Isoenzyme patterns of phosphatases and esterases in Fasciola hepatica and Dicrocoelium dendriticum

A study was made of alkaline and acid phosphatase isoenzymes and non-specific esterases in homogenates of the trematodes Fasciola hepatica and Dicrocoelium dendriticum obtained from infected tissues of Capra hircus and Ovis aries using horizontal polyacrylamide gel electrophoresis.

The esterase patterns in F. hepatica and D. dendriticum were different. Homogenate of F. hepaticafrom both hosts gave four enzyme bands. For D. dendriticum, however, while homogenates of parasites from C. hircus also gave four bands, those of O. aries gave only three bands.

Acid and alkaline phosphatases in homogenates of both parasites showed three enzyme bands, but there was a host species difference betwen the enzyme patterns of specimens collected from C. hircus versus O. aries.     

The incidence of Dicrocoelium dendriticum in Emmental

A cross-sectional field study on the prevalence of Dicrocoelium dendriticum was performed in the Emmental. The study included 211 bovines, 170 equines, 20 ovines, 46 caprines and 23 rabbits (from 119 farms). In addition, laboratory routine diagnostic data obtained from 2.840 animals--all originating from the same area of investigation--were assessed in the same way. The infection extent concerning the different animal species were the following: bovines 46%, equines 12%, ovines 30%, caprines 48% and rabbits 9%. Univariate analyses of baseline epidemiological data identified no significant risk factors, with the exception of the type of stable used. Bovines kept in a modern free ranging stable had a significantly lower chance of infection with D. dendriticum than cattle in conventional tie stalls. The epidemiological data characterizing the area of investigation suggest the following procedure to reduce the problem of dicrocoeliosis: Pasturing animals of all ages should be regularly dewormed (e.g. every six week during pasture) using a compound effective against D. dendriticum. A treatment is especially indicated at the time after pasture in autumn or before housing the animals in winter. In spring, only animals having pastured the year before need to be treated prior to pasture in the new year. However, it is recommended to perform an economic analysis comparing costs of treatment versus putative costs of damage prior to the initiation of a strategic campaign: animal welfare aspects have to be considered. The laboratory routine diagnostic data showed infection extent similar to those of the cross-sectional study: bovines 60%, equines 24%, ovines 26%, caprines 31%, rabbits 32%. Atypical hosts such as dogs and cats exhibited low infection extent (3% and 1%, respectively), rather reflecting a gastro-intestinal passage of parasite eggs ingested by consumption of infected livers or by coprophagy of ruminant faeces.     

肝片吸虫对奶牛生理生化水平的影响及分析

[目的]试验选择济南地区3个奶牛场中5头感染肝片吸虫的奶牛和5头健康奶牛为试验动物,对感染肝片吸虫的奶牛生理生化指标进行了探讨。[方法]通过对奶牛体温、呼吸、心率和血液中红细胞、白细胞、血红蛋白和红细胞压积的测定来观察奶牛机体生理指标的影响;通过对血清中血糖、谷丙转氨酶和总胆红素的测定来观察肝片吸虫对奶牛血液生化指标的影响。[结果]感染肝片吸虫的奶牛体温、呼吸和心跳与对照组相比显著升高,血液红细胞总数、红细胞压积、血红蛋白降低,白细胞总数升高;血清中血糖、谷丙转氨酶和总胆红素的含量与健康组相比显著升高。[结论]肝片吸虫对奶牛的生理和生化水平都有一定程度的影响。     

An evaluation of the efficacy of netobimin against Dicrocoelium dendriticum in sheep

A trial was carried out to assess the efficacy of a nitrophenylguanidine compound, netobimin against Dicrocoelium dendriticum in naturally infected sheep. At a dose rate of 20 mg/kg bodyweight administered orally the drug was highly effective, producing a mean reduction of 98.9 per cent in the fluke burdens of treated animals compared with untreated controls. No side effects were observed in the treated sheep.     

Prevalence of Fasciola hepatica in dairy herds in England and Wales measured with an ELISA applied to bulk-tank milk

An ELISA developed to diagnose Fasciola hepatica infection in cattle by detecting serum antibodies was adapted and validated for use with samples of bulk-tank milk. The prevalence of the infection in 61 dairy herds was established by using serum antibody levels or faecal egg counts measured in a proportion of the cows in each herd. The correlation between the results of the ELISA and the herd seroprevalence was 0.83. Using a cut-off value of 27 per cent positive, the bulk-tank ELISA identified herds in which more than 25 per cent of the cows were infected with a diagnostic sensitivity of 96 per cent (95 per cent confidence interval 89 to 100 per cent) and a diagnostic specificity of 80 per cent (95 per cent confidence interval 66 to 94 per cent). By applying the ELISA to 623 herds in England and 445 herds in Wales, the prevalence of F hepatica infection in England was estimated to be 48 per cent (95 per cent confidence interval 46 to 54 per cent), and in Wales 86 per cent (95 per cent confidence interval 84 to 90 per cent).     

甘肃临夏绵羊双腔吸虫的感染与鉴定

为调查甘肃省临夏回族聚集地中绵羊双腔吸虫的自然感染情况,有效地指导防治工作,选取了具有代表性的屠宰场进行调查。首先经过形态学观察,对疑似病原进行分离,再通过PCR方法及DNA测序来鉴定。形态鉴定结果表明,在350只绵羊中共发现8只绵羊感染了双腔吸虫,分子生物学鉴定结果表明,这8只绵羊感染的双腔吸虫均为矛形双腔吸虫。     

Phase I biotransformation of albendazole in lancet fluke (Dicrocoelium dendriticum)

Dicroceliosis, a lancet fluke infection, is a frequent parasitosis of small ruminants and the anthelmintic drug albendazole (ABZ) is effective in control of this parasitosis. The aim of our project was to study the metabolism of ABZ and ABZ sulphoxide (ABZ.SO) in lancet fluke. Both invitro (subcellular fractions of fluke homogenates) and exvivo experiments (adult flukes cultivated in medium) were performed for this purpose. ABZ was metabolised invitro by lancet fluke NADPH-dependent enzymes by two oxidative steps (sulphoxidation and sulphonation). The apparent kinetic parameters of these reactions have been determined. In the exvivo experiments, only ABZ sulphoxidation was observed. The stereospecificity in ABZ sulphoxidation invitro was slight, with preferential formation of (+)-ABZ.SO enantiomer. In contrast (−)-ABZ.SO formation predominated in exvivo experiments. Sulphoreduction of ABZ.SO occurred neither invivo nor exvivo. The detection of ABZ oxidative metabolites indicates the presence of drug metabolising oxidases in lancet fluke.     

Efficacy of thiophanate and albendazole against natural infections of Dicrocoelium dentriticum, Fasciola hepatica, and gastrointestinal nematodes and cestodes in sheep

The anthelmintic efficacy of thiophanate and albendazole was compared in sheep with heavy infestations of Dicrocoelium dentriticum. The effectiveness of each drug was determined by counting the numbers of D. dentriticum in animals killed 21 days after treatment. In one group, the dose of thiophanate recommended for use against gastrointestinal (GI) nematodes (50 mg kg-1 live weight) was found to be 74.4% effective against D. dentriticum. Two tablets (each containing 76 mg of active ingredient) of albendazole per 30 kg live weight were given to a second group and the dose repeated after 1 week. Under this regime, albendazole was found to be 12.7% effective against D. dentriticum. The effect of each drug on other parasites was as follows; thiophanate had no effect against Fasciola hepatica or cestodes, while albendazole was 71.5% effective against F. hepatica and 100% effective against cestodes. Both drugs were highly effective against GI nematodes.     

An interactive map to assess the potential spread of Lymnaea truncatula and the free-living stages of Fasciola hepatica in Switzerland

The intermediate host of Fasciola hepatica is Lymnaea truncatula in Switzerland. The snail and the free-living stages of the parasite require a moderate climate and moisture for survival, reproduction, and transmission. In Switzerland, these conditions are present in many regions, resulting in a mean prevalence of bovine fasciolosis from 8.4 to 21.4%. An interactive map was created in order to demonstrate the relative risk of transmission by modelling the environmental conditions that promote the survival and reproduction of the larval stages of the parasite and the parasite's intermediate host. The map is based on temperature and rainfall data, soil conditions including ground water and forest cover in Switzerland. Extensive information on the free-living stages of F. hepatica and the intermediate host L. truncatula and how the development of these are affected by these environmental factors was used to create the interactive risk map.     

The transport of albendazole and albendazole sulphoxide in the lancet fluke (Dicrocoelium dendriticum)

Albendazole (ABZ) is one of the most important benzimidazole compounds possessing high activity against the lancet fluke, Dicrocoelium dendriticum. ABZ sulphoxide (ABZ.SO) is the main molecule present in the bloodstream of an ABZ-treated host. The aim of this study was to characterise the pattern of ex vivo uptake of ABZ and ABZ.SO by lancet flukes and the export of both anthelmintics from these parasites. Transport of these anthelmintics in both living and dead flukes was compared. The adult flukes were collected from mouflons (Ovis musimon) which had been infected naturally. Results showed that more lipophilic ABZ was imported to a higher extent than ABZ.SO, and that significantly higher concentrations of ABZ were detected within living flukes as compared to dead ones. The same pattern was revealed in the study of ABZ and ABZ.SO export from the flukes' bodies. In addition to passive diffusion, active ABZ uptake and active efflux of ABZ and ABZ.SO in D. dendriticum could be assumed.     

Partial characterization and isolation of 130 kDa antigenic protein of Dicrocoelium dendriticum adults

The study focused on characterizing and isolating Dicrocoelium dendriticum antigens or their fractions that could be used for the immunological diagnosis of dicrocoeliosis. Somatic (SoAg) and excretory-secretory antigens (ESAg) were analyzed by SDS-PAGE and their specificity was evaluated by Western blot with homologous and heterologous sera. The antigens were partially purified by chromatographic techniques of gel-filtration (Sephacryl S-300) and ion exchange (Hitrap-DEAE-Sepharose). Western blot analysis using sera of ovine infected with D. dendriticum revealed eight main antigenic polypeptides ranging from 24 to 205 kDa for SoAg and seven for ESAg with apparent molecular mass in the range of 26-205 kDa. We detected a specific parasite protein with an approximate molecular weight of 130 kDa in SDS-PAGE gels, arranged as a 450 kDa tetramer in native conditions. It also showed strong immunoreactivity by Western blot against ovine sera experimentally infected with D. dendriticum. Gel filtration chromatography (Sephacryl S-300) also showed other specific proteins, one of about 24 kDa in SoAg and another of about 42 kDa in ESAg. The elution conditions of 450 kDa protein (130 kDa monomer) by DEAE chromatography were similar to those from the somatic antigen (pH 7.2, 0.1M NaCl, in 29-34 ml fractions) and from the excretion-secretion antigen (pH 8.0, 0.1M NaCl, in 29-35 ml fractions). The suitability of 130 kDa polypeptide for D. dendriticum infection diagnosis was confirmed by Western blot using a pool of sera as well as individual serum samples from experimentally infected sheep. The sequence of amino termini of 130 kDa polypeptide from both fractions was the same and identical to that reported for a peptide from D. dendriticum described as a globin. This sequence also revealed an appreciable similarity with the amino end of globins from some phylogenetically related worms.     

Isolation and purification of Fasciola hepatica eggs

The present egg isolation method is both a rapid and simple technique for recovering large numbers of Fasciola hepatica eggs (1 X 10(7) eggs/gradient) from bovine bile. Bile from infected cattle was first passed through a 45 micron screen sieve. The F. hepatica eggs were collected from the surface of the screen by backwashing with a jet of distilled water. The resultant egg suspension was layered on a 60% to 100% (v/v) linear Percoll gradient prepared in distilled water. Centrifugation at 450 g for 20 min resulted in the formation of 2 visible bands and a pellet. The top band (density of 1.075 g ml-1) contained viscous debris and crystallized bile pigments. The second visible band (density of 1.093-1.099 g ml-1) consisted of a relatively pure population of F. hepatica eggs (greater than 93%) while the pellet contained only F. hepatica egg shells.