Studies on the pathology of dicrocoeliasis and fascioliasis in the goat. 3. The histochemistry of mast cells and globule leucocytes

Comparative studies were made on the structure and histochemistry of mast cells and globule leucocytes in dicrocoeliasis and fascioliasis of the goat. Investigations showed that D. dendriticum and F. hepatica infections in the goat produce an increase in the number of hepatic mast cells and frequent occurrence of globule leucocytes in the epithelium of the bile ducts. The only difference between these infections was that globule leucocytes were more frequent in small bile ducts in dicrocoeliasis than in fascioliasis.Histochemical methods showed the presence of sulphomucins in the granules of mast cells, while both neutral mucins and carboxymucins were revealed in the globules of globule leucocytes. The acidity of the mucosubstances could not be shown to result from hyaluronic acid, chondroitin-4 and -6-sulphates, or sialic acid. The globules of globule leucocytes also contained histochemically identifiable highly basic proteins.Keyword: Dicrocoelium dendriticum, Fasciola hepatica, liver, bile ducts, mast cell, globule leucocyte     

Studies on the pathology of dicrocoeliasis and fascioliasis in the goat. II. The histochemistry of bile-duct mucosubstances

Histochemical studies were made on the characteristics of carbohydrate-rich compounds in the bile-duct mucosa of goats infected either with Dicrocoelium dendriticum or with Fasciola hepatica. In a previous study the author showed that a chronic infection with these parasites produces glandular hyperplasia in the walls of bile ducts. This study revealed a definitely increased secretion of qualitatively normal mucosubstances by the hyperplastic epithelial cells.Epithelial mucosubstances in the bile-duct walls were characterized by the presence of periodate-reactive hydroxy groups and both carboxyl and sulphate radicals. The intensity of the PAS reaction was moderate or strong and not definitely inhibited by the acidic or acetyl radicals. Specific enzymatic digestions did not reveal periodate-reactivity produced by glycogen or basophilia due to hyaluronic acid or chondroitin-4 and -6-sulphates in the epithelial cells. On the other hand, sialidase digestion showed that the main part of the carboxymucins was sialidase-labile sialomucins. Their susceptibility towards sialidase was not definitely increased after deacetylation. The single and sequential histochemical methods showed a more intense staining reaction for sulphate radicals in the mucosubstances of the deep glandular and goblet cells than in those of the superficial glands, which were characterized by high concentration of carboxyl groups.Keyword: Dicrocoelium dendriticum, Fasciola hepatica, liver, bile ducts, mucosubstances     

Liver fluke infestation in New World camelids. Parasitology, pathology, clinical findings and therapy]

In Llamas and Alpacas infestation with Fasciola hepatica or Dicrocoelium dendriticum can cause liver damage, sometimes even with lethal outcome. Once infected South American Camelids (SACs) react more sensitively to these parasites than other domestic ruminants. We report here on the pathology, parasitology, clinics and therapy of this disease. Concerning Dicrocoelium dendriticum we describe own clinical results and therapeutic outcome in addition to the pathological investigation. According to anatomic corrosion casts, the bile ducts of SACs show more similarity with the equine bile system than with the bile system of domestic ruminants.     

Anatomicopathological study of vascular and biliary systems using cast samples of Fasciola-infected bovine livers

In 117 livers with fascioliasis, this study was focused on the number of Fasciola, the number and intrahepatic localization of affected hepatic ducts and bile ducts, and the degree of fibrosis in the hepatic segments and bile ducts. The degree of pathological changes in bile ducts caused by fascioliasis was classified into five levels. The site of Fasciola habitation was most often the hepatic ducts of the porta hepatis: it was the left hepatic duct in 101 livers and the right hepatic duct in 88 livers. Casts were prepared by infusing synthetic resin into the hepatic arterial, portal, hepatic venous and biliary systems of 15 bovine livers with fascioliasis and then examined. In the left lobe, quadrate lobe, and caudate process where atrophic fibrosis was noted, the bile ducts became rod-shaped by losing branches, and the samples resembled dead branches of liver. Portal branches were thinned or completely terminated with marked fibrosis. Fine and irregular newly formed bile ducts not parallel with portal branches were observed in livers with markedly chronic fascioliasis. Distal portal branches in the right lobe, caudate lobe, and papillary process showed hypertrophic proliferative changes. The arterial system was generally well developed in thickened walls of bile ducts and formed vascular beds, and surrounded the bile ducts as tubes. In livers with severe fibrosis, capillaries were markedly developed and resembled glass cotton.     

On the ultrastructure of epithelial cells in bile ducts of cattle chronically infected with fasciola hepatica

This paper describes briefly the normal ultrastructure of bile-duct epithelial cells in cattle and the main ultrastructural alterations produced in them by chronic infection with Fasciola hepatica. The studies showed only inconsiderable abnormalities in the ultrastructure of the epithelium in infections of a low degree as also during the recovery phase of the disease. On the other hand, in heavily infected livers pronounced ultrastructural alterations were seen in the surface epithelium of the main intrahepatic bile ducts invaded by the flukes. In such areas the intercellular spaces were filled with dense homogeneous material, and intact desmosomes and junctional complexes were not regularly seen, which indicates that foreign material may leak into the bile. The extracellular material was probably composed of albumin, and the significance of the present results regarding the etiology of hypoalbuminaemia during chronic fascioliasis is discussed.     

Liver flukes promote cholelithiasis in sheep

The main objective of this study was to investigate whether cholelithiasis in sheep is related to parasitism or other commonly observed disorders such as liver abscesses. Additionally, the features of the observed biliary calculi are described. The livers of 254 randomly selected clinically healthy adult dairy sheep were used. All visible concretions in the bile were considered as stones. Based on the macroscopical examination, 60 livers were normal, 40 were parasitized with Fasciola hepatica, 42 were parasitized with Dicrocoelium dendriticum, 28 were parasitized with both D. dendriticum and F. hepatica, 40 livers had abscesses and 44 had hydatid cysts. Biliary calculi were detected in 40 livers. Twenty livers had pigment stones and 20 livers had cholesterol stones. The percentage of cholelithiasis was significantly higher in livers parasitized with flukes compared with the others (P<0.05) and the parasitoses with F. hepatica and D. dendriticum were proven to be significant risk factors (P<0.05) for the presence of cholelithiasis in sheep.     

Cellular proliferation of bile ductules and gamma-glutamyl transpeptidase in livers and sera of young cattle following a single infection with Fasciola hepatica

During the course of experimental single infection with Fasciola hepatica in calves, the origin of the increased concentration of gamma-glutamyl transpeptidase (gamma-GT) found to be present in the serum during the biliary stage of the disease, was studied. The enzyme originates from the liver after flukes have caused intermittent and selective cholestasis in the bile-duct system. The enzyme which normally is present in the bile ducts can under these circumstances enter the surrounding interstitial tissues through broken-up junctions of the bile drainage system or be added to these from proliferating bile-ductule epithelium. The multiplication of bile-ductule cells can be looked upon as a neogenesis of cholangioles, which becomes essential when newly formed perilobular connective tissues prevent restoration of the anatomical connections of the bile capillaries with the biliary tree. The connective tissue growth is induced by bile fluid. An initially regional perilobular fibrosis is eventually distributed diffusely through the liver by migration of flukes. The continued and extending fibrosis is due to the constant recurrence of intermittent and selective cholestasis. The transient occlusions of branches of the bile ducts are very likely to be attributable to persistent cholangitis. The growth of connective tissues has a considerable effect on the gain in weight of the liver.     

Experimentally induced Fasciola hepatica infection in white-tailed deer. II. Pathological features.

Six white-tailed deer (Odocoileus virginianus) and six sheep were inoculated with metacercariae of Fasciola hepatica. Two animals of each species were given 100, 500 or 2500 metacercariae. One animal in each inocluated group was killed and examined at six weeks postinoculation and the remainder at 15 weeks postinoculation. At six weeks postinoculation the parietal surface of the livers from inoculated deer was covered with gray fibrous plaques and rust colored patches. Fibroplasia with mononuclear cell infiltration characterized Glisson's capsule on the parietal surface. Granulomas were found in the hepatic parenchyma and on the dorsal surface of the lung. Fresh and healing tracks were occasionally found in the liver. In the sheep fibrinous exudate and numerous subcapsular tracks were found on both surfaces of the liver. Inflammatory changes in portal areas and numerous fresh and healing tracks in the hepatic parenchyma were prominent features. At 15 weeks postinoculation inflammatory changes in Glisson's capsule of inoculated deer were less marked than at six weeks but portal fibrosis and hyperplasia of bile duct epithelium were more advanced. A zone of hemorrhage surrounded ducts that contained mature F. hepatica in one deer. The livers from the sheep were rough, pitted and covered with fibrous tags and adhesions to the diaphragm and greater omentum were common. Hemorrhagic tracks were common in the sheep given 500 and 2500 metacercariae. Portal fibrosis and hyperplasia of bile duct epithelium were seen in the sheep (100 metacercariae) that harbored mature F. hepatica.     

A cross-sectional coprological survey of liver flukes in cattle and sheep from an area of the southern Italian Apennines

A cross-sectional coprological survey of liver flukes (Fasciola hepatica and Dicrocoelium dendriticum) was conducted on 81 bovine farms and 197 ovine farms with animals pasturing in an area (3971 km(2)) of the southern Italian Apennines. The farms were selected to be uniformly distributed throughout the study area using geographical information system (GIS) software. Between June 1999 and March 2000, faecal samples were collected from 975 cattle and 3940 sheep and examined using a modified McMaster technique. The results were subjected to statistical analysis and point distribution maps (PDMs) were drawn by GIS.Cattle of 9 of the 81 (11.1%) farms were positive for F. hepatica and of 43 (53.1%) for D. dendriticum. Sheep of 8 of the 197 (4.1%) farms were positive for F. hepatica and of 133 (67.5%) for D. dendriticum. Co-infection was found in cattle of 2 (2.5%) farms, and in sheep of 8 (4.1%) farms.The findings of the present survey show that D. dendriticum was the predominant liver fluke found in cattle and sheep with respect to egg count numbers for both farms and animals. In addition, the general trends of the PDMs show that D. dendriticum was widely and homogeneously spread throughout the study area, whereas F. hepatica was present only in a few concentrated zones of the study area that had both positive bovine and positive ovine farms.     

Prevalence of Fasciola hepatica and Dicrocoelium dendriticum in the cow: inspection in an east Switzerland abattoir

On the basis of meat inspection in an abattoir in the eastern part of Switzerland, the incidence of liverflukes in cattle was recorded weekly during one year. The prevalence was compared with surveys from the seventies and nineties. The average incidence of Dicrocoelium dendriticum infection did not change. Although the prevalence of Fasciola hepatica apparently slightly decreased, an actual decrease of liverflukes could not be demonstrated.     

Percutaneous ultrasound-guided cholecystocentesis in cows.

A method was developed for percutaneous ultrasound-guided cholecystocentesis in cattle. The procedure was performed on the right side in the 9th, 10th, or 11th intercostal space of 30 cows. Of the 30 cows, 20 were slaughtered 24 hours after cholecystocentesis and the remaining 10 cows were slaughtered after a 10-day observation period. Changes in the peritoneum and gallbladder wall, observed at slaughter, were minimal. During the 10-day observation period, general behavior, attitude, and appetite of the 10 cows were normal. A transient, slight increase in rectal temperature was observed in 6 cows at 4, 5, or 8 days after cholecystocentesis. Total and differential WBC counts and total protein and fibrinogen concentrations, determined daily, were all within normal ranges. Bile samples from 20 cows were examined microscopically and biochemically. Fasciola hepatica and Dicrocoelium dendriticum eggs were observed in bile from 7 and 12 cows, respectively. Fecal examination revealed F hepatica eggs in 4 cows; D dendriticum eggs were not identified in any of the fecal samples. In 1 cow, F hepatica eggs were observed in the feces, but not in the bile. Bile acids concentration in bile varied from 12.5 to 68.5 mmol/L (mean +/- SD, 45.3 +/- 3.05 mmol/L) and in serum from 3.8 to 281.0 mumol/L (41.6 +/- 17.24 mumol/L). Negative correlation was obtained between bile acids concentration in bile and that in serum (r = -0.60, P less than 0.01). It was concluded that percutaneous ultrasound-guided cholecystocentesis in cows is a safe procedure and that microscopic and biochemical examinations of obtained bile can be useful diagnostic aids.     

On the Ultrastructure of Mast Cells and Globule Leucocytes in the Common Bile Duct of Cattle Chronically Infected With Fasciola Hepatica

Mast cells and globule leucocytes in the walls of the common bile ducts of seven uninfected slaughtered cows and of five cows chronically infected with Fasciola hepatica were studied by electron microscopy. The mast cells displayed numerous small intracytoplasmic granules, the matrixes of which were either strongly electron-opaque and homogeneous or less dense with a fine-granular ultrastructure. Unattached ribosomes were abundant in the cytoplasm of the cells. Many mast cells in the walls of chronically infected bile ducts were normal-looking while the cells in the subepithelial tissues often showed striking irregularities in the ultrastructure of the granules. Globule leucocytes were most numerous in the surface epithelium and were not connected by desmosomes to the epithelial cells. The nuclei of the globule leucocytes were similar to those in the mast cells except that indentations caused by the globules were more usually seen. The globules were considerably larger than the mast cell granules but bounded by similar agranular capsules. The matrix of most globules was dense and homogeneous while the other globules showed dense reticulate or lamellate structures. The relationship between the mast cell and globule leucocyte in the liver is briefly discussed with reference to recent reports on these cells in the intestinal wall.     

Globule leukocyte and mast cell in bile ducts of cattle naturally infected with liver flukes

In n previous report dealing with the pathology of bovine fascio-liasis the author described an unknown cell type in the epithelium of bile ducts. The histological and histochemical investigations published in this paper suggest that the cell may be considered a globule leukocyte. Globule leukocytes are rare in uninfected livers but are occurring in abundance in main bile ducts of cattle with spontaneous fascioliasis and also in small perilobular ducts in dicrocoeliasis. Liver fluke infection causes an increase in the population of subepithelial mast cells. Mast cell and globule leukocyte present similarities In their cytochemical properties. However, at low pH toluldinc blue shows a stronger but Alcian blue a weaker affinity for mast cells than for globule leukocytes.     

肝片吸虫感染致斑纹角马死亡案例调查

首次报道了国内圈养斑纹角马肝片吸虫病确诊病例。诊断依据：（1）提示性临床症状：贫、消瘦、营养不良;（2）尸体剖检特征性病理变化：胆管显著扩张和胆管内挤排出大量肝片吸虫成虫的虫体;（3）肝片吸虫感染强度测定：运用麦克马斯特法测定了病死角马肝片吸虫感染强度为1 600个/g;（4）中间宿主调查：角马的饲养区内发现大量椎实螺,椎实螺体内含有大量的肝片吸虫尾蚴;（5）组织病理学检查：细支气管和肺泡腔内见有童虫的虫体结构。首次证实了角马肝片吸虫感染时,童虫在移行过程中可误移至肺,导致肺的病理变化。该病的发生与环境潮湿多水、环境中存在大量椎实螺而未被重视有关。肝片吸虫宿主范围广,应重视易感动物的生活环境,定期粪检确定感染率和感染强度,以及选择有效的药物防治,这是科学预防该病的关键。     

Abattoir condemnation due to parasitic infections and its economic implications in the region of Trikala,Greece

The prevalence of parasitic infections responsible for the condemnation of carcasses and viscera during meat inspection, and their economic implication, was estimated in a year long abattoir survey of 10 277 slaughtered farm animals in the region of Trikala, Greece. The organs examined for the presence of parasitic lesions during meat inspection were: liver and lungs of all animals, rumen of cattle, small intestine of lambs and kids, and muscles of cattle and swine. The parasitic lesions observed in the lungs of cattle, sheep and goats were caused only by hydatid cysts. No hydatid cysts were observed in the lungs of swine. The parasitic lesions observed in the liver of cattle, sheep and goats were as a result of hydatid cysts and flukes of Fasciola hepatica and Dicrocoelium dendriticum, while those of swine were due to milk spots only. Moniezia sp. proglottids were found in the small intestine of lambs only. The prevalence of parasites responsible for the condemnation of marketable organs was low (0.26%). Parasites were responsible for 22% of the total of condemned organs, and their annual cost was 99, 00 GDR (approximately 292 Euros). The parasites most contributing to marketable organ condemnation were hydatid cysts (26%) and D. dendriticum flukes (26%).     

Fasciola hepatica: comparison of flukes from Korea and the United States by isoelectric focusing banding patterns of whole-body protein.

Comparisons were made between the flukes from Chonnam, Korea and Oregon, USA by isoelectric focusing (IEF) of whole-body protein. Adult Fasciola hepatica were recovered from bile ducts of Korean native cattle. Whole-body protein of the flukes was subjected to IEF, and the banding patterns of the fluke protein were compared with those of North American F. hepatica recovered from experimentally infected calves. The overall banding pattern of F. hepatica from Korea was essentially identical to that of F. hepatica from the United States. These results provide further support for the usefulness of this technique in differentiating Fasciola species in other geographical areas.     

PRENATAL INFECTION WITH FASCIOLA HEPATICA IN CALVES

The livers from 16,776 calves 1 to 3 weeks of age slaughtered at Shepparton abattoirs were examined for gross evidence of infection with Fasciola hepatica. Those livers with thickened bile ducts were examined for the presence of liver fluke by slicing. One hundred and eight F. hepatica were collected from 84 of these livers, giving a minimum prevalence rate of prenatal infection of calves of 0.5%. These results are discussed in context with reports from other countries.     

Light and scanning electron microscopic studies of the extrahepatic bile duct of sheep with experimentally induced Fasciola hepatica infection.

Changes in the common bile duct that contained adult Fasciola hepatica of sheep were evaluated by light (LM) and scanning electron microscopy (SEM). Nine ewes were inoculated with F hepatica metacercariae and necropsied 18 weeks after inoculation. The proximal portion of the common bile duct (CBD) that contained adult flukes was recovered and examined by LM and SEM. The CBD from 2 noninoculated ewes were used for control. On gross examination, CBD were markedly large because of the adult flukes, which were free in the lumen of the ducts. Extensive hemorrhage was not found either in intrahepatic or in extrahepatic bile ducts of any sheep. Histologic examination revealed changes, such as villous hyperplasia and hypertrophy of the epithelium; cell infiltration, predominantly with eosinophils or macrophages; and arterial intimal proliferation. By SEM, the epithelial surface of the CBD appeared intact. Villous hyperplasia and hypertrophy of the epithelium observed by LM was clearly seen by SEM. Epithelial damage, seen as small areas of denuded surface by LM and SEM, was confined to a few areas of the mucosa. Lack of extensive hemorrhage and confined epithelial damage were evaluated relative to the mode of feeding of adult flukes.     

FASCIOLIASIS IN DAIRY COWS ON IRRIGATED PASTURE

The prevalence and severity of fascioliasis was studied in 87 dairy cows from 36 irrigated farms in Victoria. Evidence of Fasciola hepatica infection was found in 85 (98%) cows representing all but one of the herds. The mean fluke burden (7 ± 6) in cows treated with anthelmintics at drying off was lower (p < 0.05) than in untreated cows (24 ± 7). Haemoglobin levels in untreated cows (9.6 ± 0.4) were higher (p < 0.05) than values obtained from treated animals (8.2 ± 0.3). Severity of bile duct wall thickness and hepatic fibrosis was similar in both groups. None of the measured parameters was affected by the age of the cow. Although fascioliasis was prevalent in these cattle the level of infection was low.