Adult Fasciola Gigantica Research Articles

Some parasitic and bacterial causes of liver affections in ruminants

In the present study, a total number of 1576 livers of ruminants, including cattle (924), sheep (487) and camels (165) were examined for detection of different pathogenic agents. Among those, a number of 58 cases (3.68 %) were infected with both parasites and bacteria. Out of this number, 3 cases only (0.19 %) were infected with bacterial species (Lactobacillus lactis lactis), 31 cases (1.97 %) were infected with parasitic stages only and a number of 24 cases (1.52 %) were co-infected with both bacteria and parasites. Among the bacterial isolates, 11 genera of bacterial species were found in 27 cases. The obtained results revealed that, all infected cases were cattle. The most prevalent isolate was Enterococcus species which present in 7 cases (25.92 %) among the totally bacterial infected cases, followed by 5 cases showed Staphylococci (18.52 %), then both genus Escherichia and genus Lactobacillus, each was found in 4 cases (14.81 %) and finally 7 genera including;Enterobacter,Micrococcus,Citrobacter,Aerococcus,Pseudomonas,Chrysomonas and Streptococcus; each was found in a single case with a percentage of 3.70 %. Concerning the parasitic affections, it has been found that Fasciola species was the most prevalent type of helminthes present in the infected livers. Among all infected livers, 40 cases (2.54 %) were infected with adult Fasciola gigantica (including 33 cattle and 7 sheep), followed by 8 cases of camels (0.51 %) infected with Hydatid cysts, then 4 cases of sheep (0.25 %) infected with Cysticercus tenuicollis and finally 3 cases of sheep (0.19 %) infected with both Fasciola gigantica and Cysticercus tenuicollis. The results revealed that, a considerable high number of co-infected animals {24 cases (41.38 %) among the totally infected animals} was existed and this reveals that the parasite facilitates the way to the secondary bacterial infection with the end results of hepatic tissue degeneration and necrosis. So, in abattoirs, great attention must be taken for appropriate evaluation of these pathogenic sources.

Fasciola gigantica: Evaluation of the effect of phenyl vinyl sulfone in vitro

Phenyl vinyl sulfone is a synthetic inhibitor of cysteine protease and has antihelminthic and antiprotozoal properties. Phenyl vinyl sulfone was assayed in vitro for antifasciola activity against adult Fasciola gigantica worms using a well-established culture medium. Worms were treated with phenyl vinyl sulfone for incubation periods ranging from 0 to 12h and its activity was assessed in terms of viability, motility and death of worms. Phenyl vinyl sulfone exhibited a minimum effective concentration of 50ppm after 12h. Three hundred parts per million concentrations were most potent causing immediate death of adult flukes in vitro. Histopathological studies showed that there was tegumental flattening, rupture of vesicles, and spine loss. Marked reduction in size and number of ova and sperms in the convoluted tubules of the reproductive organs was observed in comparison to the untreated control group. In conclusion, phenyl vinyl sulfone shows potent activity against F. gigantica in vitro, and the authors recommend carrying out more studies to detect its efficacy in vivo.

Distribution of 28.5 kDa antigen in the tegument of adult Fasciola gigantica

Monoclonal antibody (MoAb) specific to 28.5 kDa tegumental antigen (TA) was used to localize this antigen in various tissues of adult Fasciola gigantica by means of indirect immunofluorescence, immunoperoxidase and immunogold techniques. The indirect immunofluorescence and immunoperoxidase detections revealed that this antigen was concentrated in the tegument particularly in its outer rim, tegumental cells and their processes, epithelial linings of the oral sucker and the proximal part of digestive tract. It was also detected at a moderate concentration in spermatogenic cells in the testes, cells of Mehlis’ gland, oocytes within the ovary, and ovum within the egg of adult parasites. At TEM level, the immunogold detection showed deposit of gold particles specifically in G 2 tegumental granules and on the surface membrane. Thus, this antigen is expressed in the tegument and associated structures of adult parasites, and it could be a major component of the G 2 granules which are shown to fuse with the surface membrane and contribute material to replace the casted-off membrane. This process is a part of membrane turnover that prevents the parasite from being attacked by the host immune effector cells.

Classification of the parenchymal cells in Fasciola gigantica based on ultrastructure and their expression of fatty acid binding proteins (FABPs)

Parenchymal cells in adult Fasciola gigantica can be classified into three types based on their ultrastructural features and different quantities of fatty acid binding protein (FABP) being stored. Parenchymal cell type 1 (Pc 1) has pale cytoplasm consisting largely of a loose network of fine fibers, and it contains few mitochondria but numerous glycogen particles. This cell type may be specialized in the storage and metabolism of glycogen and glucose. Parenchymal cell type 2 (Pc 2) has similar cytoplasmic features as Pc 1 but contains more numerous mitochondria, and high concentration of FABP as reflected by high density of immunostaining and immunogold labeling using specific monoclonal antibody (MoAb) to FABP as probe. Pc 2 may, thus, specialize in the storage and metabolism of fatty acids and other lipids. Parenchymal cell type 3 (Pc 3) has dense cytoplasm containing large amount of rough endoplasmic reticulum, Golgi complex and mitochondria, which is typical of a secretory cell. Furthermore, Pc 3 has very little glycogen particles and is not stained by MoAb against FABP. It could, thus, be concerned with the synthesis of fibers, which form the scaffold of the parenchyma.

Immunolocalization of cytoskeletal components in the tegument of the 3-week-old juvenile and adult Fasciola gigantica

Components of three cytoskeletal elements, namely, microtubule, intermediate and actin filaments have been localised in the tegument of the 3-week-old juvenile and adult Fasciola gigantica by means of immunofluorescence and immunoperoxidase techniques, using mouse monoclonal anti-alpha-tubulin, anti-cytokeratin antibodies and biotinylated-phalloidin, respectively. The immunostainings with the above probes were also performed in adult Schistosoma mansoni for comparison. The presence of tubulin, indicative of microtubules, was demonstrated in the tegumental cell bodies, their cytoplasmic processes, and the basal layer of the tegumental syncytium of F. gigantica. While in S. mansoni, tubulin appeared as vertical lines stretching across the whole thickness of the syncytium. Cytokeratin, representing one type of intermediate filaments, was detected in the tegumental cell bodies, their cytoplasmic processes, tegumental syncytium and spines of F. gigantica. In contrast, cytokeratin was evident only in the syncytium of S. mansoni, but not in the spines. Phalloidin, which could bind to actin, a subunit of microfilament, was detected in the tegumental cell bodies, their processes, and the microtrabecular network which form the scaffold of the tegumental syncytium of F. gigantica. In S. mansoni, actin was localized in similar tissues except the syncytium was not stained while spines exhibited intense staining. In F. gigantica, the presence of microtubules and actin filaments in the tegumental cells, their processes and in the syncytium could mediate the movement of secretory granules from the cell bodies towards the basal as well as the apical layer of the tegument. Cytokeratin filaments may serve to reinforce the integrity of the tegumental syncytium as well as the spines.

SERODIAGNOSIS OF HUMAN FASCIOLIASIS BY A CYSTATIN CAPTURE ENZYME-LINKED IMMUNOSORBENT ASSAY WITH RECOMBINANT FASCIOLA GIGANTICA CATHEPSIN L ANTIGEN

Cathepsin L1, a cysteine protease secreted by the gastrodermis of juvenile and adult Fasciola gigantica, was expressed in Escherichia coli as a calmodulin binding peptide fusion protein with a molecular mass of approximately 35 kD. The recombinant cathepsin L1 (rCTL1) was tested for its antigenic potential in a cystatin capture enzyme-linked immunosorbent assay (ELISA) to diagnose human fascioliasis. The ELISA plates were sensitized with chicken egg cystatin and incubated with bacterial lysates containing the recombinant protein before the standard ELISA procedures were performed. Analysis of the sera of 13 patients infected with F. gigantica (group 1), 204 patients with other parasitic infections (group 2), 32 cholangiocarcinoma patients (group 3), and 42 healthy controls (group 4) showed that the sensitivity, specificity, accuracy, positive predictive value, and negative predictive value of this ELISA using rCTL1 were 100%, 98.92%, 98.97%, 81.25%, and 100%, respectively. These results indicate that this assay has high sensitivity and specificity in the diagnosis of human fascioliasis. In addition, we have produced sufficient amounts of antigen for use in diagnosis.

Characterisation of Fasciola gigantica adult 27-kDa excretory-secretory antigen in human fascioliasis

The 27-kDa excretory-secretory antigen partially purified from adult Fasciola gigantica adult worms (FG27) has been used as the sensitive and specific antigen for immunodiagnosis of human fascioliasis. Lectin-blot analyses have shown that the FG27 possesses both N - and O -glycans. In two-dimensional electrophoresis, silver staining, lectin blotting and immunoblotting, FG27 displayed at least four antigenic-glycosylated protein spots at the pI values of 4.8, 4.9, 5.2 and 5.4, respectively. After removing glycan moieties from the antigen by alkaline treatment, the molecular mass of the FG27 was reduced to 26.5 kDa as a prominent deglycosylated band. Immunoblotting analysis has revealed that the 26.5-kDa band reacts with the pooled human fascioliasis sera without any loss of antigenicity. These results suggest that the major antigenicity of the FG27 is due to its protein epitopes.

Fasciola gigantica: surface topography of the adult tegument.

Adult Fasciola gigantica are leaf-shaped with tapered anterior and posterior ends and measure about 35 mm in length and 15 mm in width across the mid section. Under the scanning electron microscope its surface appears rough due to the presence of numerous spines and surface foldings. Both oral and ventral suckers have thick rims covered with transverse folds and appear spineless. On the anterior part of the ventral surface of the body, the spines are small and closely-spaced. Each spine has a serrated edge with 16 to 20 sharp points, and measures about 20 microm in width and 30 microm in height. In the mid-region the spines increase in size (up to 54 microm in width and 58 microm in height) and number, especially towards the lateral aspect of the body. Towards the posterior end the spines progressively decrease in both size and number. The tegumental surface between the spines appears highly corrugated with transverse folds alternating with grooves. At higher magnifications the surface of each fold is further increased with a meshwork of small ridges separated by variable-sized pits or slits. There are three types of sensory papillae on the surface. Types 1 and 2 are bulbous, measuring 4-6 microm in diameter at the base with nipple-like tips, and the type 2 also have short cilia. Type 3 papillae are also bulbous and of similar size but with a smooth surface. These sensory papillae usually occur in clusters, each having between 2 and 15 units depending on the region of the body. Clusters of papillae on the lateral aspect (usually types 1 and 2) and around the suckers (type 3) tend to be more numerous and larger in size. The dorsal side of the body exhibits similar surface features, but the spines and papillae appear less numerous and are smaller. Corrugation and invaginations of the surface are also less extensive than on the ventral side of the body.

Circulating antibodies and antigens correlate with egg counts in human fascioliasis.

We explored the relationships between specific IgG antibody levels and circulating E/S antigen to intensity of Fasciola infection in the human host. Twenty patients with patent infection and six healthy individuals were enrolled in the study. Intensity of infection was determined by repeated egg counts in stools, while IgG antibodies against adult Fasciola gigantica somatic FI, FII and against E/S antigens were measured as ELISA O.D. readings. The level of circulating E/S antigens was determined by IELISA. Positivity as well as levels of antibodies and antigen correlated with infection intensity. These findings may disclose in the future a relation between morbidity in the acute phase and worm load.

Fasciola gigantica-specific antigens: purification by a continuous-elution method and its evaluation for the diagnosis of human fascioliasis.

Immunodominant antigens of an approximate molecular mass of 27 kD were obtained from an excretory-secretory product of adult Fasciola gigantica by a continuous-elution method. An indirect ELISA using the antigens obtained by this relatively simple procedure was developed for detecting specific antibodies from patients infected with F. gigantica. Sera from patients with other parasitic infections, healthy volunteers, and cholangiocarcinoma were also analyzed. The sensitivity, specificity, and positive and negative predictive values for this ELISA using the fractionated antigens were 100%. The data indicated a possible correlation of antibodies to F. gigantica with cholangiocarcinoma.

Characterisation of protein antigen from Fasciola gigantica of different age

The protein antigens extracted from adult fluke Fasciola gigantica , 3, 6 and 9 weeks old and newly excysted juvenile (NEJ) were identified using SDS-PAGE and immunoblotting techniques. Sera from fat-tailed sheep which artificially infected with the metacercariae of F. gigantica were used for immunoblotting. The results showed that the protein antigen profile of adult fluke, 6 and 9 weeks old flukes had similar. Bands with molecular weight between 24 kDa to 114 kDa. Protein bands with molecular weight 198 kDa were also detected from the adult fluke. The use of immunoblotting technique, there were two antigenic protein molecules identified from adult fluke, NEJ and 3, 6, and 9 weeks old fluke with the molecular weight 46 kDa and 47 kDa. The protein band with molecular weight >198 kDa shown thicker on the NEJ than that of adult fluke, and 6 and 9 weeks old flukes. The role of protein with molecular weight of 46 and 47 kDa were the interested findings need to be evaluatedfor serological analysis. Key words : Protein antigen, Fasciola gigantica , SDS-PAGE, immunoblotting

Evaluation of antigens of Fasciola gigantica as vaccines against tropical fasciolosis in cattle

Vaccine trials were conducted in Brahman cross cattle evaluating the efficacy of 4 native antigens purified from adult Fasciola gigantica flukes, and 1 recombinant F. gigantica antigen, as vaccines against tropical fasciolosis. The antigens tested were native glutathione S-transferase, cathepsin L, paramyosin, fatty acid binding protein (FABP), and a recombinant FABP expressed in E. coli, and were formulated in 1 or more of several adjuvants (Quil A, Squalene Montanide 80 ®, MF59-100, Auspharm ®, NAGO, polylactoglycolide microspheres, Algammulin, DEAE, Freund's). Vaccination induced low, moderate or high antibody titres to the various antigens which were dependent on the adjuvant. Low but significant reductions in fluke burdens (31%, P < 0.026) and fluke wet weight (36%, P < 0.041) were only observed in cattle vaccinated with the native FABP in Freund's adjuvant. There was no correlation between total antibody titres to FABP and protection. The protection observed in cattle vaccinated with native FABP of F. gigantica supports the notion that this class of proteins is a useful target for protection of animals against Fasciola and extends the efficacy of FABPs to the tropical liver fluke. This is the first report of vaccination of cattle against F. gigantica with a purified protein.

Partial purification and characterisation of the proteolytic enzymes of Fasciola gigantica adult worms

The proteases of adult Fasciola gigantica whole worm were analysed by preparative isoelectric focusing and by gelatin-substrate gel electrophoresis at acidic and neutral pH (4.5 and 7.0). At least 15 bands of proteases were observed. These proteases had molecular weights ranging from 26 to 193 kDa and isoelectric points of 4.92–7.63. Potease-rich fractions were subsequently separated from whole worm preparation of the parasite by filtration in Sephacryl S-200. The proteases were able to digest bovine immunoglobulin G (IgG) and globin (derived from bovine haemoglobin) in vitro. The sizes of the proteases in these fractions were from 26 to 96 kDa, and they were inhibited by the protease inhibitors phenylmethylsulphonyl fluoride (PMSF), leupeptin and trasylol.

Première observation de l’hépatite infectieuse nécrosante (black disease) parmi des chèvres nubiennes au Soudan

In a flock of 425 female and male Nubian goats in the Khartoum Province, an outbreak of a disease causing sudden death of 18 apparently healthy goats occurred (11 females and 7 males, 3-6 years old). Adult Fasciola gigantica were found in the livers of all goats and in seven of them Cysticercus tenuicollis cysts. These organs showed necrotic and severe histopathological changes. Clostridium novyi type B was isolated from necrotic areas of all livers and found to be highly pathogenic and toxigenic to laboratory animals. The disease was diagnosed as infectious necrotic hepatitis (black disease). Faecal examination revealed the presence of F. gigantica eggs. Lymnaea natalensis snails were found to be prevalent in the water canals. As the Khartoum Province is regarded as an endemic area for black disease, routine vaccination is highly recommended for its control in goats and sheep.

Serodiagnosis of fascioliasis in buffaloes by immunoprecipitation using adult Fasciola gigantica antigens

Partial purification of a saline extract of adult Fasciola gigantica by chromatography on Sephadex G200 gave four peaks, of which only the first two contained material which gave a precipitation reaction with sera from infected buffaloes. None of these gave false reactions with sera from uninfected animals but even the first and most sensitive fraction only reacted with 57.5 per cent of sera from animals known to be infected with moderate numbers of F gigantica.

Carbohydrate energy metabolism in Fasciola gigantica (trematoda)

Umezurike G. M. and Anya A. O. 1980. Carbohydrate energy metabolism in Fasciola gigantica (Trematoda). International Journal for Parasitology 10: 175–180. Adult Fasciola gigantica contained 4.49 ± 0.06 % (mean ± S.D.) wet weight glycogen. Tissue homogenates contained high levels of malate dehydrogenase (MDH), NAD-linked malic enzyme (ME), Phosphoenolpyruvate carboxykinase (PEPCK) and lactate dehydrogenase (LDH). MDH, PEPCK and ME activities appeared to be localized in both cytosolic and mitoehondrial fractions, fumarase activity appeared to be predominantly mitochondrial whereas LDH and pyruvate kinase activities were cytosolic in distribution. Polyacrylamide gel electrophoresis revealed the predominance of LDH-1, LDH-2 and LDH-3 but only traces of LDH-4 and LDH-5 isoenzymes in the crude cytosolic fraction. LDH activity in the crude sample was inhibited by excess substrate (pyruvate). The mitoehondrial system showed NADH -cytochrome c oxidoreductase, succinate-cytochrome c oxidoreductase, NADH oxidase and some cytochrome c-oxygen oxidoreductase activities. Under anaerobic conditions, NADH-fumarate oxidoreductase and succinate-NAD + oxidoreductase activities of mitoehondrial preparations were stimulated in the presence of ADP and ATP respectively. Isolated mitochondria contained rhodoquinone and no ubiquinone, and isolated rhodoquinone was readily reduced by succinate in the presence of submitochondrial particles. Hydrogen peroxide was produced by submitochondrial particles in the presence or absence of KCN or in the presence of fumarate.

Preliminary Observations on Antigen Activity of Fasciola Gigantica by Immunodiffusion Test in Sheep

Antigen prepared from adult Fasciola gigantica was precipitated in agar with sera obtained from lambs infected experimentally with F. gigantica. The agar gel precipitin test appeared to be a useful technique in the diagnosis of F. gigantica infection in sheep. Precipitins were detected as early as 3 to 4 weeks after infection and it was also possible to recognize the infection after the appearance of eggs in faeces.

Prevalence of Fasciolia gigantica in slaughtered cattle in Ado municipal abattoir, Ado Ekiti, Ekiti state, Nigeria.

This study was carried out in the month of February and March 2014 to determine the prevalence and the intensity of Fasciola gigantica infection in slaughtered cattle in Ado municipal abattoir. The abattoir was visited twice in a week; the liver of cattle slaughtered were examined for presence of adult Fasciola gigantica. A total of 475 cattle were examined during the period, out of which eighty-one (17.1%) were positive for presence of Fasciola gigantica with varying levels of intensity. The month of March had the highest infection rate of 22.2% than February which was 12.2%. Males had the highest infection rate in the two months than the females. Statistical analysis revealed that there was no significant difference in the infection in males and females (P&gt; 0.05). This result established the presence of Fasciola gigantica in Ado Ekiti with prevalence rate of 17.1%.Among positive samples (n=81), 27 % (n=22) were light (0-10 flukes), 58 % (n=47) moderate (11=50 flukes), and 14 % (n=12) heavy intensity of infection. Cattle slaughtered at this abattoir are majorly purchased from the northern part of the country. Grazing of these cattle on pasture visited by local flock’s leads to cross infection. Control measures should therefore be instituted, like routine anthelmintic administration and eradication of the snail intermediate host to curtail the spread of the parasite in the area. More surveillance should be mounted by Veterinary officers and public health officers against the disease in slaughtered cattle.Key words: Fasciola gigantica, Cattle, Liver, Abattoir, Ado Ekiti

Correlation between the results of intradermal and complement fixation tests in the diagnosis of urinary schistosomiasis

The results of the use of 10 antigens prepared from adult Fasciola gigantica, Schistosoma mansoni and Schistosoma bovis, used for intradermal and complement fixation tests, have been evaluated in the diagnosis of urinary schistosomiasis. A combination of the two tests with delipidized F. gigantica antigen is recommended.

PREVALENCE OF SCHISTOSOMIASIS IN VARIOUS GOVERNORATES IN THE UNITED ARAB REPUBLIC AS REVEALED BY THE INTRADERMAL TEST WITH ANTIGENS FROM ADULT FASCIOLA GIGANTICA AND SCHISTOSOMA MANSONI.

Abstract The incidence of schistosomiasis in eight different localities has been investigated. Subjects were examined by a urine sedimentation method, and by the intradermal test with antigens prepared from Fasciola gigantica and Schistosoma mansoni adult worms. The results show that this intradermal test is valuable in epidemiological surveys.