Correlation between egg-shedding and uterus development in Fasciola hepatica human and animal isolates: applied implications

The emission of Fasciola hepatica eggs in faeces is usually subject to oscillations along time in animals as well as humans. Thus, looking for alternative biological markers reflecting eggs shed per gram of faeces (epg) with lower oscillations may be useful. This study analyzes the possible relationship between liver-fluke uterus area and epg. Uterus area (UA) development of adult F. hepatica obtained at different days post infection (dpi) in a Wistar rat model with isolates obtained from cattle, sheep, pigs and humans from the endemic human fascioliasis zone of the Northern Bolivian Altiplano was analyzed and compared with the number of eggs shed per gram of faeces as obtained through the Kato-Katz technique. The morphometric study of the UA of liver flukes was carried out using image analysis software. The multiple regression model shows that UA is dependent on dpi and isolate. The evolution of UA vs dpi followed a damped model. This work shows a positive relationship between liver-fluke UA and egg production. The complete absence of eggs in the uteri of some parasite individuals at 300 dpi was observed, which corresponds to the cessation of egg shedding in the advanced chronic stage. The results obtained suggest the necessity to characterize the isolates employed with regard to geographical as well as host origin in fascioliasis studies in which egg production is used as a biological tag.     

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。     

Control of fascioliasis in heifers in Czechoslovakia 1970-1985

A programme for the control of fascioliasis caused by F. hepatica in cattle was launched in 1970 and continued until 1985. The animals were subjected to helminth-control treatment and preventive measures were taken in the herds. The anti-fasciolics Bilevon M and to some extent also Bromsalan pulv. were administered by the veterinary service on the basis of state subsidies in 1973 to 1979. The preparation Helmisan (Spofa) has been in use since 1980; its production is subsidized by the agricultural enterprises. In the period from 1972 ot 1979, up to 213 thousand helminth-control treatments (both therapeutic and preventive) were performed every year in the cattle. All treatments reduced the occurrence of Fasciola hepatica and pathological changes in the liver of the killed cattle (from 5.8% recorded in 1971 to 0.9 in 1980, on an national average). As a result, 29 districts are free from fascioliasis although the disease originally occurred in all districts of the Czech Socialist Republic. After ten years of purposeful use of antifasciolics, the findings were free from living flukes and the cattle only showed pathological changes after the action of F. hepatica. Since 1980 the extensity of the findings of F. hepatica in slaughtered cattle has ranged within one percent, though living flukes have recently occurred again in the liver of certain animals. In the enzootically affected regions and districts of the Czech Socialist Republic the fascioliasis control programme succeeded in reducing the number of invaded cattle herds.     

Studies on the pathology of dicrocoeliasis and fascioliasis in the goat. I. The histopathology of the liver and bile ducts

Comparative studies were made on the histopathology of the liver and bile ducts in dicrocoeliasis and fascioliasis of the goat. Investigations revealed certain similarities in the nature of the pathological phenomena produced by D. dendriticum and F. hepatica. However, a feature peculiar to fascioliasis was the migratory tracks of immature flukes in the liver parenchyma. Further, the lesions produced by fascioliasis were distinctly more pronounced than those present in dicrocoeliasis.In both parasitic diseases the bile-duct walls showed glandular hyperplasia, increased frequency of goblet cells and globule leucocytes, and extensive fibrosis in the outer layers. The desquamation produced by Dicrocoelium remained superficial whereas Fasciola was capable of causing deep erosions in the walls of the main bile ducts. The bile-duct walls did not become calcified. Increased fibrosis was present in the interlobular areas, too, but only fascioliasis produced extensive disorganisation of the hepatic architecture, due to the scars originating from repaired migratory tracks. The liver parenchymal cells were degenerated, showed decreased glycogen but were increasingly infiltrated by neutral fats.Keyword: Dicrocoelium dendriticum, Fasciola hepatica, liver, bile ducts     

Natural Fasciola hepatica infection in nutria (Myocastor coypus) in Uruguay

Fascioliasis, the zoonotic disease caused by the trematode Fasciola hepatica, is expanding worldwide, with a 17 million people at risk. Rodents, often recognized as a major source of zoonotic diseases, are affected by F. hepatica, with some species playing important roles in the disease epidemiology. The case reported here in a nutria or kiyá (Myocastor coypus) is the first documented case of F. hepatica in this species in Uruguay. Parasitic burden and total egg production detected are markedly higher than reported previously for this species, confirming its potential role as an effective reservoir and disseminator of liver flukes. Although further research is needed, nutria should be considered when designing effective control programs for fascioliasis.     

Myocastor coypus as a reservoir host of Fasciola hepatica in France

To clarify the role of the nutria Myocastor coypus in the epidemiology of domestic fasciolosis in Loire-Atlantique (department of western France), 438 nutrias were trapped in 9 humid areas of the department and 304 nutrias were trapped in 3 farms where Fasciola hepatica was present; all animals were necropsied. Liver flukes were found in 160 nutrias: 38 nutrias randomly taken in the department (8.7%) and 122 trapped in fasciolosis areas (40.1%). The average parasitic burden was 5.7 flukes per nutria. Sixty-five percent of the liver flukes measured more than 18 mm (size of sexual maturity). The coproscopic examinations carried out on 144 infected nutrias showed that 90% of the infected nutrias shed fluke eggs. The hatching rate was 39.6%. Two groups of 100 Lymnaea truncatula snails, originating from 2 different populations, were exposed to F. hepatica miracidiae hatched from eggs collected from infected nutrias. The prevalence of the infection was 74% and 58.6% in the 2 groups of snails. The average redial burden was 6.2 rediae per snail. The total number of metacercariae was 72.4 metacercariae per snail producing cercariae. Two groups of 5 sheep were orally infected by 150 metacercariae of nutria or sheep origin, respectively. The installation rates of F. hepatica in sheep were respectively 31.6% and 29.6% for the two groups. Specific antibody kinetics of sheep were similar whether the metacercariae were of nutria or sheep origin. M. coypus allows the complete development of F. hepatica and releases parasitic elements that are infective for domestic ruminants. Because of its eco-ethologic characteristics, the nutria could be a potential wild reservoir of F. hepatica in France.     

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: Histology of the testis in egg-producing adults of several laboratory-maintained isolates of flukes grown to maturity in cattle and sheep and in flukes from naturally infected hosts

A total of 8 calves approximately 6 months old and 22 lambs of similar age were infected with metacercariae of Fasciola hepatica of various laboratory-maintained isolates including: Cullompton (sensitive to triclabendazole) and Sligo, Oberon and Leon (reported as resistant to triclabendazole). Ten to 16 weeks after infection, flukes were harvested from these experimental animals and the histology of the testis tissue was examined in a representative sample of flukes from each population. Adult wild-type flukes were also collected from 5 chronically infected cattle and 7 chronically infected sheep identified at post-mortem inspection. The testis tissue of these flukes was compared with that of the various laboratory-maintained isolates. Whilst the testes of the wild-type, Oberon and Leon flukes displayed all the usual cell types associated with spermatogenesis in Fasciola hepatica (spermatogonia, spermatocytes, spermatids and mature sperm), the Cullompton flukes from both cattle and sheep showed arrested spermatogenesis, with no stages later than primary spermatocytes represented in the testis profiles. The presence of numerous eosinophilic apoptotic bodies and nuclear fragments suggested that meiotic division was anomalous and incomplete. In contrast to the wild-type flukes, no mature spermatozoa were present in the testes or amongst the shelled eggs in the uterus. A high proportion of the eggs collected from these flukes hatched to release normal-appearing miracidia after an appropriate incubation period, as indeed was the case with all isolates examined and the wild-type flukes. It is concluded that the eggs of Cullompton flukes are capable of development without fertilization, i.e. are parthenogenetic. The implications of this for rapid evolution of resistant clones following an anthelmintic selection event are discussed. Amongst the Sligo flukes examined, two subtypes were recognised, namely, those flukes with all stages of spermatogenesis and mature spermatozoa present in the testes (type 1), and those flukes with all stages of spermatogenesis up to spermatids present, but no maturing spermatozoa in the testes (type 2). Each sheep infected with the Sligo isolate had both type 1 (approximately 60%) and type 2 (approximately 40%) flukes present in the population. Spermatozoa were found amongst the eggs in the uterus in 64% of flukes and this did not necessarily reflect the occurrence of spermatozoa in the testis profiles of particular flukes, suggesting that cross-fertilization had occurred. The apparent disruption of meiosis in the spermatocytes of the Cullompton flukes is consistent with reports that Cullompton flukes are triploid (3n=30), whereas the Sligo and wild-type flukes are diploid (2n=20). In the Sligo flukes the populations are apparently genetically heterogenous, with a proportion of the flukes unable to produce fully formed spermatozoa perhaps because of a failure in spermiogenesis involving elongation of the nucleus during morphogenesis.     

The ability of diamphenethide to control immature Fasciola hepatica in sheep at a lower than standard dose level

Groups of eight Welsh Mountain sheep were dosed with diamphenethide at the rate of 70 mg/kg bodyweight at either one, four, six or eight weeks after artificial infection with approximately 300 Fasciola hepatica metacercariae. Comparisons were made with similarly infected but undosed sheep and with sheep which were neither infected nor dosed. The good clearance of flukes up to six weeks of age (above 97 per cent on pooled data) was reflected in the plasma concentrations of the accepted liver damage marker enzymes glutamate dehydrogenase and gamma-glutamyl transpeptidase. Highly significant correlations were demonstrated between the numbers of flukes recovered, the plasma levels of these enzymes and haemoglobin and plasma albumin values. At 70 mg/kg, diamphenethide was shown to be able to control F hepatica populations of up to six weeks of age. The systematic use of diamphenethide at this dose level at intervals of up to six weeks during the period of metacercarial challenge should prevent ovine fascioliasis.     

Emergence of fasciolosis in cattle in East Anglia

Liver fluke (Fasciola hepatica) infection caused weight loss, diarrhoea, decreased milk yield and occasionally death in cattle in East Anglia during the winters of 2001 to 2003. The condition had previously been limited mainly to stock imported into this part of Britain from endemically infected areas. In composite faecal samples collected by 16 farm animal veterinary practices in Norfolk, Suffolk and Essex, fluke eggs were found in 15 (28.8 per cent) of 52 previously unaffected suckler herds and 10 (16.7 per cent) of 60 dairy herds. Antibodies to F hepatica were detected by elisa in 32 (53.3 per cent) of the bulk milk samples from these 60 dairy herds, including the 10 in which fluke eggs were found. The emergence of fasciolosis in East Anglia was attributed to recent higher summer rainfall, which favoured the intermediate snail host Lymnaea truncatula and the free-living stages of F hepatica, the increased influx of sheep from endemic fluke areas for seasonal grazing, and the wetter grazing conditions associated with the Environmentally Sensitive Area scheme.     

An experimental study on triclabendazole resistance of Fasciola hepatica in sheep

The efficacy of triclabendazole in sheep experimentally infected with Fasciola hepatica was studied. Two groups of 12 lambs were infected with a susceptible (S) or a resistant (R) strain of F. hepatica. Eight weeks after infection, six lambs of each group (ST and RT) were treated with triclabendazole (10mg/kg). The other lambs were used as untreated controls (SC and RC). The parameters studied were: GLDH, gamma-GT, ELISA measuring antibodies against recombinant cathepsin-L(1) and eggs per gram faeces (epg). The lambs were slaughtered 16 weeks after infection and the number of flukes counted.The GLDH, gamma-GT levels and the OD value of the ELISA decreased as a result of the treatment in group ST. Patent infections were observed in all animals of groups SC, RT and RC. In group ST, occasionally a few eggs were found in five lambs. The percentage of flukes was 31.3 in SC and 37.6 in RC. In the treated groups ST and RT, the percentage of flukes was 0.06 and 33.6, respectively. These results corresponded to efficacies of 99.8% in the susceptible and 10.8% in the resistant strain. Since the resistant strain was isolated from a mixed cattle and sheep farm, it confirms the presence of triclabendazole resistance in the Netherlands.     

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.     

Fasciola in horses in the Republic of South Africa: a single natural case of Fasciola hepatica and the failure to infest ten horses either with F. hepatica or Fasciola gigantica

The faeces of 11 horses were examined for Fasciola spp. eggs. One of them was positive for Fasciola hepatica, a finding which was confirmed post-mortem. The 10 negative horses were subsequently infested with either F. hepatica or Fasciola gigantica, each animal receiving orally from 500-9,500 metacercariae. No clinical signs were observed and no fluke eggs were detected in the faeces, and neither immature nor adult Fasciola worms were recovered from the horses slaughtered 16-26 weeks post-infestation with F. hepatica and 28-34 weeks post-infestation with F. gigantica, respectively. The results indicate that horses have a high level of resistance to both South African Fasciola spp. Factors which can probably explain the conflicting reports for the prevalences of fascioliasis in horses throughout the world and the varying results obtained in experimental infestations are briefly considered.     

Relationships between host species and morphometric patterns in Fasciola hepatica adults and eggs from the northern Bolivian Altiplano hyperendemic region.

The highest prevalences and intensities of human fasciolosis by Fasciola hepatica are found in the northern Bolivian Altiplano, where sheep and cattle are the main reservoir host species and pigs and donkeys the secondary ones. Morphometric comparisons of many linear measurements, areas and ratios of F. hepatica adults (from sheep, cattle and pigs) and eggs (from sheep, cattle, pigs and donkeys) in natural liver fluke populations of the Bolivian Altiplano, as well as of F. hepatica adults and eggs experimentally obtained in Wistar rats infected with Altiplanic sheep, cattle and pig isolates, were made using computer image analysis and an allometric model. Although morphometric values of adult flukes from natural populations of sheep, cattle, and pigs showed great overlap, there were clear differences in allometric growth. The allometries analyzed were: body area (BA) versus body length (BL), BA versus body width (BW), BA versus perimeter (Pe), BA versus distance between posterior end of body and ventral sucker (P-VS), BL versus BW, BL versus Pe, and BL versus P-VS. These allometries show a good fit in the seven pairs of variables in all the populations examined. Comparative statistical analysis of the allometries shows that fluke adult populations from sheep, cattle and pigs significantly differ in BL versus BW and BL versus P-VS functions. Statistical analysis of F. hepatica egg size shows characteristic morphometric traits in each definitive host species. In experimentally infected rats, fluke adult allometry and egg morphometry do not vary depending on the Altiplanic definitive host species isolate. Our study reveals that the definitive host species decisively influences the size of F. hepatica adults and eggs, and these influences do not persist in a rodent definitive host model.     

Wasting and mortality in beef cattle parasitized by Eurytrema coelomaticum in the State of Paraná, southern Brazil

A sporadic wasting syndrome affecting beef cattle herds parasitized by Eurytrema coelomaticum is described in the State of Paraná, southern Brazil. The disease was characterized by progressive weight loss, poor body condition despite plenty of good quality forage available, and death. Annual losses ranged from 1 to 3%. The clinical course of the disease varied from 2 to 10 months after the onset of the first clinical signs. At necropsy, one of the three distinct following patterns of lesions were observed: the pancreas was of normal size and color; small, shrunken, white and markedly and diffusely firm (fibrosis); or slightly enlarged and dark with a shriveled capsular surface. Myriads of leaf-shaped trematodes of the genus Eurytrema were packed inside multiple dilated ducts with thickened, whitish fibrous walls, or the flukes were embedded in the remaining pancreatic parenchyma. Microscopic findings included extensive loss of the pancreatic parenchyma with replacement fibrosis, intralesional flukes and eggs, and ductal hyperplasia. Inflammatory reaction varied from absent to severe with the presence of a granulomatous reaction around the trematode eggs. One affected animal had high plasma amylase concentration (1580U/L) suggesting exocrine pancreatic insufficiency. Glucose blood levels were not significantly increased. We suggest that diffuse and marked fibrosis of the pancreas induced by the presence of large amounts of pancreatic flukes is the cause of the sporadic cases of chronic wasting and death of cattle in this geographic area.     

The anthelmintic efficacy of netobimin against experimental infections of Fasciola hepatica in sheep

Netobimin (coded SCH 32481, Schering Corporation), a new broad-spectrum anthelmintic having both fasciolicidal and nematocidal properties was evaluated for efficacy against mature Fasciola hepatica infections in sheep. The trial was conducted with 30 cross-bred spring lambs, each experimentally infected with 250 F. hepatica metacercariae. A single treatment of netobimin was administered at 17 weeks post-infection (PI) by oral drench at 7.5 or 20 mg kg-1 body weight while 10 animals remained as untreated controls. At necropsy (either 1 or 2 weeks post-treatment), the mean number of adult flukes recovered from the control, 7.5 and 20 mg kg-1 groups were 94.7, 35.9 and 8.8, respectively. The resulting efficacies were 62% (P less than or equal to 0.05) and 90.7% (P less than or equal to 0.01), respectively. No clinical signs of fascioliasis were noted in any sheep during the trial. No signs of toxicosis nor any adverse reactions to the drug were observed.     

Observations on helminth infections of free-living and captive rheas (Rhea americana) in Brazil

The present work describes helminth infection of eight free-living and 12 captive rheas (Rhea americana) from, respectively, Pantanal of Mato Grosso do Sul State, and Jaboticabal, Sao Paulo State, Brazil. Captive birds were young and had a high mortality rate, while free-living birds were adult and apparently healthy. Infections were evaluated by post-mortem examination of internal organs and recovery of helminths using standard parasitological procedures. Seven species of nematodes (Sicarius uncinipenis, Torquatoides crotophaga, Deletrocephalus dimidiatus, D. cesarpintoi, Paradeletrocephalus minor, Capillaria venteli and Dicheilonema rheae) and two species of cestodes (Houttuynia struthionis and Chapmania tauricolis) were identified. P. minor, which inhabits the large intestine, was the most common helminth in free-living birds (63.9%). In captive rheas, a mean parasitic load of 173 helminths per host was found. The gizzard of these birds was the most parasitized organ and S. uncinipenis was most common (92.5%). Parasitism of free-living and captive birds and associated pathology are discussed.     

Response of goats to repeated infections with Fasciola gigantica

One or two mature primary infections with Fasciola gigantica which had been removed by anthelmintic treatment resulted in a significant reduction in the number of flukes recovered from challenge infection as compared with that from controls.

Characteristic lesions of fascioliasis were seen in the livers of the 3 groups, however, goats with two primary abbreviated infections prior to challenge showed more severe lesions than those of animals with one primary abbreviated infection or those of challenge controls. The former group also showed the highest serum glutamate dehydrogenase and sorbitol dehydrogenase peaks following challenge infections and pulmonary fascioliasis was encountered in one of the goats of this group. Haemoglobin concentration and packed-cell volume decreased after infection in the three groups of goats.     

The effectiveness of the drugs Dovenix and Bilevon(R)-Injection against liver fascioliasis (Fasciola hepatica, Linné 1758) in cattle]

Dovenix and Bilevon-injection (manufactured by SPECIA, France and BAYER, West Germany, respectively) were tested for their anthelmintic efficacy against Fasciola hepatica in cattle. The drugs proved highly effective against both adult and immature flukes. The faeces of the treated animals were negative for F. hepatica eggs when examined 91 days after the treatment. In experiments with rats Dovenix and Bilevon-injection were tolerated up to eight times and ten times higher doses than the normal therapeutic dose, respectively.     

AN EPIDEMIOLOGICAL INVESTIGATION OF OVINE FASCIOLIASIS (FASCIOLA HEPATICA) ON BOTH IRRIGATED AND NON-IRRIGATED PASTURES IN NORTHERN VICTORIA

Tracer sheep were used to determine the temporal distribution of acquisition of F. hepatica on both irrigated and non-irrigated areas an area of northern Victoria where this parasite is endemic, for the period November 1974 to November 1976.
Throughout the study, very few flukes were acquired on the non-irrigated experimental area, and of this small number most were acquired by sheep in late spring and early summer. Flukes were acquired throughout both experimental years on the irrigated experimental area. The intensity of infection was lowest in late winter and early spring, but acquisition was substantial when measured over a year, being 483.5 in the first year and 1234.6 flukes per sheep in the second year.
The productivity of these animals was lower than that of equivalent animals kept on an area free of fluke, but quantitative interpretation of this information is confounded by the effect of the presence of an area with atypically high moisture levels (the snail habitat) on pasture production in the area grazed by the infected animals. The differences recorded are therefore likely to be underestimates of the true difference.
A suitable treatment regimen for control of liver fluke in sheep in northern Victoria is outlined.