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Fascioliasis is an important zoonotic disease with approximately 2-4 million people infected worldwide and a further 180 million at risk of infection. F. hepatica can survive within the bile ducts for many years through its ability to suppress the host immunity with Fasciola cathepsin L1 cysteine protease and Glutathione S transferase playing an important role. The aim of the present study is to investigate the in vitro lympho-proliferative responses of hepatic hilar lymphocytes (HLN) of infected sheep in response to different F. hepatica antigens. The suppressive effects of Fasciola excretory/secretory (ES) and tegument (TEG) and their fractions were also investigated. Our results showed that both ES and TEG had significant suppressive effects on lympho-proliferation, up to 74% and 92%, respectively. When these antigens were fractionated, fraction 3 (MW of >10000-30000) of both ES (64%) and TEG (59%) in addition to fraction 4 (MW of ≤ 10000) of TEG (38%) inherited the suppressive effects. Identification of the potential molecule(s) with such suppressive effects on lymphocytes in TEG fraction 4 could reveal vaccine candidates.

Fasciolosis caused by Fasciola gigantica is one of the major public health problems in the world including Egypt. Immunodiagnostic methods are more applicable for their better sensitivity and specificity than other methods. The present study was conducted to cysteine proteinase (CP) antigens of F. gigantica in IgG-ELISA to diagnose human fasciolosis. IgG-ELISA with 2 cysteine proteinases of 27 kDa (Fas1) and 29 kDa (Fas2), obtained from the regurgitated materials of adult worms, were evaluated using serum samples from 90 Egyptian patients infected with F. gigantica, 55 patients with other parasitic infections and 50 healthy volunteers. The diagnostic sensitivity and specificity of Fas1 for detection of F. gigantica infection were 91.1% and 89.1%, respectively. The positivity of the assay was 95%. The positive and negative predicted values were 91% and 86%, respectively. These data suggest that IgG-ELISA with Fas1 is highly sensitive and specific assay and could be used for the immunodiagnosis of human fasciolosis.

Identification of liver fluke species cannot be achieved by clinical, pathological, coprological or immunological methods. However, the differential diagnosis between F. hepatica and F. gigantica infection is very important because of their different pathological manifestations. Moreover, in countries where the two species co-exist, morphologically intermediate forms were reported. The present study aimed to identify these forms by the use of molecular characterization of DNA sequence. Based on morphometric criteria, adults of Fasciola hepatica, F. gigantica and intermediate forms were collected from naturally infected sheep and cattle from various regions of Sohag Governorate. A simple and rapid new method (QIAamp DNA Mini Kit) was used to isolate DNA from the worms and their RELP patterns were obtained after digestion of the PCR products with AvalI restriction enzymes. The result of a regular PCR experiment for the amplification of the selected 28S rDNA fragment with the designed primer set yielded identical 618- bp-long PCR products for the three types of Fasciola where the RFLP profile obtained from F. hepatica revealed four fragments of 628, 575, 165 and 95 bp, and F. gigantica generated three fragments corresponding to 628, 358 and 300 bp fragments whereas the intermediate forms revealed four fragments of 628, 541, 358 and 300 bp, which were similar to those ofF. gigantica but with a distinctive fragment of 541. These results confirmed that three species are present in our locality: F. hepatica, F. gigantica and an intermediate form which was named F. hepatogigantica n.sp. on basis of having few morphometric characters from F. hepatica (length and pattern of uterine coils) but genetically they were more related to F. gigantica.

Ectopic fascioliasis (EF) has direct and indirect effects on both humans and animals. The phenomenon of EF was individual cases in the period from 1950 up to the end of last century. From the period of 2000 up to 2006, plenty of researches were on EF in the developed and undeveloped countries. Nineteen EF cases infected with the immature and few with the mature worms were 13 females and 6 males. Three cases of lymphatic, pleural and breast fascioliasis reached the adults and laid their eggs in a lymph node in the cervical region pleural cavity and breast tissues. Until recent, knowledge about the ectopic fascioliasis pathway is little. Fasciola hepatica was the commonest species in most cases. The effect of fascioliasis might be direct to liver as ectopic foci or indirect on other organs due to the metabolites and secretory excretory products. All ages and both sexes were EF infected. Watercress topped the list of water plants born encysted metacercariae followed by lettuce, mint, and alfalfa. Nearly 24 million Egyptians at risk and about 800,000 were infected. On the global scale, about 180 million are at risk of infection.

Experimental infections of Egyptian Radix natalensis and French Galba truncatula with miracidia of Fasciola gigantica of sheep were carried out to determine the larval productivity of this parasite. Rediae and cercariae were thus counted in snails dissected at regular intervals from day 21 to day 49 postexposure (p.e.) at 24 degrees C, while cercarial shedding was studied in other two groups of snails after day 30 p.e. At day 49, the total number of free rediae and that of cercariae-containing rediae in R. natalensis (shell height, 10.0 mm) were 71.5 and 44, respectively, whereas mean values in G. truncatula (shell height, 5.7 mm) were 57.3 and 33 rediae, respectively. The life span of cercaria-shedding snails, the prepatent period, the patent period, and the total number of cercariae shed showed insignificant differences between both snail species. Compared to the data already obtained with a cattle isolate of parasite, the number of live rediae was significantly greater in G. truncatula and significantly lower in R. natalensis when exposed to sheep-originating miracidia. In cercariae, the differences between cattle- and sheep-derived infections were insignificant, whatever snail species. The results may be explained by the existence of an interpopulation of snail infection with F. gigantica, probably due to variations in frequency of natural encounters between this snail population and the parasite isolate. However, the better production of rediae and cercariae in G. truncatula might be due to the origin of snails used for this study because allopatric snails produced more larvae than sympatric congeners when they are subjected to experimental infections.

Fascioliasis is one of the familiar zoonotic health problems of worldwide distribution including Egypt. In this study, a simple and rapid polymerase chain reaction/restriction fragment length polymorphisms (PCR/RFLPs) assay, using the common restriction endonucleases Aval, EcoRI, Eael, Sac11 and Avail was applied to differentiate between both Fasciola gigantica and F. hepatica. The five restriction endonucleases were used to differentiate between the two species of Fasciola based on -1950 bp long sequence of the 18S nuclear small subunit ribosomal RNA gene. Aval and EcoRI restriction endonucleases failed to differentiate between the two Fasciola species when each restriction enzyme gave the same restriction patterns in both of them. However, F. gigantica and F. hepatica were well-differentiated when their small subunit ribosomal DNA were digested with Eael and Sac 11 restriction endonucleases.

In Egypt, zoonotic fascioliasis is increasing. In this study, postmortum examination of 88 donkeys used as gargantuan meal in the Zoo at Giza revealed hepatic fascioliasis in 15 (17.05%). The serum examination for anti-Fasciola antibodies by ELISA showed positivity in 12/15 with crude worm antigen, and positivity in 14/15 with locally prepared Fasciola excretory-secretory (Fges) antigen. The zoonotic role of animal fascioliasis was discussed.

Scanning electron microscopy (SEM) of adult Fasciola gigantica, a known liver fluke in Egypt, shows the presence of tegumental spines and folds giving the appearance of a rough surface. The spines are small and closely-spaced anteriorly, increasing in size and number on the in the mid-lateral aspect of the ventral surface. Towards the posterior end the spines progressively decrease in both size and number. Tegumental transverse folds or invaginations are interspaced between the spines. There are two types of sensory papillae on the tegumental surface. The dorsal surface exhibits similar features but the spines and papillae are less numerous with less extensive surface folds than on the ventral surface of the body. The oral and ventral suckers, together with the excretory pore, appear spineless, bearing ciliate sensory papillae. Further studies should be directed at deciphering the genetic codes and the synthesis of some of these antigens by recombinant DNA technology to be used in serodiagnosis, vaccination and for drug receptors.

The study assessed changes in fasting serum gastrin level in early schistosomiasis and chronic fascioliasis in 52 patients. Forty patients had either schistosomiasis or fascioliasis alone. Twelve patients had combined infection with both parasitosis. Also, ten healthy parasite free individuals were selected as a control. All were clinically examined and subjected to stool analysis, liver function tests and sero-diagnosis for H. pylori. Serum gastrin was measured by radioimmunoassay and showed significant elevation in all patients (with schistosomiasis, fascioliasis or both parasites) compared to controls. A significantly higher serum gastrin level was detected among patients with combined infection relative to either with schistosomiasis or fascioliasis. Alkaline phosphatase activity increased significantly in all patients as compared to control. The activity of the enzyme had significant positive correlation with egg counts in patients with either parasite and with serum gastrin level in those with combined infection. The results were discussed.

A total of 213 farm animals in the vicinity of Al-Santa Center (73 cattle, 90 buffaloes and 50 sheep) were coprologic examined for natural infection with trematod-parasites. The results showed that cattle were infected with Fasciola sp (21.8%) and Paramphistomum sp. (7.3%), buffaloes were infected with Fasciola sp. (17.7%) and Paramphistomum sp. (10%), while sheep were infected with Fasciola sp. (30%), D. dendriticum (5%) and Paramphistomum sp. (4%). The three animal species were treated for paramphistomiasis with a total dose of 1800, 6000 and 7500 mgm of Oleo-resin solution of Commiphora molmol (dose of 6 ml of 10gm% equal to 2 Mirazid. The cure was 100% in sheep 80% in cattle and 44.4% in buffaloes. High dose for both cattle and buffaloes to reach 100% cure rate was not tried.

Eight-hundred-twelve local and imported sheep, slaughtered at Riyadh abattoir, were subjected for parasitological diagnosis of fascioliasis by detection of eggs in the stool and worms in the liver and for serological diagnosis by detection of circulating anti-Fasciola IgG (CAFIgG) and circulating Fasciola antigens (CFAgs) using the indirect ELISA and the double antibody sandwich ELISA respectively. Detection of eggs revealed 13.5% infection rate compared with 21.9% by detection of worms (p<0.001). Infection rate was significantly higher (p<0.001) among the imported sheep (15.1%) than among the local ones (4.96%). Positivity for CAFIgG was 64.3% among all sheep while that of CFAgs was 30.2% (p<0.001). Sheep passing Fasciola eggs showed positivity of 99.1% for CAFIgG and 87.3% for CFAgs (p<0.05). Sheep free from any parasitic eggs showed positivity of 61.2% for CAFIgG and 21.1% for CFAgs. There was a highly significant association (p<0.001) between positivity of both CAFIgG and CFAgs and positivity of eggs in the stool.

Animal fascioliasis has been reported in Saudi Arabia among imported and local sheep. The paper demonstrated the parasitological and clinical features of human fascioliasis in nine out of ten male immigrant manual workers with manifestations suggesting fascioliasis. The sedimentation and Kato-Katz techniques proved effect in diagnosing Fasciola species eggs in human stool. The common clinical features were abdominal distension, flatulence, tender right-upper quadrant and easy fatigability and the least was the tinge of jaundice. Others as right upper quadrant pains, colicky abdominal pains & vomiting, epi-gastric pain and mild fever, and tympanitic abdomen were encountered. Anaemia and eosinophlia were also encountered in the ten patients. Fascioliasis patients (nine) were successfully treated with Mirazid as two capsules (600 mg) on an empty stomach an hour before breakfast for six consecutive days. Follow-up clinically and parasitologically was available in only seven fascioliasis patients who were completely cured. Follow-up for the other two fascioliasis patients was out in hand. Other parasites recovered in the stained (eosin, iodine and Zeihl-Nelson stains) smear stool samples was Entamoeba histolytica, Giardia lamblia and Cryptosporidium parvum. Besides, three were free from intestinal protozoan. The results were discussed on the light of the other work carried out regionally.

Trichinella spiralis /Fasciola gigantica cross-reactive fraction was purified from T. spiralis larval extract by affinity column chromatography in which CNBr-Sepharose 4B was coupled with F. gigantica antibodies. The fraction consisted of six polypeptides of 191KDa, 178KDa, 149KDa, 106KDa, 101KDa and 32 KDa as revealed by SDS-PAGE. Analysis of the free amino acids of the fraction revealed 17 amino acids with high proportions of tyrosine and glutamic. Immunization of rabbits subcutaneously with the cross-reactive fraction in Freund's adjuvant followed by challenge with F. gigantica metacercariae resulted in reduction in worm burdens reached to 47.8%. While immunization of rats with the same fraction in Freund's adjuvant followed by infection with T. spiralis larvae resulted in reduction in worm count reached to 74%. IgG antibody response in rabbits increased due to immunization to reach its maximum value at the time of infection and then decreased gradually up to the end of the experiments. But. remained higher than the level in non vaccinated control animals. In rat sera, IgG level increased due to vaccination but the level recorded its optimum value one week post infection and then decreased. Thus the cross-reactive antigen proved cross-protection with the protection inducing capability against both diseases.

In the present work, a total of 4180 sheep were obtained from different localities at Dakahlia Governorate. One hundred and eighty male sheep were investigated for Fasciola worms (the adult and juvenile forms) at Mansoura governmental abattoir. Four hundred grazing sheep were screened by the coprological examination from ten centers. The mean number of eggs per gm in a group of ten sheep ranged from 2.9-10.8 egg/gm. The number of worms ranged from 5-84. In the Mansoura abattoir, it was found that 37 sheep were infected (20.56%). Fasciola worms were recovered from the livers in 100%, common bile ducts in 60%, gall bladder in 80%, two sites in 50% and three sites in 40% of infected biliary tracts. According to the coprological examination, the infection rate was the highest in El-Manzalla Center (15.75) and the lowest was in Mansoura Center (3.25%). According to the prevalence and egg output results, it is concluded that sheep are the main reservoir host for fascioliasis among the edible animals in Dakahlia Governorate. However, examination of the non-edible animals and the domestic and wild rodents are ongoing and will be published soon elsewhere.

A field survey was done in Ezbet El-Bakly, Al-Fayoum Governorate, Egypt to assess the efficacy and safety of Mirazid in the treatment of human fascioliasis. Among 1019 individuals examined for parasitosis, the prevalence of fascioliasis was 1.7% and the geometric mean egg count (GMEC) was 33.2 eggs/gram stools. About 23.5% of the fascioliasis patients were asymptomatic. The most frequent symptoms were abdominal dis\tension and flatulence (76.5%), right hypochondrial pain (17.6%) and epigastric pain (17.6%). The most prevalent signs were pallor (52.9%,), tender right hypochondrium (23.5%) and tinge of jaundice (17.6%). All cases were treated by Mirazid as two capsules (600 mg) on an empty stomach an hour before breakfast for six consecutive days and followed up clinically and parasitologically. The parasitological cure rate, two and three months after treatment was 88.2% and 94.1% with an overt clinical cure without any side-effects. The cases not completely responding to a single course of treatment showed a marked reduction of the egg intensity. It was concluded that Mirazid (or Myrrh extract of Commiphora molmol) is safe and effective in the treatment of human fascioliasis under the field conditions.

A total of 21 children with fascioliasis (8 males and 13 females) with mean age of 10.4 years, 8 children with schistosomiasis mansoni (6 males and 2 females) with mean age of 11.37 years were treated with Myrrh (Mirazid) which is an oleo-gum resin from the stem of Commiphora molmol tree (Family Burseraceae). Also, ten healthly cross matched children were utilized as controls. Diagnosis was based on the detection of Fasciola hepatica or Schistosoma mansoni eggs in stool by Kato-Katz technique. Mirazid was given as 10 mg/kg/d an hour before breakfast for 3 consecutive days in schistosomiasis and for 6 days in fascioliasis. Clinical evaluation and stool analysis were done initially and at 2, 4 and 12 weeks post treatment to evaluate cure. Rectal snip was done for responding schistosomiasis cases to confirm recovery. Automated complete blood count with manual assessment of eosinophils, serum total IgE (enzyme immunoassay) and in vitro cytokines assay (IL-1 beta, IL-4, IL-5) by ELISA were performed for all subjects before treatment and repeated 12 weeks only for patients after therapy. Parasitologic cure was 90.9% in fascioliasis and 100% in schistosomiasis at 4 weeks post treatment. After a second dose Fasciola patients who remained positive were cured. Total IgE was significantly higher in Fasciola and Schistosoma patients before treatment compared to control (p < 0.001; 0.005 respectively) and decreased significantly with therapy (p = 0.001; 0.036). IL-1beta was higher in both patient groups than control (p < 0.001; 0.003) and decreased significantly 12 weeks after therapy to control level (p < 0.001; 0.017). IL-5 was high before treatment in both groups (p = 0.041; 0.027) and decreased significantly after 12 weeks after therapy (p = 0.005; 0.012). IL-4 did not differ from control before therapy (p = 0.58; 0.79) but increased significantly after treatment in both patient groups (p = 0.04; 0.02). It is concluded that Mirazid is an effective fasciolicidal and schistosomicidal drug. IL-1beta and IL5 were high in fascioliasis and schistosomiasis, but decreased with therapy denoting immunopathogenesis. The depressed IL-4 production may be a parasite immune evasion or host regulatory mechanism and cytokines levels may be criteria of cure.

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.

Fasciola hepatica whole worm homogenate (Fhwwh) separated fractions were used in enzyme linked immunoelectrotransfer blot (EITB) to identify the antigen(s) which induces antibody formation in human fascioliasis. The immuno-reactive antigens recognized by the infected patients were 25-29 kDa and 12 kDa. Antigens were biochemically purified by model 491-prep cell fraction (BIO-RAD). The capability of the purified target antigens to induce humoral and cellular responses with cells and sera of infected patients was investigated using enzyme linked immunosorbent assay (ELISA) and lymphoproliferative responses techniques. The 25-29 kDa cluster of antigen(s) were found to be more efficient in inducing lymphoproliferative response than 12 KDa, thus, it was considered as the target antigens used in the generation of human monospecific polyclonal immunopurified antibody probes (IPAb). The specificity and immunoreactivity of the IPAb with Fhwwh, F. hepatica excretory secretory products (FhESP) and S. mansoni adult worm antigen (SAWA) were evaluated by using EITB. Results showed that IPAb was immunoreactive with 25-29 and 12 kDa antigens of both Fhwwh and FhESP. It was concluded that the most immunogenic F. hepatica target antigen(s) were 25-29 and12 KDa antigens and there is a cross-reactivity between 25-29 and 12 KDa antigens in both Fhwwh and FhESP. The involvement of IPAb in antibody dependent cell mediated cytotoxicity in-vitro technique was studied. Results indicated that human neturophils were more effective in adhering to the IPAb-coated flukes at early stages of development.

Stool examination using modified Kato thick smear method was performed to detect Fasciola eggs and other parasites. Abdominal pain was the major presenting symptom (87.7%) followed by pallor (83.3%) and fever (16.7%). Anaemia and hepatomegaly were recorded in 77.7% of patients compared to 27.7% with splenomegaly. Abdominal ultrasonography revealed hepatomegaly and common bile duct dilatation in 77.7% of patients. Moreover, 5 cases showed diagnostic. Olympic game rings All patients had positive IgG4 levels, 55 cases were positive for specific total IgG and IgG1, whereas, only 24 cases had positive IgG2 levels (26.6%). All negative control group showed no cross reactions. On the other hand, ELISA detecting IgG4 showed the highest specificity (95%), followed by IgG2 (85%) and the least specific test was obtained with detection of IgG (70%) and IgG1 (65%). One month after treatment, 91.1% of patients (82/90) were completely cured and even after another two months follow-up. In completely cured patients none of anti-Fasciola isotypes was significantly changed. So, detection of anti-Fasciola isotypes especially IgG4 is very specific for the accurate diagnosis of human fascioliasis.

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.

Examination of twenty parasitologically proven human cases of fascioliasis showed the following clinical, laboratory and ultrasonography results: (1) Both sexes are susceptible to fascioliasis. The smallest one was a child 3-year old. (2) The most important clinical manifestations are: a- abdominal distention and flatulence (30%), b- right upper quadrant pains (25%), colicky abdominal pains (25%), d- pallor (25%), and etympanitic abdomen (25%). Nevertheless, 15% of the patients showed no symptoms. (3) The most significant items in the CBC and liver function tests are: a- significantly high eosinophilia, b-high alkaline phosphatase, and c- low haemoglobin. Besides, two fascioliasis patients had high serum bilirubin. (4) Ultrasonography diagnosed only nine fascioliasis patients (45%). The result was critically discussed.

No doubt, human fascioliasis is an increasing worldwide zoonotic liver fluke. Clinically, human fascioliasis has to be differentially diagnosed from many hepatic diseases as acute & chronic hepatitis, schistosomiasis mansoni, visceral toxocariasis, visceral leishmaniasis, hepatic amoebiasis, biliary tract diseases and others. The parasitological diagnosis based on the demonstration of the eggs in stool, duodenal contents or bile is usually unsatisfactory due to false passage of eggs, ectopic fascioliasis, and failure of immature worm to maturation. So, ELISA-Fhes antigen (Fasciola hepatica excretory/secretory) and IHAT were evaluated in the immunodiagnosis of parasitologically proven cases of human fascioliasis compared with proven cases of human schistosomiasis mansoni and parasite-free individuals. ELISA-Fhes gave 100% sensitivity and 100% specificity. On the other hand, IHAT was less sensitive and less specific.

Fascioliasis, caused by Fasciola species, is a disease of herbivorous animals. It has a worldwide distribution in a large variety of grass-grazing animals as sheep, goats, cattle, buffaloes, horses and rabbits. In Egypt, donkeys and camels as well, are hosts for F. gigantica. Fascioliasis may occasionally affect man. Human infection causes serious hepatic pathological sequences that add to the already known threats to the liver of the Egyptian population. Two clinical stages are recognized in human fascioliasis. An acute stage coincides with the larval migration and worm maturation in the hepatic tissue, and a chronic stage coincides with the persistence of Fasciola worms in the bile ducts. Human infection with fascioliasis was very sporadic until the last three decades where clinical cases and outbreaks were reported. The estimated the number of people currently having fascioliasis to be 360,000 in Bolivia, 20,000 in Ecuador, 830,000 in Egypt, 10,000 in Islamic Republic of Iran, 742,000 in Peru, and 37,000 in Yemen. The total estimated number of people infected is 2.4 million in 61 countries and that the number at risk is more than 180 million throughout the world. Human fascioliasis has to be differentially diagnosed from some diseases as acute hepatitis, infection with other liver flukes as schistosomiasis, visceral toxocariasis, biliary tract diseases and hepatic amoebiasis. The parasitological diagnosis is based on identification of eggs in stool, duodenal contents or bile, also by the recovery of adult worm during surgical exploration, after treatment or at autopsy. However, the eggs may be present in very small number at irregular intervals, hence difficult to be found. Besides, the eggs may be transiently present in stool after ingestion of raw or undercooked liver from infected animals. The direct methods of diagnosing the egg are usually unsatisfactory. The symptoms may be present for several weeks before eggs are recovered in stool. Thus, the serologic tests are the alternative method of confirming early and extrabiliary human fascioliasis. However, cross-reactions with other helminthic antigen may confuse the interpretation of the results.

Seventeen patients with established fasciolosis and ten normal controls were enrolled in the study. The Fasciola patients were divided according to infection intensity into two groups (four patients with high intensity and thirteen patients with low intensity) as assessed by egg counts coupled with ultrasonography for detection of worms in the biliary system. Aspartate and alanine aminotransferases (AST and ALT) levels were similar to those of the controls, within the accepted normal limits, before and after treatment denoting absence of hepatocellular injury. Total serum bile acids, individual bile acids: cholic acid (CA) and chenodeoxycholic acid (CDCA), gamma glutamyl transpeptidase (GGT) and serum alkaline phosphatase (SAP) were significantly higher among all patients as compared to the controls denoting a degree of cholestatic lesion in those patients. Patients with high infection intensity revealed higher parameters than those with low intensity. The difference was not significant. One month after treatment, there was a significant improvement in the cholestasis indicating parameters in all Fasciola cases compared to the pretreatment ones. This indicates the effective role of the drug on the hepatobiliary function. However, the levels were still different from the controls. In Fasciola infection, total and individual serum bile acids in conjunction with GGT and SAP evaluate the hepatobiliary status and detect any minor abnormalities especially in anicteric subjects. Studied after treatment, they can be useful indices for assessment of the improvement.

Fascioliasis is now imposing itself as a zoonotic disease in sheep and cattle raising countries. In Egypt, human fascioliasis is increasing. During the years 1994 to 1997 the overall slaughtered animals in Egyptian abattoirs was 2,003,200 sheep and goats, 2,624,239 cattle and 3,536,744 buffaloes. The overall rates of fascioliasis were 2.02% for sheep and goats, 3.54% for cattle and 1.58% for buffaloes. Macroscopic examination of sheep liver showed up to 100 flukes per liver inside a largely dilated thick walled bile ducts. Cattle liver showed up to 275 flukes per liver inside thickened dilated and calcareous bile ducts with offensive yellowish brown bile. Buffaloes liver showed up to 330 flukes per liver. Microscopic examination showed mainly thickened wall, hyperplasia and marked fibrosis. The discussion focused on the zoonotic importance of fascioliasis in Egypt, diagnosis, treatment and prevention. No doubt, adequate control of fascioliasis is more or less a problem, since it requires the control of snail intermediate host(s) and control of infection in all affected animals and man.

The objective of the present work is to evaluate Fasciola E/S antigens for diagnosis of early fascioliasis utilizing different diagnostic techniques. Using enzyme-linked immunoelectro-transfer blot (EITB), Fasciola-specific E/ S protein fraction band (49.5 kDa) was determined and electroeluted. The mono-specific antibodies against this specific fraction band were prepared by immunizing pathogen-free rabbit. Assessment of the prepared mono-specific antibodies in diagnosis of human fascioliasis was performed through the detection of E/S copro-antigens by enzyme-linked immunosorbent assay (ELISA) in stool eluates obtained from patients with confirmed fascioliasis, other parasites as well as from other healthy individuals. Serum samples were collected and tested to detect serum antibodies against Fasciola E/S antigen using EITB and counter immunoelectrophoresis (CIEP). Analysis of Fasciola adult worm E/S products by SDS/PAGE revealed a number of bands, the molecular weight (MW) of which ranged from 14-200 kDa; with three major bands (27.5, 32.5 and 55 kDa). Fasciola EIS 49.5 kDa protein fraction proved to be specific to F. gigantica. Cross reaction with S. mansoni was observed at higher MW (110-120 kDa). The sensitivity, specificity and diagnostic accuracy of EITB were 45.2%, 100% and 70.7%, respectively, while those of CIEP were 38.7%, 100% and 67.2%, respectively. ELISA technique using mono-specific anti-49.5 kDa to detect copro-antigens proved to be practical and reliable. It showed higher sensitivity (91.4%) and higher diagnostic accuracy (91.8%), while the specificity was 92.3%. In addition, ELISA had higher negative predictive value (88.9%) and fair positive predictive value (94.1%).

The levels of superoxide (O(-)2) and nitric oxide (NO) production by monocytes have been measured in 15 patients with S. mansoni, 15 patients with Fasciola and six patients with combined infection as well as in control group (15). The levels of both radicals were significantly higher in all patient groups than in the control group, indicating that these radicals may have a role in the immunity against such infections. Patients with chronic fascioliasis showed lower level of O(-)2 and NO than those with schistosomiasis. This may be due to the lodging of the mature Fasciola spp. away from the immune system and subsequently decreased amount of antigens reaching the circulation. In combined infection, the levels of these products were at the highest value, due to increased antigenic stimulation and cross reactivity between the two parasites which may have lead to augmented immune response.

The production of pro-inflammatory cytokines by monocytes in vitro has been measured in eight patients with acute fasciolosis and 15 patients in the chronic stage of the disease, before and after stimulation by excretory/secretory Fasciola antigen. Results were compared with those of a control group of 12 individuals. The monocytes from patients with acute fasciolosis produced significantly higher levels of GM-CSF, IL-8 and IL-6 as compared to controls. With chronicity, the production of these cytokines was decreased as compared to the acute stage probably due to decreased antigen level in blood. Stimulation of monocytes of healthy control with E/S Fasciola antigen was accompanied with a markedly increased production of pro-inflammatory cytokines, while monocytes from patients with acute or chronic fasciolosis revealed minimal increase in production. This denoted the importance of E/S Fasciola antigen as an activator of monocytes. A second exposure to the same antigen was accompanied with a limited response.

A comparative scanning electron microscopy and morphometrical study of the two geographically isolated species of Fasciola (F. heptica European isolate and F. gigantica from Egypt) were studied in order to clarify their genetic relationships and specific identification. Although the present study has revealed that most of the diagnostic morphological and morphometrical criteria in the two species are highly variable, the position of the ventral sucker relative to the whole body length produced the most significant differentiating criterion, in addition to the presence of markedly larger tegumental papillae on the ventral surface of F. gigiantica.

An immunodiagnostic sandwich enzyme-linked immunosorbent assay (ELISA) was performed for the detection of F. hepatica antigens in stool sample (copro-antigens) of experimentally infected animals (mice, rats and rabbits) over a period of 18 weeks using affinity-purified polyclonal antibodies raised against the whole adult F. gigantica. The test demonstrated fascioliasis in mice earlier than in rats or rabbits (4, 6 and 7 weeks post infection respectively) by detection of copro-antigens in stool samples. In all experimentally infected models, F. hepatica copro-antigens concentration showed continuous increase and mice samples had higher antigen concentration than rats and rabbits. Sensitivity of ELISA was significantly higher in mice (83.3%) than in rats and rabbits (75% and 66.7% respectively) and its specificity showed nearly similar results to sensitivity. From the previously obtained data, it could be concluded that ELISA proved to be a rapid, easy and sensitive test for diagnosing fascioliasis by detection of F. hepatica copro-antigens earlier than routine stool examination of Fasciola eggs.

Rabbits were infected with 30 metacercariae each. They were then divided into four batches each of two groups. One batch served as controls and the others were treated each with a drug. The first group from each batch was treated when worms were still immature, the other group was treated after maturity. The effect of the drugs was monitored by: (1) Stool examination for eggs and studying the percent of egg reduction. (2) Worm recovery at autopsy and calculating the percent of the drug efficacy. (3) Culturing some eggs and following their embryonation and hatching. Results pointed out that Triclabendazole was the best fasciolicide followed by Bithionol. Praziquantel had a negligible effect.