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[Therapeutic alternatives in case of failure of first-line treatment of intestinal helminthiasis in adults].

Development of antiparasite medications over the last 15 years has greatly reduced the number of treatment failures for intestinal helminthiasis. Benzimidazole derivatives, ivermectine, praziquantel and triclabendazole are easy to use, well tolerated and generally curative. First-line treatment are currently so reliable that failure should lead first to investigation of possible "false failure" causes such as misdiagnosis, poor identification of the parasite, inadequate or incorrect treatment, and repeat contamination, before concluding that genuine parasite resistance is involved and that alternative therapy is needed. Nitazoxanide is an alternative treatment for fascioliasis and teniasis. Albendazole can be beneficial for taeniasis and strongyloidiasis. Metronidazole can be effective for fascioliasis. Artemisinine derivatives are useful for schistosomiasis. Combined therapies are necessary for refractory ankylostomiasis.

Albendazole↗

Endoscopic removal of one live biliary Fasciola gigantica.

Biliary fascioliasis was diagnosed in a woman from Angthong province, who presented herself with anorexia, weight loss, and jaundice for one month. Intraoperative retrograded cholangiopancreatography revealed a filling defect considered as a stone and bile sludge. By the sphincterotomy and balloon-extraction technique, one live fluke was removed and identified as Fasciola gigantica. A single dose of praziquantel, 25 mg/kg of body weight/day, was given although the postoperative stool examination revealed no Fasciola spp. egg. The patient was doing well after the fluke removal. According to a previous review, in this country, Ayutthaya is the southernmost province for the distribution of fascioliasis.

Aged↗

Cross-reactions in immunodiagnosis of patients infected with Schistosoma, Fasciola and Heterophyes using ELISA.

Cross-reactions in immunodiagnosis using schistosomal soluble egg antigen (SEA) and Fasciola adult worm antigen in cases with fascioliasis, heterophyiasis and schistosomiasis were studied by ELISA techniques. False positive reactions occurred in 10% of cases with fascioliasis and heterophyiasis respectively when using schistosomal SEA. Using Fasciola crude antigen, false positive reaction occurred in 25% of cases with schistosomiasis and 35% with heterophyiasis. The specificity of SEA in heterologous cases was 87% while that of Fasciola antigen was 70%. This difference was found to be statistically significant.

Animals↗

Acute cholecystitis and Fasciola sp. infection in Thailand: report of two cases.

Two cases of fascioliasis gigantica from Northeast Thailand presenting with cholecystitis and gall stones were reported. Both cases complained of abdominal pain. On laparotomy the worms were found. In one case five worms were recovered during bile duct exploration and bile drainage; eggs were also revealed in the bile. In the other case of ectopic fascioliasis one young adult worm was found in a nodule which adhered to liver and diaphragm. The parasites were identified as Fasciola gigantica.

Abdomen, Acute↗

Antibody profiles by EITB and ELISA of cattle and sheep infected with Fasciola hepatica.

In evaluating potential mechanisms of immunity in fascioliasis we compared the time-course analysis of the antibody responses between a resistant (cattle) and a susceptible model (sheep). Sera from sheep and cows experimentally infected with F. hepatica were reacted with both somatic (FhWWE) and excretory-secretory (ES) antigens in order to evaluate the antibody repertoires in the 2 different hosts. Analysis of these sera by ELISA showed a significant increase in antibody levels by 2 wk in most infected cattle using both somatic and ES antigens, whereas with most infected sheep antibodies are not clearly detected until week 4. By EITB, both infected sheep and cows recognize major somatic polypeptides in a molecular weight range of 30-38 kDa by 8 wk. Cattle recognized 3 additional major antigens of 56, 64, and 69 kDa as early as 6 wk. Various polypeptides of 20-25 kDa are prominently detected by most sheep and very faintly, if at all, by the cow sera. The sera from both sheep and cows also identify ES polypeptides of 20-28 kDa. The patterns of polypeptides recognized by sheep infected with S. mansoni and challenged with F. hepatica in EITB are almost identical to those with a simple F. hepatica primary infection. No significant differences were detected in the antibody kinetics in ELISA between these 2 groups. Differences and similarities between these models could eventually help determine which antibodies may be predictive of resistance or susceptibility in fascioliasis.

Animals↗

Bleeding ulcer in the common bile duct due to Fasciola hepatica.

Fatal complications caused by Fasciola hepatica are very rare. We describe the case of a 5-yr-old girl who died of a rare complication of fascioliasis--ulceration of the common bile duct by the parasite, which provoked acute hematobilia. This is the first report of a case in which fascioliasis was presented clinically as upper gastrointestinal bleeding. Normally, this parasite hatches in the duodenum, pierces the intestinal wall, and enters in the liver where it finally reaches the bile ducts. In this case the parasite probably entered the papilla of Vater and continued into the common bile duct where it attempted to perforate the wall of the duct, repeating the normal process used to perforate the wall of the duodenum.

Child, Preschool↗

Comparison of adult somatic and excretory-secretory antigens in enzyme-linked immunosorbent assay for serodiagnosis of human infection with Fasciola gigantica.

Adult somatic antigen extract of Fasciola gigantica was compared with excretory-secretory (ES) antigen in an enzyme-linked immunosorbent assay (ELISA) for serodiagnosis of human fascioliasis gigantica. The absorbance values in ELISA using the adult somatic antigen were not significantly different from the values obtaining using ES antigen (p > 0.05). The diagnostic sensitivity, specificity and positive and negative predictive values of the test using adult somatic extract as antigen were 100%, 98%, 70% and 100%, respectively. On the other hand, these values of the test using adult ES antigen were 100%, 99.3%, 87.5% and 100%, respectively. It appears that both somatic and ES antigens are effective antigens for use in the serodiagnosis of human fascioliasis gigantica.

Animals↗

Role of ELISA in early detection of Fasciola copro-antigens in experimentally infected animals.

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.

Animals↗

A monoclonal antibody diagnoses active Fasciola infection in humans.

Monoclonal antibodies (MABs) were produced after fusion of spleen cells of Fasciola antigen immunized BALB/c mice and non secreting murine myloma cells (P3x63 Ag8). Six MAbs, showing the highest reactivity with purified Fasciola antigen, were prepared. All 6 MABs were IgG2 with Kappa light chain. Reactive epitopes recognized by the six MAbs were glycoprotein in nature, and each MAb recognized a single epitope of Fasciola antigen. No cros reactions were observed with Schistosomal AWSA, hydatid Ag and Entamoeba Ag. EITB technique showed a specific diagnostic band at 17.5 kDa for each of the six MAbs. Anti-Fasciola MAb (AD2) was conjugated with peroxidase and was used with anti-rabbit anti-Fasciola polyclonal antibody in sandwich-ELISA to detect circulating Fasciola antigen in serum and urine samples of 57 fascioliasis patients, 51 schistosomiasis patients, 45 patients infected with other parasites and 47 healthy controls. Sensitivity of the assay in detection of circulating Fasciola antigen in sera and urines of Fasciola infected patients was 100%. The specificity of the assay was calculated among patients infected with schistosomiasis and other parasites and was 98% in serum and 97% in urine. A positive correlation was found between levels of circulating Fasciola antigen in serum and urine samples of fascioliasis patients (r = 0.825, p < 0.05).

Adolescent↗

DNA probes for the detection of Fasciola hepatica in snails.

Fasciola hepatica, also called the large liver fluke, is a trematode which can infect most mammals. Monitoring the infection rate of snails, which function as intermediate hosts and harbour larval stages of F. hepatica, is an important component of epidemiological studies on fascioliasis. For this purpose, DNA probes were generated which can be used for the detection of F. hepatica larvae in snails. Four highly repetitive DNA fragments were cloned in a plasmid vector and tested by Southern blot hybridization to the DNA of various trematodes for specificity and sensitivity. The probes Fhr-I, Fhr-II and Fhr-III hybridized only to F. hepatica DNA. Fhr-IV contained ribosomal RNA gene sequences and cross-hybridize with the DNA from various other trematode species. Squash blot analysis showed that the different probes were able to detect the parasite larvae in trematode-infected snails even as isolated single larvae. No signals were obtained in squash blots of uninfected snails. Probes Fhr-I, Fhr-II and Fhr-III are thus useful specific tools for studying the epidemiology of fascioliasis. The probe Fhr-IV, because of its broader spectrum, can be used to detect the larvae of a wide range of trematode species of waterbirds, which are the causative agents of swimmer's itch.

Animals↗

Application of thin layer immunoassay (TIA) as a serodiagnostic tool in schistosomiasis. A preliminary report.

Thin layer immunoassay (TIA) is a recently developed simple serological technique which, in a preliminary study, has been applied to the serodiagnosis of schistosomiasis. 64 of a total of 69 sera from patients with schistosomiasis were positive in TIA when a Schistosoma mansoni worm antigen preparation was used as coating material. A trial of the diagnostic specificity of the assay was made by cross-testing sera from patients with different parasitic diseases, using TIA plates coated with extracts from the relevant parasites. All sera from patients with filariasis, fascioliasis and echinococcosis were positive in homologous TIA systems. In heterologous systems, cross-reactivity was noted for some sera in all groups except fascioliasis. The largest proportion of cross-reacting sera was registered in the echinococcosis group. It is suggested that, after further evaluation, the TIA technique might supplement or be used as an alternative to other serological tests already in use for the diagnosis of schistosomiasis. In favour of the TIA technique are its simplicity and its low cost.

Hemagglutination Tests↗

Revised post-mortem inspection procedures for cattle and pigs slaughtered at Australian abattoirs.

Revised procedures for the post-mortem inspection of cattle and pigs at Australian abattoirs were developed as minimum national requirements. Comparative trials of current and revised procedures were conducted at 3 and 4 Australian abattoirs for pigs and cattle respectively. Residual pathological changes after inspection and removal of lesions from heads, carcases or viscera were recorded by an evaluation team with the data used to determine relative effectiveness. For cattle, there was no significant difference between current and revised procedures. Although it had been proposed to eliminate routine incision of cervical lymph nodes for pigs, it was found necessary to retain this procedure. The data also revealed a significant variation in detection of pathology between abattoirs for both species. Parasitic conditions accounted for the greatest proportion of undetected lesions in both species for both current and revised procedures. These comprised mainly fascioliasis in cattle and ascariasis in pigs. The former was associated with the study being conducted in eastern Australia. Chronic interstitial nephritis was also a common undetected lesion, reinforcing the need for intensified inspection of kidneys. However, such residual pathology was considered to be of only limited significance to human health. The revised procedures and findings are discussed in the context of animal and human health and of the importance of the meat industry to Australian exports. Some attention has been given to specific conditions likely to be affected by the proposals including tuberculosis, bovine cysticercosis, fascioliasis, porcine arthritis and salmonellosis. The role of meat inspection in monitoring for specific diseases is discussed.(ABSTRACT TRUNCATED AT 250 WORDS)

Abattoirs↗

Identification of genes that encode Fasciola-specific arc 2 antigens.

Previous studies have shown that serum from humans with fascioliasis had precipitating antibodies that identified a line of precipitation as specific for Fasciola. This Fasciola-specific band was designated arc 2. The specificity of a Fasciola-specific anti-arc 2 antiserum, evaluated by Western blot analysis, reveals a mosaic of antigens with molecular weights in the range of 10-43 kD. In the current study, the antibodies reacting with a subset of antigens in the 10-21-kD range were eluted from nitrocellulose and used to probe an F. hepatica cDNA library. Four clones were purified and amplified by the polymerase chain reaction. The DNA hybridizations among these clones revealed different groups of clones derived from different mRNAs. The finding of multiple genes supports the observation that F. hepatica arc 2 is a mosaic of antigens. The studies show that molecular cloning technology can be used to identify potential genes, each of which encode distinct antigens that may be used for the specific immunodiagnosis of fascioliasis.

Animals↗

The food-borne trematode zoonoses of Vietnam.

During the past couple of decades, numerous surveys for the occurrence and distribution of food-borne trematodes in Vietnam have been carried out. However, the majority of the data obtained have not been published in international journals, and therefore, the seriousness of these trematode parasitic zoonoses in the country has not been fully appreciated. To correct this, over 40 Vietnamese language reports and local publications were translated and organized as a status review for an international audience. The results show that such serious trematode zoonoses as clonorchiasis, opisthorchiasis, paragonimiasis and fascioliasis are common in many regions of Vietnam, and, in the case of fascioliasis and paragonimiasis, are increasing. Data on the species of intermediate hosts involved in the transmission of these zoonoses, and the effect of host sex and age on infection frequencies are presented, along with findings on food preference/behavior investigations. Finally, the authors present recommendations for further research to provide a more comprehensive picture of the status of these zoonoses, and to obtain the risk assessment information needed to design prevention and control programs.

Adolescent↗

Cysticercosis: IgG-ELISA evaluations of peak1 antigen and <30 kDa antigen of delipidized extract of Taenia solium metacestodes.

The antigenicity of ether-delipidized Taenia solium metacestode extract (DLPAg) was investigated by IgG-ELISA. The antigen showed higher antigenicity than that of non-delipidized antigen (NDLPAg). Then the DLPAg was subjected to Sephacryl S-200 gel chromatography and a partially purified antigen (DLPP1Ag) was identified as the promised antigen by IgG-ELISA using 25 sera from cysticercosis cases, 177 cases of 24 heterologous infections, and healthy controls. Sensitivity was 52% and specificity was 91.8% at the cut-off value (X + 7SD), 0.399. Cross-reactivity occurred with 17 cases of eight diseases: cystic echinococcosis (7/11), taeniasis (1/16), gnathostomiasis (2/8), strongyloidiasis (1/12), angiostrongyliasis (1/12), paragonimiasis heterotremus (2/15), opisthorchiasis (1/9) and fascioliasis (2/7). When DLPP1Ag was fractionated through Ultra free centrifugal tube (retained 30 kDa) and Amicon (PM10), MWCOP1Ag (<30-10> kDa) was obtained; the antigen showed better results than DLPP1Ag with 88% sensitivity and 95.6% specificity at the cut-off value (X + 4SD), 0.264. Nine cases of six diseases cross-reacted with this antigen: cystic echinococcosis (2/11), gnathostomiasis (2/8), trichinellosis (2/12), toxocariasis (1/5), schistosomiasis (1/6), and fascioliasis (1/7). MWCOP1Ag gave higher sensitivity than that of DLPP1Ag but some cross-reactivity occurred.

Animals↗

Experimental infection of rabbits with Fasciola gigantica and treatment with triclabendazole (TCBZ) or BT6: parasitological, haematological and immunological findings.

The responses of rabbits experimentally infected orally with 25 Fasciola gigantica metacercariae each, to treatment with triclabendazole (TCBZ) in a single oral dose of 10 mg/Kg body weight or with the newly synthesized drug (BT6) in repeated oral doses of 500 mg/Kg for 3 consecutive days, were monitored by faecal egg and fluke counts and the enzyme-linked immunosorbent assay (ELISA), both treatments at 4-, 8- and 12- weeks of infection. In all infected rabbits, eggs appeared in the faeces between 10 and 16 weeks post-infection (P1) and Were egg-negative in the TCBZ-treated animals. Compared with the infected untreated control (group B), the reduction in the fluke burdens were 93.75,100 and 100% in TCBZ-treated rabbits (groups C, D and E), respectively and 33.33, 27.08 and 27.08% in the BT6 treated groups F, G and H, respectively. Total red blood corpuscle (RBCs) counts, haemoglobin (Hb) concentration and packed-cell volume (PCV) values in rabbits (group B) indicated the presence of moderate anaemia from the start of infection with a significant decrease from 10-12 weeks onwards. RBCs count, Hb and PCV values from groups C, D and E were significantly higher (p < 0.001) than in those from group B on week 10, 8 and 10 PI, respectively. The increasing was continued till week 16 PI. By two weeks PI there was significant eosinophilia in infected rabbits, maintained at 4 and 6 weeks PI then dropped up to 8 weeks but still statistically higher than the control values. The ELISA detected antibodies against F. gigantica as early as week 2 PI, rising to high levels 6, 10 and 12 weeks later in groups C, D and E, respectively, whilst peaked at 10 weeks in BT6 treated animals, then tended to drop but remained positive through 16 weeks. We conclude TCBZ is a potent fasciolicidic drug. Eosinophilia and anaemia might be indicators for fascioliasis. The ELISA with adult fluke excretory - secretory (ES) antigens could be a feasible method for the diagnosis of experimental fascioliasis in rabbits and post-treatment monitoring.

Animals↗

Indigenous livestock disease problems and slaughter cattle infections in the Ada district of the Ethiopian highlands.

In the farming enterprises of the Ada district, sheep and goats were most commonly diseased by pneumonia and helminthiasis among all farming animals, which led to severe losses of lambs and sheep. In cattle, helminthiasis and malnutrition conditioned considerable losses of calves. Faecal investigations showed that 72% of the cattle were infected with internal parasites, with nematodes more frequent in calves and fascioliasis among adult cattle. Pathological malformations of organs were encountered in 65% of the cattle slaughtered at the Debre Zeit abbatoir. Economically and hygienically most important are fascioliasis, hydatidosis, and cysticercosis.

Animals↗

Enzyme-linked immunosorbent assay using cysteine proteinase antigens for immunodiagnosis of human paragonimiasis.

An enzyme-linked immunsorbent assay (ELISA) using worm extract antigens from lung flukes of Paragonimus westermani provided good sensitivity to sera from patients with paragonimiasis westermani but high cross-reactivity with sera from most fascioliasis patients and some patients with onchocerciasis or clonorchiasis. To improve the specificity, we tested an ELISA using fluke cysteine proteinases as antigens. Cysteine proteinases were partially purified from the excretory/secretory products of P. westermani by 40-75% ammonium sulfate fractionation, hydrophobic chromatography, and arginine affinity chromatography. An ELISA using the enzyme preparation not only had increased sensitivity to paragonimiasis westermani sera but also reduced cross-reactivity with the fascioliasis, onchocerciasis, and clonorchiasis sera to negligible levels. The reactivity of the ELISA to paragonimiasis miyazakii sera was similar to that of paragonimiasis westermani sera. A proteinase preparation from P. ohirai, which can be obtained easily from infected rats, provided similar results. Therefore, the ELISA using cysteine proteinases of Paragonimus could not distinguish the parasite species with which patients were infected, but it is a valuable assay with which to immunodiagnose paragonimiasis even when the proteinases are prepared from nonhuman species.

Animals↗