Isoelectric focusing of soluble proteins from Fasciola hepatica L, 1758 and Fascioloides magna B, 1875.

Isoelectric focusing was performed on the soluble proteins of whole-body and excretory-secretory products (ESP) of Fasciola hepatica and Fascioloides magna. Adult F hepatica flukes were recovered from experimentally infected sheep and ESP obtained from the flukes; portions of liver were cut and frozen at -70 C. Fascioloides magna adults were collected from naturally infected white-tailed deer and ESP obtained; portions of liver were collected from noninfected white-tailed deer. Adult flukes and their host tissues were homogenized and centrifuged; protein concentrations with their ESP were determined and adjusted to less than 2.50 mg/ml. Seven ESP samples from F hepatica and 1 from Fascioloides magna were subjected to isoelectric focusing with the 2 species of fluke and their respective host liver homogenates. After separation, gels were stained with silver and scanned on a laser densitometer. Protein banding patterns of the 2 species of flukes were dissimilar. In the pH range of 3.5 to 9.6, the body protein had approximately 30 peaks and ESP about 23 peaks in both species. Overall banding patterns of the body protein and ESP of both species were distinct from those of respective host tissues. Of the peaks reported as dominant, 3 of the body protein and 2 of ESP were shared between the 2 species. Fascioloides magna had more dominant peaks than F hepatica. This technique of soluble protein isoelectric focusing is simple and reproducible, and the 2 fluke species can easily be differentiated by this technique, as well as by morphologic characteristics.     

Efficacy of two formulations of albendazole against liver flukes in cattle.

Albendazole (10 mg/kg of body weight) was administered as a drench suspension or as a feed additive to 24 cattle with naturally acquired infections of Fasciola hepatica and Fascioloides magna. Cattle were euthanatized 16 to 30 days after treatment, and the number of viable flukes was counted. Viable F hepatica and F magna were decreased by 91.4% and 70.6% for drench administration and by 82.9% and 71.9% for the feed additive treatment, respectively. There was no significant difference between the efficacy of the 2 formulations in decreasing viable fluke numbers, compared with untreated controls.     

Potential for the spread of Fasciola hepatica in cattle in Oklahoma

In 29 central and western Oklahoma counties, 42 ranches were investigated for indigenous Fasciola hepatica infections and their suitability for the transmission of liver fluke. A 10-year retrospective study of Oklahoma cattle based on samples submitted to the Oklahoma Animal Disease Diagnostic Laboratory also was done. Indigenous fascioliasis was found in cattle in 12 counties (9 central and western counties identified in the combined field and retrospective studies and 3 additional eastern counties identified on the basis of the retrospective study). Factors essential for survival of free-living stages of F hepatica and for snail species necessary for propagation of the fluke existed in much of Oklahoma. Snails capable of serving as intermediate hosts for transmission of the fluke reportedly have been found in 41 of the 77 Oklahoma counties. Lymnaeid snails were detected in 4 counties in the field study.     

Preliminary report: immunodiagnosis of pre-type II ostertagiasis

Ostertagia ostertagi soluble antigens were prepared by gel electrophoresis and electrophoretic transfer onto nitrocellulose for enzyme-linked immunosorbent assays with serum probes. Serologic responses to L3-derived antigen of approximately 32 kDa may be unique and diagnostic for cattle harboring inhibited larvae, or pre-Type II ostertagiasis. Specificity was evaluated by comparing sera from pre-Type II cattle to sera from Type I, uninfected, Fascioloides magna infected, Fasciola hepatica infected or Cooperia oncophora infected cattle.     

Comparative efficacy of clorsulon and albendazole against Fasciola hepatica in cattle

In a dosage-confirmation trial, anthelmintic activities of clorsulon and albendazole against Fasciola hepatica were evaluated and compared. Twenty-eight cattle (8 to 12 months old) with natural F hepatica infections were randomly allotted to 4 groups of 7 cattle each: group 1, no treatment (controls); group 2, clorsulon suspension given orally at 3.5 mg/kg of body weight; group 3, clorsulon suspension given orally at 7 mg/kg; and group 4, albendazole paste given orally at 10 mg/kg. At necropsies, performed 7 and 8 days after treatment, control cattle harbored a geometric mean of 133.2 F hepatica, 16.0 of which were immature. Clorsulon administered at 3.5 mg/kg or 7 mg/kg resulted in greater than 99% removal of F hepatica, including immatures. Albendazole treatment resulted in a 76% overall reduction in F hepatica, including a 91% reduction of immatures. Fascioloides magna also were found in the cattle, but neither clorsulon nor albendazole caused significant reductions of the parasite. Adverse reactions to the 2 drugs were not observed.     

A controlled evaluation of albendazole against natural infections with Fasciola hepatica and Fascioloides magna in cattle

Albendazole administered to cattle at doses of 45, 35, 25, and 15 mg/kg of body weight reduced the number of viable Fasciola hepatica adults by 99.8%, 97.6%, 97.6%, and 95.7%, F hepatica immatures by 100%, 98.6%, 98.6% and 91.9% and Fascioloides magna by 96.5%, 99.1%, 94.0%, and 94.0%, respectively. Statistical evaluation by one-way analysis of variance, showed significant (P less than 0.05) differences in burdens of each fluke species between treated and untreated cattle, but differences in efficacy between any two of the four drug doses were not significant. Signs of toxicosis were not observed between the time of drug administration and slaughter 7 to 10 days later.     

Evaluation of clorsulon against immature Fascioloides magna in cattle and sheep

Efficacy of clorsulon was evaluated against infection with immature Fascioloides magna in 24 cattle and 12 sheep. Infections were induced by oral administration of 600 metacercariae/host. In cattle, clorsulon at dosages of 7 and 21 mg/kg of body weight was 65 and 100% effective against 8-week-old flukes, and 20 and 74% effective against 16-week-old flukes, respectively. In sheep, clorsulon at a dosage of 21 mg/kg was 92% effective against 8-week-old flukes. Significantly (P less than 0.05) more F magna were recovered from untreated sheep than from untreated cattle.     

Potential role of hares in the spread of liver fluke in the Netherlands

Hares (Lepus europeanus) sharing pasture with cattle from six locations in the Netherlands were examined for the presence of liver fluke (Fasciola hepatica) and shown to have prevalences of infection ranging from 0 to 41%. The mitochondrial haplotypes of liver flukes present in the hare populations were determined and compared with those found in cattle from a farm where triclabendazole resistance has been reported. Phylogenetic analysis indicated that the flukes present in the hares belonged to the same clades as those present in the cattle. A consideration of the life cycle of the liver fluke and the seasonal breeding pattern and ecology of hares supports the suggestion that hares may act as a refugia for liver fluke and as a vector for the spread of drug-resistant genotypes.     

Studies on the prevalence and laboratory transmission of fascioliasis in animals in the Kashmir Valley

In Kashmir, 85.1% of cattle, 51.3% of sheep and 14.8% of goats were found infected with Fasciola spp. The prevalence rate varied from 66.6 to 100.0%, 25.0 to 100% and nil to 66.0% in cattle, sheep and goats respectively in different months of the year. Fasciola gigantica was the predominant species in all animal species but sheep harboured both F. gigantica and F. hepatica. The prevalence of F. hepatica infection in sheep happens to be the first report from India. Lymnaea auricularia sensu stricto supported the development of F. gigantica under laboratory conditions. The incubation temperature affected the shedding of the cercariae. Snails maintained at 25-27 degrees C started cercarial shedding as early as day 20 post-infection (PI), whereas those maintained at 10-12 degrees C commenced it from day 64 PI. One out of three experimentally infected guinea pigs aged 1 month revealed adult flukes in the liver at necropsy on day 52 PI.     

Detection of stable diagnostic antigen from bile and feces of Fasciola hepatica infected cattle.

Diagnostic antigens in bile and feces from Fasciola hepatica infected cattle were detected and characterized by enzyme-linked immunotransfer blot (EITB) techniques. As sources of antigen, samples of bile, intestinal contents and feces were collected from five uninfected calves and from 10 calves with known Fasciola hepatica burdens. A band detected by EITB using a densitometer in the area corresponding to 26 kDa reacted with rabbit anti-fresh fluke antigen and infected cattle sera but not with fluke-negative rabbit sera, rabbit anti-Fasciola hepatica egg sera, Fascioloides magna positive or negative cattle sera. This band was not detected by Coomassie blue in sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE) gels or by Ponceau-S stained nitrocellulose strips. Band groups located at 104-66, 66-42, 42-26 and 25-16 kDa reacted inconsistently with the above sera. Sera from mice hyperimmunized with Fasciola hepatica excretory-secretory (ES) products detected only the 26 kDa band by EITB, without cross-reactivity with bands in the other molecular weight (MW) ranges. The results suggest that the 26 kDa antigen may consist of a stable component of ES products and/or tegument-related worm antigen. Diagnosis of Fasciola hepatica through detection of specific, stable antigens in feces of infected animals offers potential advantages over serum-based tests of better sample accessibility, discrimination between previous and current infections, and possible semi-quantitation of fluke burdens.     

两种片形吸虫感染家兔的试验比较

用大片形吸虫和肝片形吸虫感染家兔以便选择大片形吸虫对动物的最佳感染量,及明确肝片形吸虫和大片形吸虫的生物学和对动物宿主的致病力的差别。结果显示肝片形吸虫虫体在兔体内发育成熟的时间早于大片形吸虫,感染成活率更高,对动物的病理损害明显比感染大片形吸虫兔的病变要轻。本试验证实这两种片形吸虫除了形态学的差异外,在对动物致病力、病理损害等方面确实存在差别。     

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.     

Fascioloides magna infection causing fatal pulmonary hemorrhage in a steer

A feedlot steer died suddenly without exhibiting signs of clinical illness. Cysts containing Fascioloides magna were found in the liver and lung. Fatal pulmonary hemorrhage was associated with these flukes. Neither death nor clinical pulmonary disease has been previously attributed to infections of adult cattle by F. magna.     

Suitability of six lymnaeid snails for infection with Fasciola hepatica

The suitability of Fossaria (Bakerilymnaea) cubensis and Pseudosuccinea columella from Louisiana as intermediate hosts for Fasciola hepatica was compared to P. columella and Stagnicola elodes from Ann Arbor, MI, S. attenuata from Hidalgo, Mexico, Lymnaea gedrosiana from Iran and L. natalensis from Senegal. P. columella from LA was shown to be a more suitable host (51.3% became infected) than F. (B.) cubensis (15.2% and 26.4% of two populations became infected). The infection rate for P. columella from MI, was 50% and for L. gedrosiana was 32.5%, whereas L. natalensis, S. elodes and S. attenuata were refractory. F. (B.) cubensis and P. columella have some degree of suitability as intermediate hosts for F. hepatica under laboratory conditions, but field observations of their habitat in southern LA and characteristic management of cattle indicate that the former snail is more important as an intermediate host in this enzootic area. Tissue sections of suitable snails had few histopathological effects but physical damage caused by rediae was pronounced, mainly in the digestive gland and in the mantle.     

The prevalence and economic importance of bovine fasciolosis in Kenya--an analysis of abattoir data

A retrospective study covering a period of 10 years (1990-1999) was carried out using post mortem meat inspection records at the Veterinary Department Headquarters at Kabete to determine the prevalence and economic importance of bovine fasciolosis in Kenya. Meat inspection records from abattoirs in 38 districts distributed over seven out of the eight provinces of Kenya were examined. Prevalence of fasciolosis was calculated as the number of cattle found to be infected with Fasciola, expressed as a percentage of the total number of cattle slaughtered. Using the average weight and market price of a bovine liver, the monetary loss occasioned by condemnation of Fasciola infected livers was calculated. A survey was also carried out at Dagoretti slaughterhouse complex in Nairobi to determine the relative occurrence of F. gigantica and F. hepatica in slaughtered cattle. Cattle slaughtered at Dagoretti slaughterhouse originate from all parts of the country. A total of 5,421,188 cattle were slaughtered in the seven provinces of Kenya during the 10-year period and 427,931 (8%) of these cattle were infected with Fasciola. The region with the highest prevalence of fasciolosis was Western Province (16%) followed, in descending order, by Eastem Province (11%), Nyanza Province (9%), Rift Valley Province (8%), Central Province (6%), Nairobi Province (4%) and Coast Province (3.5%). The total economic loss incurred by the country during the 10-year period as a result of condemnation of the infected livers was approximately US$2.6 million. The total annual economic losses during this period ranged from approximately US$0.2-0.3 million. The highest total economic losses for the 10-year period were recorded in Western Province (US$0.8 million) and Central Province (US$0.7 million). A total of 1584 cattle originating from five provinces of Kenya were slaughtered at Dagoretti slaughterhouse over a period of two months of which 147 (9.3%) were infected with liver flukes. All the liver flukes obtained from the infected livers were identified as F. gigantica. It is concluded that fasciolosis is prevalent in cattle in all provinces of Kenya, that it causes great economic losses as a result of condemnation of infected livers, and that F. gigantica is the main species of liver flukes affecting cattle in Kenya. Local climatic factors, cattle trade, rustling and population numbers, and the presence of the snail intermediate hosts are probably the main factors influencing the incidence of the disease in the various regions of the country.     

Prevalence of mixed trematode infections in an abattoir receiving cattle from northern Portugal and north-west Spain

Between February 2007 and January 2009, a total of 776 slaughtered animals were examined for the presence of Trematoda in the liver, gall bladder, oesophagus and stomach (rumen and reticulum). Data collected were analysed by the age and breed of the animals. The percentage of cattle from which Trematoda were found was 38 per cent (95 per cent confidence interval [CI] 35 to 41 per cent); 28 per cent (95 per cent CI 25 to 31 per cent) had Fasciola hepatica, 12 per cent (95 per cent CI 10 to 14 per cent) Calicophoron daubneyi and 6 per cent (95 per cent CI 4 to 8 per cent) Dicrocoelium species. A significantly high prevalence of fasciolosis and dicrocoeliosis was observed in cattle over 10 years of age. Autochthonous Rubia Gallega cattle had the highest prevalence of fasciolosis and crossbred cattle had the highest prevalence of dicrocoeliosis (P<0.05). Twenty per cent (95 per cent CI 15 to 25 per cent) of the cattle positive for Fasciola also had Calicophoron species; 10 per cent (95 per cent CI 6 to 14 per cent) also had small liver flukes (Dicrocoelium species).     

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。     

Identification of the Egyptian species of Fasciola.

Reports on the species of Fasciola present in the Nile Delta, Egypt, appear controversial. Some authors reported the presence of both Fasciola gigantica and Fasciola hepatica, others reported F. gigantica only and mentioned that F. hepatica was found only in imported animals.This study was an attempt to identify the species of Fasciola flukes collected from locally bred animals. Morphologic, morphoanatomic, morphometric, and chemotaxonomic criteria of the fluke isolates were studied. Speciation based on morphologic and morphometric data was not decisive due to overlap in the values of most measurements. Morphoanatomic data proved the presence of both the species, and isoelectric focusing (IEF) of fluke soluble protein confirmed the presence of both F. gigantica and F. hepatica in Egypt.     

Perilobular fibrosis in chronic bovine fascioliasis

Summary The character, extent, and localization of perilobular fibrosis were studied in cattle experimentally infected with Fasciola hepatica. Such fibrosis is indirectly due to the liver flukes. The regionally localized fibrosis results from cholestasis due to transient occlusion of branches of the biliary tree. In perilobular fibrosis it is possible to distinguish a florid, an intermediate, and an uncomplicated form. The florid phase is marked by proliferation of the biliary epithelium and is followed by infiltration of inflammatory cells.