase in rats when injected intraperitoneally in amounts of 50 or 100 mg/kg body Weight. Cannulation of the bile duct showed that injected saponins reduced both the volume of bile and the amounts of bilirubin and bile acids excreted. Histological changes seen in the light microscope were, except for the most peripheral parts of the liver, hardly noticable. These observations support the view that saponins are the liver-toxic agents responsible for alveld. The possibility is discussed that the effect arises through a change in the lipid environment of carrier-mediated transport systems.     

Clinicopathological investigation of primary, uncomplicated maedi-visna virus infection.

Maedi-visna virus infection in a flock of sheep in Scotland was associated with respiratory disease, neurological disease, mastitis and lameness. The major clinical signs were dyspnoea (particularly on exercise), progressive fore- and hindlimb ataxia and balance defects, mammary induration and multilimb lameness, occasionally with enlarged carpal joints. Pathological examinations revealed lesions in the lungs, central nervous system, mammary glands and joints which were consistent with those induced by maedi-visna virus. The was no clinical or pathological evidence of concurrent sheep pulmonary adenomatosis, and pulmonary bacterial infections, when they occurred, were superimposed on the lesions due to maedi-visna virus.     

Serum liver enzyme and histopathologic changes in calves with chronic and chronic-delayed Senecio jacobaea toxicosis.

Progressive changes in serum enzyme activity and liver histologic features were monitored in calves fed tansy ragwort (Senecio jacobaea)-contaminated pellets. The experiments were designed to simulate natural intoxicant ingestion conditions in relationship to the dose and duration of exposure to the toxic plant to correlate early laboratory diagnostic changes with the natural progression of the disease, thereby facilitating early diagnosis and intervention by veterinary clinicians. Eight calves were fed tansy ragwort and 4 additional calves served as controls. In group 1, 4 calves were continuously fed dried tansy ragwort mixed in a pelleted feed at a 5% concentration by dry weight until terminal liver disease developed. Serum liver enzyme (alkaline phosphatase, glutamate dehydrogenase, and gamma-glutamyltransferase) activities were monitored at weekly intervals in these calves and in the 2 controls. In group 2, 4 calves were fed the same contaminated feed for only 60 days, with return to normal feed for the duration of the trial. Two additional calves served as controls. Their liver enzyme activities were monitored every other week in conjunction with percutaneous liver biopsies. All 8 calves fed tansy ragwort-contaminated pellets developed terminal hepatopathy in either a chronic pattern (n = 6) or a chronic-delayed pattern (n = 2), with the onset of a moribund state or sudden death at 11 to 17 weeks and 27 to 51 weeks, respectively. The calves were euthanatized when classic terminal signs of hepatic encephalopathy first became evident. The clinicopathologic patterns of chronic and chronic-delayed toxicoses were typical of over 5,000 cases of field tansy toxicosis diagnosed at the diagnostic laboratory. Serum glutamate dehydrogenase was the first enzyme to increase in most animals, with a short-term increase to peak values followed by a rapid return to normal. This enzyme change was followed by increases in alkaline phosphatase and gamma-glutamyltransferase. Serum enzyme changes preceded development of recognizable histologic lesions. Vacuolar changes in hepatocyte nuclei, biliary hyperplasia, and fibrosis sequentially developed in liver biopsy specimens from each animal, whereas megalocytosis was not a predominant feature until necropsy. On the basis of our findings, we suggest that the optimal tests for diagnosis of pyrrolizidine alkaloid intoxication should consist of liver biopsy and determination of concurrent serum liver-enzyme activities.     

Propionate loading test for liver function during experimental liver necrosis in sheep

The first objective of this work was to study the conversion of propionate to glucose by liver of the sheep during experimentally induced liver necrosis. An additional objective was to determine the most appropriate sampling time after a propionate load has been given to use glucose concentration as an aid in the diagnosis of disturbed liver function. Sodium propionate (3 mmol/kg) was injected IV into 6 healthy sheep before and after they were given carbon tetrachloride (20% CCl4 in mineral oil; 0.25 ml of CCl4/kg, orally). To differentiate the effects of liver necrosis from the effects of decrease in food intake after CCl4 administration, 5 sheep which were fasted for 2 days, but not given CCl4, were studied. Microscopically, liver necrosis was observed, as well as an increase of fatty infiltration in nonnecrotic liver tissue. After sheep were given CCl4, the plasma liver-specific enzyme activities (namely, those of iditol dehydrogenase and gamma-glutamyl-transferase) were elevated. Microscopic and enzymatic changes were not observed in fasted animals. Serum sulfobromophthalein (BSP) half-life (t1/2) was markedly increased in the sheep given CCl4 treatment (t1/2 = 22.8 +/- 11 minutes) when compared with the t1/2 before treatment (t1/2 = 2.5 +/- 0.2 minutes). The BSP t1/2 did not differ between fed and fasted sheep. The t1/2 of the IV propionate load increased significantly, from 6.9 +/- 0.4 minutes in the control sheep to 12.8 +/- 2 minutes in the CCl4-treated sheep, whereas an insignificant increase was seen after fasting (6.8 +/- 1 minutes to 8.3 +/- 1 minutes.(ABSTRACT TRUNCATED AT 250 WORDS)     

Changes in the concentrations of bile acids in the plasma of sheep with liver damage

The concentration of total plasma bile acids was measured in normal sheep and in sheep in which liver damage was induced by chronic copper poisoning, ligated bile ducts or induced ketosis. All three treatments produced a rise in total bile acid concentration in plasma which was proportional to the degree of hepatic damage seen histologically and which tended to parallel changes in activity of iditol, and glutamate dehydrogenase and aspartate amino-transferase in plasma. Plasma bile acid concentration was a more sensitive method of detecting these types of liver damage than was the measurement of total plasma bilirubin concentration, and could be used to assess alterations in liver function in sheep.     

gamma-Glutamyl transpeptidase activity in sheep serum: normal values and an evaluation of its potential for detecting liver involvement in experimental lupinosis.

A brief survey of the literature on gamma-glutamyl transpeptidase (gamma-GT) activity is included in this study. The levels of activity in the serum of normal Merino sheep (13,6-32,4 mI.U/ml) were ascertained as a preliminary to following the activity through the entire course of experimentally induced ovine lupinosis, a hepatotoxicosis caused by Phomopsis leptostromiformis (Kühn) Bubák. The response of the serum level of gamma-GT activity to the course of the disease was compared with that of glutamate oxaloacetic transaminase (GOT) and 2 liver function tests for the purpose of assessing its potential application in the study of this mycotoxicosis. Because the levels of activity of gamma-GT were more valuable for the early diagnosis of low grade acute intoxication and the detection of chronic liver involvement while those of GOT gave better information on the development of severe acute hepato-cellular damage, these 2 enzymes, considered together, were found to give the best information on the course of the toxicosis. The changes in gamma-GT activity during various stages of intoxication were also related to the histopathological lesions in the liver.     

Capillaria hepatica infection in wild brown rats (Rattus norvegicus) from the urban area of Milan, Italy

Forty-seven wild brown rats (Rattus norvegicus) collected from the urban area of Milan (Italy) were screened for Capillaria hepatica liver infection. The liver of each rat was grossly and histologically examined for the presence of C. hepatica adults, eggs and typical C. hepatica induced lesions. In 17 rats (36%) liver lesions consistent with C. hepatica infection were detected. Grossly, white-yellow nodules of 1-5 mm in diameter were present, either scattered on the liver surface or localized in a single lobe. Histologically, granulomatous liver lesions associated with eggs and/or worms were observed. The degree of gross liver involvement was moderate in most of the positive cases (71%). About 30 cases of C. hepatica infection in humans have been documented world-wide, most of which are reported in children from 1 to 5 years of age. Our results suggest that the potential transmission of C. hepatica to children in the study area should be considered an important health issue.