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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.

Animals↗

Triploid form of Fasciola in Japan: genetic relationships between Fasciola hepatica and Fasciola gigantica determined by ITS-2 sequence of nuclear rDNA.

The ITS-2 sequences from seven worms of the Japanese triploid form of Fasciola, two of Fasciola hepatica from Uruguay and four of Fasciola gigantica from Zambia and Indonesia were determined. The ITS-2 sequences of the Japanese triploid worms were divided into two distinct types, one of which was almost identical to that of F. hepatica and the other to F. gigantica from Indonesia.

Animals↗

The structure and cytochemistry of the neurosecretory cells of Fasciola gigantica Cobbold and Fasciola hepatica L.

Histochemical studies of the nervous system of Fasciola gigantica and Fasciola hepatica were undertaken. Neurosecretory cells were detected by Gomori's aldehydefuchsin, Bargmann's chrome hematoxylin-phloxin, Mallory's triple stain, periodic acid-Schiff, Heidenhain's Azan and alcian blue after potassium permanganate oxidation. Two types of neurosecretory cells were recognized and designated as "A" and "B". Type "A" cells occurred in small numbers in the brain and subesophageal mass and type "B" cells ubiquitous in distribution. The reactions of these cells to the standard stains for neurosecretory substance generally, were less intense than the neurosecretory cells of other animals such as crustaceans and insects. The structure, organisation, distribution and cytochemistry of neurosecretory cells in Fasciola gigantica and Fasciola hepatica is discussed.

Animals↗

Fasciola gigantica and F hepatica: a comparative study of some characteristics of Fasciola infection in Lymnaea truncatula infected by either of the two trematodes.

Experimental infections were carried out using three Lymnaea truncatula populations and two Fasciola species in order to determine the trematode influence on six parameters of snail infection by either of the two trematodes. All experiments were performed using snails 4 mm long, two miracidia for each L truncatula, and a constant temperature of 20 degrees C. No significant influence of trematode species was detected in the following parameters: snail survival at day 30, the life span of infected snails (from miracidial exposure to snail death), the increase in shell length throughout the experiment, the duration of the patent period and the number of cercariae shed by infected snails. The frequency of cercaria-shedding snails was closely correlated with the particular L truncatula population and trematode species. A similar finding in two populations was also noted for the frequencies found in the infected snails who died without emission. The prepatent period had a longer duration in the Fasciola gigantica-infected groups. The percentages of floating cysts were greater in the Fasciola gigantica groups than in the Fasciola hepatica groups.

Animals↗

Evaluation of circulating anti-Fasciola IgA antibodies by ELISA technique using two different Fasciola antigens.

Two types of Fasciola antigens: surface tegumental (ST) and excretory-secretory (ES) products of F. gigantica worms were used for performance of ELISA technique. Results showed that, the circulating anti-Fasciola IgA antibodies were detected in 98% and 94% and the specificity of the test was 93.4% and 89.7% using ST and ES antigens, respectively. As regards ST antigen, a highly significant elevation in the level of the circulating anti-Fasciola IgA antibodies was observed according to the intensity of infection as well as in those with hepatomegaly. There was a significant direct relation between the level of circulating anti-Fasciola IgA antibodies and eosinophilia, anaemia and liver function tests of fascioliasis patients when ST antigen was used.

Adolescent↗

Comparison of modulation of sheep, mouse and buffalo lymphocyte responses by Fasciola hepatica and Fasciola gigantica excretory-secretory products.

In order to explain the difference in susceptibility to Fasciola hepatica and F. gigantica between animal species, the activity of their excretory-secretory products (FhESP and FgESP, respectively) on concanavalin A (ConA)-induced proliferation of different animal species (sheep, mouse and buffalo) lymphocytes was compared. At high doses, FhESP inhibited proliferation of lymphocytes of all the animal species tested, and at low doses they inhibited the proliferation of sheep lymphocytes and increased the proliferation of buffalo and mouse lymphocytes. The effects of FgESP were similar but the intensity of FgESP inhibition was less than FhESP. The immunomodulatory effects of FhESP or FgESP could not alone explain the susceptibility level of hosts to Fasciola spp. The immunomodulatory molecules of FhESP and FgESP and their role in the course of Fasciola spp. infection should be further investigated.

Animals↗

Serum antibody isotype responses of Fasciola-infected sheep and cattle to excretory and secretory products of Fasciola species.

This study investigated the immunoglobulin isotype responses of sheep and cattle chronically infected with Fasciola hepatica and Fasciola gigantica to adult F. hepatica excretory/secretory products (Fh-ES) or F. gigantica excretory/secretory products (Fg-ES), respectively. An antibody enzyme-linked immunosorbent assay (Ab-ELISA) was used to determine serum antibody (total Ig, IgG(1), IgM, IgG(2) and IgA) responses. At necropsy, the mean number of flukes recovered was lower in cattle than in sheep. All F. hepatica and F. gigantica infected sheep and cattle showed an increased total Ig levels from 3 to 4 weeks post-infection (wpi). Among isotypes IgG(1) was most dominant while IgM was the earliest (2 wpi) to be detected in both sheep and cattle infected with both F. hepatica and F. gigantica animals. IgG(2) response was early (2 wpi) in sheep infected by F. hepatica but there was no response in sheep infected with F. gigantica. There was a late and strong IgG(2) response in cattle infected with both flukes. The IgA isotype showed an early and a clear biphasic response in sheep with F. hepatica but was less pronounced in F. gigantica infected sheep. While IgA response to Fh-ES was noticed 5 wpi in F. hepatica infected cattle, it appeared much later (21 wpi) in those infected with F. gigantica. The dominance of IgG(1) isotype in infected sheep and cattle suggest an associated Th2 response. This early response to adult Fasciola spp. ES antigen suggests an early exposure to the antigen presumably through the cross-reacting ES products of juvenile flukes. There is clearly difference in IgG(2) isotype response in cattle (resistant) compared to sheep (susceptible). The late IgG(2) response in cattle may suggest late Th1 involvement in bovine cellular responses to adult Fh-ES/Fg-ES.

Animals↗

Fasciola hepatica and Fasciola gigantica: comparative morphometric studies on the redial stage of both species.

Experimental infections of Galba truncatula with Fasciola gigantica or F. hepatica were carried out under laboratory conditions (20 degrees C) to determine the characteristics of rediae of both species via their morphometry and to find reliable measurements that might be efficiently used to discriminate between the rediae of both species of Fasciola. These results were compared to those of another snail: Radix natalensis, infected with either F. gigantica or F. hepatica under the same protocol. At day 28 post-exposure, abortive infections with F. hepatica were found in a group of R. natalensis. By contrast, live rediae were observed in the other three groups. The group of infected snails and the redial category significantly influenced the mean values of the seven measurements studied and those of three indices. Using the PSLD Fisher test, it was found that the index, distance from the anterior end of the body to the collar/length of the body, was an efficient means of distinguishing the rediae of F. hepatica from those of F. gigantica [second-appearing mother rediae (R1b) of the first generation, 0.14 instead of 0.22; daughter rediae (R2a) produced by the first mother rediae, 0.19 instead of 0.24]. Another index, distance from the anterior end of the body to the collar/diameter of the collar, could also be used to discriminate between rediae (R1b, 0.80 for F. hepatica instead of 1.09 for F. gigantica; R2a, 0.90 instead of 1.26, respectively). Compared to measurements recorded for the rediae of F. hepatica, rediae of F. gigantica can be characterized by the following measurements: the diameter of the pharyngeal lumen and the distance from the anterior end of the body to the collar for larvae developed in R. natalensis, and the length of the body and the distance from the posterior end of the body to lateral projections for those found in G. truncatula. The species of snail host and, consequently, its growth, as well as the species of Fasciola, had a significant influence on the morphometric characters of the redial stage.

Animals↗

Phenotypic comparison of allopatric populations of Fasciola hepatica and Fasciola gigantica from European and African bovines using a computer image analysis system (CIAS).

The quantification of the different sizes and shapes of Fasciola hepatica and Fasciola gigantica from bovines has been achieved for the first time in natural allopatric populations. Linear measurements, areas and ratios of gravid adults and eggs of F. hepatica (from France and Spain) and F. gigantica (from Burkina Faso) were analysed using a computer image analysis system and an allometric model: (y2m - y2)/y2 = c[(y1m - y1)/y1](b), where y1 = body area or body length, y2 = one of the measurements analysed, y1m, y2m = maximum values towards which y1 and y2, respectively, tend and c, b = constants. All the measurements overlap in the two fasciolids, apart from the distance between the ventral sucker and the posterior end of the body, body roundness and body length/body width ratio. The results obtained may be useful in Fasciola species identification in countries where both species coexist.

Africa↗

The effect of pre-exposure to Fasciola hepatica or Schistosoma mansoni on challenge infection with Fasciola hepatica.

Two groups of 12 and 6 rats were inoculated with Fasciola hepatica and Schistosoma mansoni, respectively. The Schistosoma-inoculated group, as well as 6 Fasciola-inoculated rats and 6 uninfected rats were challenged 8 weeks later with F. hepatica. A control group of 6 rats was left unexposed. Eight weeks after the challenge exposure all rats were necropsied and subjected to post-mortem examination. The number of Fasciola recovered after challenge was lower in both groups of rats primarily infected with F. hepatica or S. mansoni. F. hepatica-induced pathological changes were observed in all infected rats, but were pronounced in the group which was first exposed at the time of challenge of the primarily infected groups. No Schistosoma eggs or adults were detected in Schistosoma-inoculated rats. The results demonstrated that rats primarily infected with F. hepatica acquired resistance against a challenge exposure to the homologous parasite. Also S. mansoni, even without patency, can provide partial protection against F. hepatica infection.

Animals↗

A PCR-RFLP assay for the distinction between Fasciola hepatica and Fasciola gigantica.

Fascioliasis is of well-known veterinary importance and an increasing human health problem, with reported cases in the five continents. The causative agents, Fasciola hepatica and Fasciola gigantica, present geographical distributions, which overlap in many regions of Africa and Asia, and in which the differentiation of both species is usually difficult because of the many variations in their morphological characteristics. Moreover, in humans, liver fluke classification cannot be achieved by clinical, pathological, coprological or immunological methods. The differential diagnosis between F. hepatica and F. gigantica infection is very important because of their different transmission and epidemiological characteristics. A simple and rapid PCR-restriction fragment length polymorphism (RFLP) assay, using the common restriction enzymes Ava II and Dra II, is described to distinguish between both fasciolid species. It is based on a 618-bp-long sequence of the 28S rRNA gene recently obtained from liver fluke populations of South America, Europe and Africa. This sequence showed a few nucleotide differences between both fasciolids and no intraspecific variations within each species. This assay provides unambiguous results and may be useful for both individual subject diagnosis and epidemiological surveys of humans and animals in endemic regions of sympatry.

Animals↗

Fasciola hepatica and Fasciola gigantica: comparison of cellular response to experimental infection in sheep.

Cellular responses to Fasciola gigantica and to Fasciola hepatica infection in sheep were compared. Eosinophil numbers increased more quickly and strongly in F. gigantica-infected sheep than in F. hepatica-infected sheep. In both groups, peripheral blood mononuclear cell (PBMC) proliferation in response to the parasitic excretory-secretory products (ESP) showed similar kinetics. Interferon-gamma (IFN-gamma) production by ESP-stimulated PBMC was early and showed similar kinetics in both groups. Interleukin-10 (IL-10) production by FhESP-stimulated PBMC was very high throughout infection even at 0 weeks post-infection (WPI) in F. hepatica-infected sheep, while in F. gigantica-infected sheep, IL-10 production by FgESP-stimulated PBMC increased between 1 and 4 WPI. IL-10 production in F. gigantica-infected sheep was significantly lower than in F. hepatica-infected sheep during infection. The lower susceptibility to F. gigantica infection in sheep could be explained by the more intense cellular response induced by the parasite and the weaker capacity of F. gigantica to evade the immune response.

Animals↗

Phenotypic analysis of adults of Fasciola hepatica, Fasciola gigantica and intermediate forms from the endemic region of Gilan, Iran.

Fascioliasis is an important human and animal disease caused by Fasciola hepatica and Fasciola gigantica. In Iran, the distribution of these two species overlaps in most areas, including the northern human endemic province of Gilan where both fasciolids are simultaneously found in individual cattle and buffaloes. A phenotypic study of fasciolid adult flukes from naturally infected bovines from Gilan was carried out by means of an exhaustive morphometric analysis using traditional microscopic measurements and an allometric model. The Iranian fasciolids were compared to F. hepatica and F. gigantica standard populations, i.e. from geographical areas where both species do not co-exist (Bolivia and Burkina Faso, respectively). Although morphometric values somewhat overlapped, there were clear differences in allometric growth. The allometric function was adjusted to 25 pairs of variables. Results obtained revealed that Iranian F. hepatica-like specimens are larger than the F. hepatica standard and Iranian F. gigantica-like specimens are longer and narrower than the F. gigantica standard, but with smaller body area. Measurements which permit a specific differentiation in allopatric populations (distance between ventral sucker and posterior end of the body; ratio between body length and body width) overlap in the specimens from Gilan, thus proving the presence of intermediate forms. When compared to the standard populations, the different Iranian fasciolid morphs show greater differences in F. gigantica-like specimens than in F. hepatica-like specimens. This study shows that simple, traditional microscopic measurements may be sufficient for the morphometric characterisation of fasciolids, even in areas where intermediate forms are present.

Animals↗

The serum glucose and beta-hydroxybutyrate levels in sheep with experimental Fasciola hepatica and Fasciola gigantica infection.

The influence of Fasciola hepatica and Fasciola gigantica infection on serum glucose and beta-hydroxybutyrate (beta-HOB) in sheep was evaluated. This was done by setting up two groups of sheep. The first group (n=13) was split in two sub-groups, one experimentally infected with F. hepatica (n=9) and the other (n=4) as uninfected control. A second group consisting of a sub-group experimentally infected with F. gigantica (n=9) the other sub-group (n=6) left as uninfected control was also set up. The results of weight gain, parasitological and serum liver enzymes activity (glutamate dehydrogenase [GLDH] and gamma glutamyltransferase [gamma-GT]) used in monitoring the infection showed that all infected animals developed fasciolosis. It was observed that a reduction in serum glucose levels was significantly lower (p<0.05) in F. hepatica infected sheep than in uninfected control sheep starting from 5 weeks post-infection (wpi) to the end of the experiment. Similar reduction was recorded in F. gigantica infected sheep between 8 and 19 wpi. In contrast, serum beta-HOB levels were elevated in F. hepatica infected sheep between 6 and 16 wpi and in F. gigantica infected sheep between 7 and 15 wpi. It would appear from these serum glucose and beta-HOB levels that fasciolosis does lead to energy deficiency (low glucose) and ketosis (increased beta-HOB). The decrease in serum glucose and increase in serum beta-HOB levels in infected sheep may help in understanding the interaction between fasciolosis and nutritional status of infected ruminants especially in young growing animals.

3-Hydroxybutyric Acid↗

Host responses during experimental infection with Fasciola gigantica or Fasciola hepatica in Merino sheep I. Comparative immunological and plasma biochemical changes during early infection.

This study reports the early biochemical changes in plasma, comparative host-immune responses and parasite recovery data in Merino sheep during the first 10 weeks of infection with Fasciola gigantica and Fasciola hepatica. One group of sheep were uninfected, four groups of sheep received incremental challenge doses of F. gigantica metacercariae (50, 125, 225 and 400, respectively) and the sixth group was challenged with 250 F. hepatica metacercariae. At 10 weeks post infection (wpi), sheep challenged with F. hepatica showed the greatest fluke recovery (mean 119, range 84-166); a significantly higher biomass of parasites recovered (2.5-fold greater than the highest dose of F. gigantica); and a greater mean % parasite recovery (39.3%, range 27-55%) than any group challenged with F. gigantica. Within the groups dosed with F. gigantica a strong dose-dependent response was observed in both fluke recovery and fluke biomass with increasing dose of metacercariae. The mean % parasite recovery of F. gigantica infected groups 1-5 were 26, 23, 26 and 25%, respectively, suggesting a uniform viability of parasite establishment independent of infection dose. At 6 wpi, elevated levels of plasma GLDH were observed in the F. gigantica infected groups compared to the uninfected sheep (p<0.005) whereas the F. hepatica challenged group had four-fold higher levels of GLDH compared to the F. gigantica infected group (p<0.001). Elevated levels of GGT as an indicator of epithelial damage in the bile duct was only seen in the group challenged with F. hepatica at 10 wpi when it rose from below 100 IU/l to approximately 250 IU/l (p<0.0001) whereas no detectable increase in GGT was observed in any of the groups challenged with F. gigantica. The white blood cell response to F. hepatica infection was biphasic with the initial peak at 4 wpi and a second peak at 9 wpi, corresponding to the period of migration of juvenile fluke in the liver and the time when adult flukes are migrating into the bile duct, respectively. This biphasic response was also evident in the changes in the eosinophil counts and serum haemoglobin levels. There was a trend toward higher parasite-specific IgG2 titres in sheep infected with lower worm burdens, suggesting that higher F. gigantica or F. hepatica burdens suppress IgG2 responses. The findings of this study suggest that, in early infection in a permissive host, F. hepatica appears to be more pathogenic than F. gigantica because of its rapid increase in size and the speed of its progression through the migratory phases of its life cycle.

Animals↗

The liver flukes Fasciola gigantica and Fasciola hepatica express the leucocyte cluster of differentiation marker CD77 (globotriaosylceramide) in their tegument.

Glycosphingolipids from the parasitic liver flukes Fasciola gigantica and Fasciola hepatica were isolated and their carbohydrate moieties were structurally analysed by methylation analysis, exoglycosidase treatment, on-target exoglycosidase cleavage and matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry. For both liver fluke species, the ceramide monohexosides Gal1-ceramide and Glc1-ceramide were found in relative amounts of 1.0 to 0.1, respectively. From F. gigantica, the ceramide dihexoside was isolated in sufficient amounts to be structurally determined as lactosylceramide, Gal beta4-Glc1-ceramide, while for both liver fluke species the ceramide trihexoside was shown to be Gal alpha4Gal beta4-Glc1-ceramide, which is designated as either globotriaosylceramide, Pk-blood group antigen or CD77 leucocyte cluster of differentiation antigen. To our knowledge, this is the first report on the expression of globo-series glycosphingolipids in non-mammalian species. Ceramide analysis of ceramide monohexosides yielded as major components octadecanoic and 2-hydroxyoctadecanoic fatty acids together with C18- and C20-phytosphingosines. By the use of an anti-CD77 monoclonal antibody and the Escherichia coli Shiga toxin B1 subunit, globotriaosylceramide could be immunolocalised to the tegument of F. hepatica cryosections. The sharing of CD77 between liver flukes and their mammalian hosts fits in with the concept of molecular mimicry, which is closely parallel to the established imitation of host CD15 (Lewis X) displayed by the blood fluke Schistosoma mansoni.

Animals↗

Banding patterns of Fasciola hepatica and Fasciola gigantica (Trematoda) by isoelectric focusing.

Liver flukes Fasciola hepatica and Fasciola gigantica are polymorphic and vary morphologically depending upon the host being parasitized. It is known also that mixed infection occurs where both species are present. A technique involving protein separation was used to distinguish the 2 species. Isoelectric focusing of soluble proteins was performed on polyacrylamide gels using whole-body proteins from adult flukes. Although many bands appeared common to both species and some were shared with host tissues, the banding patterns could be used to distinguish 1 species from the other. Soluble protein isoelectric focusing is simple, reproducible, and has very good resolution. It seems well suited to the differentiation of the 2 fluke species.

Animals↗

Detection of genetic variabiltiy in nonhuman isolates of Fasciola hepatica and Fasciola gigantica by the RAPD-PCR technique.

The present study shows the molecular characterization of Fasciola gigantica and F. hepatica isolates collected from cows and sheep, using the random amplified polymorphic DNA fragments-polymerase chain reaction (RAPDs-PCR) technique. Optimal standardization of amplification conditions and thermocyclation were made, using genetic markers. The methodology used compared the genetic pattern between the two species (inter-species) and inside each species (intra-species) between cow and sheep and the amplification fragments were between 135 and 741 base pairs of marker. The results showed genetic variations (polymorphisms) of Fasciola gigantica and F. hepa-tica with amplification fragment based on a 500-400 base pair (bp). Inside each species, there were genetic variations in bovine and ovine and the amplification fragments were between 600 and 400 base pairs (bp). This assay is useful for both individual diagnosis and epidemiological surveys in endemic regions.

Animals↗