Pulmonary embolization of immature Fascioloides magna causing fatal hemothorax confirmed by molecular technique in a heifer in the United States.

The current report describes the use of a molecular technique to identify immature Fascioloides magna An 18-month-old Brangus heifer was found dead in the field without any prior clinical signs. The cause of death was exsanguination into the thoracic cavity associated with pulmonary embolization and infection by immature Fascioloides magna resulting in 2 large foci of pulmonary necrosis and focal arteriolar and lung rupture. The liver had a few random migratory tracts with typical iron and porphyrin fluke exhaust, but no identified fluke larvae. A single immature fluke was found in the lungs, and species level identification as F. magna was confirmed by DNA sequence analysis of the ribosomal internal transcribed spacer regions (ITS1 region, 5.8S rRNA gene, and ITS2) and of partial 28S rRNA gene sequence. This is one of only a few pulmonary fascioloidiasis cases associated with hemothorax in the veterinary literature.

Multi level ecological fitting: indirect life cycles are not a barrier to host switching and invasion.

Many invasive species are able to escape from coevolved enemies and thus enjoy a competitive advantage over native species. However, during the invasion phase, non-native species must overcome many ecological and/or physiological hurdles before they become established and spread in their new habitats. This may explain why most introduced species either fail to establish or remain as rare interstitials in their new ranges. Studies focusing on invasive species have been based on plants or animals where establishment requires the possession of preadapted traits from their native ranges that enables them to establish and spread in their new habitats. The possession of preadapted traits that facilitate the exploitation of novel resources or to colonize novel habitats is known as 'ecological fitting'. Some species have evolved traits and life histories that reflect highly intimate associations with very specific types of habitats or niches. For these species, their phenological windows are narrow, and thus the ability to colonize non-native habitats requires that a number of conditions need to be met in accordance with their more specialized life histories. Some of the strongest examples of more complex ecological fitting involve invasive parasites that require different animal hosts to complete their life cycles. For instance, the giant liver fluke, Fascioloides magna, is a major parasite of several species of ungulates in North America. The species exhibits a life cycle whereby newly hatched larvae must find suitable intermediate hosts (freshwater snails) and mature larvae, definitive hosts (ungulates). Intermediate and definitive host ranges of F. magna in its native range are low in number, yet this parasite has been successfully introduced into Europe where it has become a parasite of native European snails and deer. We discuss how the ability of these parasites to overcome multiple ecophysiological barriers represents an excellent example of 'multiple-level ecological fitting'.

A comparative study of karyotypes and chromosomal location of rDNA genes in important liver flukes Fasciola hepatica and Fascioloides magna (Trematoda: Fasciolidae).

Chromosomal characteristics, i.e., number, size, morphology, and location of ribosomal DNA (rDNA) clusters were examined in two medically important liver flukes, Fasciola hepatica and Fascioloides magna (Fasciolidae), using conventional Giemsa staining and fluorescent in situ hybridization (FISH) with ribosomal 18S rDNA probe. A comparison of F. magna and F. hepatica karyotypes confirmed significant differences in all chromosomal features. Whilst the karyotype of F. hepatica comprised ten pairs of chromosomes (one metacentric and nine medium-sized subtelocentrics and submetacentrics; 2n = 20, n = 1 m + 5 sm + 4 st; TCL = 49.9 µm), the complement of F. magna was composed of 11 pairs of medium-sized subtelocentrics and submeta-metacentrics (2n = 22, n = 9 st + 1 sm + 1 sm-m; TCL = 35.2 µm). Noticeable differences were found mainly in length and morphology of first chromosome pair. It was metacentric and 9.0 µm long in F. hepatica while subtelocentric and 4.7 µm long in F. magna. Although FISH with rDNA probe revealed a single cluster of ribosomal genes in both species, conspicuous interspecific differences were displayed by chromosomal location of ribosomal loci (i.e., NORs). The signals were found on short arms of fifth homologous pair in F. hepatica; however, they were detected in pericentromeric regions of the long arms of tenth pair in F. magna. The observed cytogenetic differences were interpreted in terms of karyotype evolution of fasciolid flukes; F. hepatica may be regarded phylogenetically younger than F. magna. The present paper provides a pilot study on molecular cytogenetics within a group of hermaphroditic digenetic flukes.

Natural infection of a horse with Fascioloides magna.

A 25-year-old Quarterhorse mare was euthanized for a variety of medical reasons. At necropsy, 7 liver flukes, identified as Fascioloides magna, were recovered from the liver. This is the first report of F. magna in a horse.

[Fascioliasis case in the patient with hepatitis A].

There is some portion of patients with clinically manifested acute viral hepatitis, which are seronegative to hepatitis A markers. They have to be differentiated with other patients with B, C, D hepatitis, mechanical jaundice, etc. Such clinical cases make physician to recall the parasitic diseases, such as fascioliasis, which affects hepatobiliary system, causes prolongation of cholestasis and dystrophic changes in the biliary tract and likely to cause liver cirrhosis. In the presented case the initial diagnosis was severe acute Hepatitis A (anti-HAV IgM+), though the peripheral blood examination showed moderate eosinophilia, ultrasound investigation revealed multiple sites of damage in the liver, which made us to consider fascioliasis, the latter was confirmed by the serological analysis. Appropriate medical treatment was effective and the state of the patient has improved.

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.

Experimental Fascioloides magna infections of mule deer (Odocoileus hemionus hemionus).

Six mule deer (Odocoileus hemionus hemionus) and one white-tailed deer (Odocoileus virginianus), approximately 5-mo-old, each were inoculated orally with 500 metacercariae of Fascioloides magna. All mule deer died from liver fluke infection between 69 and 134 days (mean = 114, SE = 9.9) after inoculation. Between 38 and 326 immature F. magna (mean = 102, SE = 45.5) were recovered from each deer at necropsy. Flukes were present in livers, lungs, and free in pleural and peritoneal spaces. Infection was characterized by necrotizing hepatitis, fibrosing peritonitis and pleuritis, and hematin pigment accumulation in liver, lung, and many other internal organs. Eggs of F. magna first were detected in feces of the white-tailed deer 28 wk after inoculation, and weekly thereafter until the healthy deer was euthanized at 31 wk. At necropsy, 205 F. magna, including 12 encapsulated mature and 193 nonencapsulated immature flukes were recovered from liver, lungs, and free in abdominal and thoracic spaces of the white-tailed deer. Based on these results, F. magna may be fatal to mule deer within 5 mo of infection. Like domestic sheep and goats, mule deer may be highly susceptible to infection, and it is unlikely mule deer can survive infection with large numbers of F. magna.

Hepatic and pulmonary pathology of experimental Fascioloides magna infection in guinea pigs.

The guinea pig was used to study the pathology of Fascioloides magna, an important pathogen for sheep. Although flukes migrated freely through various tissues in infected guinea pigs, the most serious lesions occurred in the liver and lungs. The sequential development of lesions indicated that flukes first invaded the quadrate lobe of the liver and subsequently migrated to other liver lobes and tissues. Six weeks post-infection, there was a marked drop in the recovery of flukes from the liver along with a dramatic increase in pulmonary involvement. Much of the hepatic and pulmonary pathology in infected animals was secondary to extensive vascular lesions caused by migrating flukes. In the liver, vascular lesions predominantly involved the portal and hepatic veins. Thrombophlebitis and locally extensive necrosis, resembling infarction, were observed. Vascular lesions in the lungs occurred in the pulmonary arteries leading to thrombosis and haemorrhagic infarction. Discovery of a fluke in a pulmonary artery, along with the pattern of hepatic and pulmonary lesions, suggested that flukes probably used the cardiovascular system as a pathway for dissemination. Death in fluke-infested guinea pigs was most often associated with severe pulmonary lesions. The nature and distribution of fluke-induced lesions observed in this study demonstrate that the guinea pig is a suitable animal model for Fascioloides magna infection in sheep.

Domestic sheep as a rare definitive host of the large American liver fluke Fascioloides magna.

Two domestic sheep were inoculated orally with 250 metacercariae of Fascioloides magna. One sheep died 4 mo after inoculation and 22 immature F. magna were recovered. The second sheep remained healthy and was killed 8 mo after inoculation. Ten mature F. magna were recovered from hepatic parenchyma, and eggs of F. magna were observed in areas of the liver where flukes resided and from feces. Although F. magna is almost always lethal to sheep, this sheep was a rare definitive host.

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.

A natural infection of Fascioloides magna in a llama (Lama glama).

A young, female llama (Lama glama) was euthanized following the onset of hindleg paresis and paralysis. Live trematodes, identified as Fascioloides magna, were identified from the liver of this animal. This represents the first report of F. magna in a llama.

Efficacy of albendazole against Fascioloides magna in sheep.

Sheep experimentally infected with Fascioloides magna were used in a controlled study to determine the flukicide activity of different doses of albendazole (methyl [5(propylthio)-1H-benzimidazol-2-yl] carbamate). Sheep, about 4 months old, were inoculated with 100 metacercariae of F magna and treated with the various doses of albendazole 10 weeks later. At 6 weeks after treatment, the sheep were necropsied, flukes were recovered and measured, and the pathologic changes were recorded. The sheep were given different doses of albendazole (5, 7.5 [single dose or double dose], 10, and 15 mg/kg) and were compared with both nontreated inoculated and nontreated noninoculated controls. There were up to 70% fewer worms recovered in the group given 2 doses of 7.5 mg/kg than there were in the nontreated controls. There was also a significant reduction in flukes recovered in the groups given the 7.5, 10, and 15 mg/kg doses. Because a single F magna fluke has the potential to kill a sheep, it is imperative to consider the number of sheep with no flukes after treatment. According to that criterion, 50% of the sheep were protected at dose rates of 7.5 (single dose, double dose), 10, and 15 mg/kg of body weight.

Pathogenicity of immature Fascioloides magna in white-tailed deer.

The pathogenesis of early prepatent Fascioloides magna infection was investigated in seven fawns (Odocoileus virginianus) given 500 metacercariae and examined at one, two, three, five, eight, 12 and 13 weeks postinoculation. Blood samples were taken from eight inoculated deer every two weeks up to 16 weeks postinoculation. Eosinophilia with a mild transitory anemia were the main clincopathological features. Postmortem examination at two weeks postinoculation revealed extensive migration of immature flukes. Subcapsular tracks in the liver, nodules on the blind sacs of the rumen, as well as retroperitoneal granulomas on flanks and necrotic tracks on the diaphragm were found. Evidence of penetration of flukes into the lung was found at two weeks postinoculation and there was early granuloma formation at three weeks postinoculation. Flukes migrating into tissues other than the liver were destroyed in large granulomas, although remnants of degenerating parasites were not found. At eight weeks postinoculation, widespread granuloma formation characterized the infection with this lesion present in nodes along the gastrointestinal tract, in the mesentery, flanks, psoas muscles, diaphragm, between the ribs and in the lungs. By 12 weeks postinoculation subcapsular tracks were observed in the liver.

Experimental infection of domestic goats with Fascioloides magna.

Development of the deer liver fluke, Fascioloides magna, was studied in seven domestic goats, each experimentally inoculated with 250 metacercariae. Six infected goats died between 89 and 195 days (mean 139) after inoculation. Infection did not develop in the seventh goat. Mortality was the result of unrestricted fluke migration before maturation or encapsulation of the flukes. A total of 144 F magna (mean intensity of 24.0) were recovered from inoculated goats, representing 8.2% of the metacercariae administered. Growth rate of F magna in goats was similar to that in cattle, white-tailed deer, and sheep.

Selected clinicopathologic changes associated with experimentally induced Fascioloides magna infection in white-tailed deer.

Six white-tailed deer (Odocoileus virginianus), less than one year of age, were divided into two groups of three each and administered 50, or 500 metacercariae of ascioloides magna. All six deer became infected. Three additional deer of the same age were uninoculated controls. All deer were monitored for up to 43 weeks after inoculation to investigate changes in weight, selected hematologic values, and blood chemistry values. Although clinical disease was not evident in the infected deer, a significant reduction (p less than .01) in hemoglobin and packed cell volume was detected throughout the experiment. A significant elevation (p less than .01) in the total serum protein level was detected in both infected groups from 0 to 5 months after inoculation. Increases were present in the beta and gamma globulin fractions. No differences (p greater than .05) were detected in the serum calcium, magnesium, or phosphorus levels, or in body weights between infected and uninfected control groups.

Fascioloides magna in white-tailed deer (Odocoileus virginianus): observations on the pairing tendency.

Fascioloides magna (Bassi 1875) was recovered from 227 of 312 (73%) white-tailed deer (Odocoileus virginianus) in southern Texas between 1971 and 1975. Deer from age classes more than 1 year old had infection rates of from 64 to 84%. Few dead flukes were found. Flukes occurred exclusively in the livers of infected animals and were paired in 256 of 301 (85%) fibrous hepatic capsules. The prevalence of immature flukes with an average of one immature per infected liver was similar in all age classes, suggesting a relationship between fluke pairing and maturation.