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Update on Hepatobiliary and Pulmonary Flukes.

Hepatobiliary and pulmonary trematodes continue to cause significant morbidity and mortality worldwide. This article provides a selected update on the clinical features, diagnosis, immunology, treatment, and control of infections caused by Clonorchis sinensis, Opisthorchis species, Fasciola hepatica, and Paragonimus species. Although recent molecular studies have elucidated novel mechanisms of immune evasion in Paragonimus westermani and F. hepatica, there are no immediate prospects for a vaccine for any of these trematodes. Although treatment of paragonimiasis, clonorchiasis, and opisthorchiasis was simplified with praziquantel in the 1980s, eradication of fascioliasis remains difficult because of the limited availablity of bithionol. Several experimental drugs have been tested as possible alternatives. Despite successful pilot public health programs designed to decrease transmission of these trematodes, their prevalence remains high and underlines the importance of allocating resources to control them.

Journal Article↗

Trichinella spiralis: antigenic substances associated with the alimentary tract.

Postembedding immunogold and immunoperoxidase staining methods revealed that substances occupying the lumen of the esophagus and the midgut were antigenic for both Wistar rats and humans. Specificity of the alimentary tract-associated antigen was assessed by reacting these substances with a panel of serum pools from patients with non-Trichinella helminth infections, including anisakiasis, paragonimiasis, gnathostomiasis, fascioliasis, dirofilariasis, and trichuriasis. The substances had no, or negligible, cross-reactivity among the serum pools with the only exception of severe trichuriasis serum. Cytochemical staining revealed that the substances are PAS positive and are stained red by azan, and the midgut-occupying substance was equipped with exposed concanavalin A-binding sites. The present data suggest the alimentary tract-associated antigen can be purified by lectin affinity chromatography and may be used as a fairly specific antigen for immunodiagnostic purposes.

Animals↗

A survey and review of hepatobiliary lesions in Australian macropods.

This report presents information on the range of diseases and lesions that occurred in sections of livers of macropods held in the Non-Domestic Animal Registry at Taronga Zoo. Of 142 affected livers, 52 were due to parasites, 24 to bacteria, nine to Macropod Herpesvirus, four to fungal agents and ten to tumours. In addition, 17 livers had acute degenerative or necrotic lesions, 22 were affected by fibrosis or other chronic lesions and four had miscellaneous degenerative lesions. Common parasitic diseases included toxoplasmosis, coccidial cholangitis, cestode cholangitis (Progamotaenia sp.), fascioliasis and capillarial hepatitis. Bacterial diseases were varied but lesions due to anaerobic bacteria or Mycobacteria spp. were most common. Many of the acute degenerative lesions were associated with cardio-respiratory disease. Chronic lesions were often of unknown origin, although it was speculated that parasitic and bacterial organisms contributed to the aetiology. There were four primary and six metastatic tumours.

Animals↗

A paper radioimmunosorbent test (PRIST) for the detection of larva-specific antibodies to Toxocara canis in human sera.

A paper radioimmunosorbent test (PRIST) was shown to be sensitive and reproducible when used with excretory/secretory antigen of Toxocara canis second stage larvae. Whatman No. 50 filter paper (5 mm discs) gave the most consistent and clear results with antigen at a concentration of 100 micrograms/ml, and could be stored for up to 3 weeks in vacuo at -70 degrees C. Antigen coated discs were incubated with test sera at 1:10 dilution for 3 h at room temperature (21 degrees C), reacted with [125I]anti-human IgG for 1 h and counts determined in a gamma counter. Sera from patients with fascioliasis, taeniasis, schistosomiasis, oxyuriasis, trichinellosis and ancyclostomiasis gave counts similar to cord serum controls. Sera from patients with ascariasis gave counts of up to twice as great as controls, but sera from patients with toxicariasis produced counts of 7,000-13,000, a 4-6-fold increase.

Animals↗

Evaluation of total serum bile acid concentrations for the diagnosis of hepatobiliary disease in cattle.

Serum bile acid concentrations were measured in 41 clinically healthy cattle of different breeds. There was no diurnal variation in values and age and sex had no effect. There was no significant difference between serum and plasma bile acid concentrations in clinically healthy cattle. Serum bile acids were stable on storage at -20 degrees C. The total serum bile acid concentrations, together with other tests of hepatic disease, were evaluated in cattle with various types of hepatobiliary disease (hepatic lipidosis, hepatic abscessation, leptospirosis, biliary calculi, fascioliasis), respiratory, cardiovascular and infectious diseases, and in various other conditions not affecting the liver. Total serum bile acids were the most specific and sensitive indicators of a wide variety of hepatic diseases and were significantly correlated with the degree of clinical illness.

Animals↗

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

Adult somatic antigen extract of Paragonimus heterotremus was compared with excretory-secretory (ES) antigen in an enzyme-linked immunosorbent assay (ELISA) for serodiagnosis of human paragonimiasis. The absorbence values in ELISA using the adult somatic antigen were not significantly different from the values obtained using ES antigens (P greater than 0.05). The sensitivity of the ELISA using either antigen was 100%, but the specificity was 96% with somatic and 98% with ES antigens due to a cross reaction with fascioliasis sera. It appears that both somatic and ES antigens are effective antigens for use in serodiagnosis of human paragonimiasis heterotremus.

Animals↗

Isolation and purification of a diagnostic antigen for bovine cysticercosis by hydrophobic chromatography.

An ammonium sulfate fraction of Taenia hydatigena cyst fluid (ThFAS) was further fractionated by hydrophobic interaction chromatography, using alkylagarose and omega-amino alkylagarose columns, in an effort to isolate and purify a specific diagnostic antigen in the ThFAS preparation. The less than 12 kDa antigen was found to have an affinity for immobilized alkanes with chain length of six carbons or greater. The antigen was recovered in an ethylene glycol eluate from a hexylagarose column then analyzed by Western blot; it reacted with bovine and human cysticercosis infection sera and with specific monoclonal antibodies but not with control sera or fascioliasis infection sera. When the eluate was used as coating antigen in a plate ELISA assay no false positive reactions were seen in sera from cattle infected with Fasciola hepatica; false positive reactions were observed for the unfractionated ThFAS antigen preparation.

Animals↗

Molecular vaccines against parasites.

Prophylactic vaccines can be expected to be one of the major practical outputs of parasitology research. Various groups within Australia have pursued the vaccine objective for several years, with particular emphasis on blood-stage falciparum malaria in man, intestinal helminths of sheep and cattle, cutaneous myiasis (blowfly strike) in sheep, cysticercosis in sheep and cattle, bovine babesiosis, and cattle ticks. Other vaccine programmes are concerned with giardiasis, filariasis, toxoplasmosis, fascioliasis, coccidiosis in poultry, cutaneous leishmaniasis and schistosomiasis japonica. For many years, the only available vaccine against a parasite in Australia has been the attenuated Babesia bovis vaccine produced by the Tick Fever Research Centre of the Queensland Department of Primary Industries. Strategies for achieving molecular vaccines are generally similar within the various research groups. They involve analysis of the immunology and immunochemistry of a model or in-vitro system; development of functional monoclonal antibodies; analysis of antibody specificities in clinically and/or functionally defined polyclonal sera; screening of cDNA or genomic expression libraries; peptide synthesis; identification of an appropriate vaccination schedule involving adjuvants or new recombinant DNA-based antigen delivery systems. Outlined below are five of the major vaccine programmes.

Journal Article↗

Geographic models and control of cattle liver flukes in the Southern USA.

John Malone and Stanley Zukowski describe climate forecast and soil hydrology-based geographic information system models of snail habitat extent that provide an environmental context for site-specific mathematical models of fascioliasis. These have potential for the development of broad-scale control models that include a cost-benefit analysis component.

Journal Article↗

Recent applications of the Dot-ELISA in immunoparasitology.

The dot enzyme-linked immunosorbent assay (Dot-ELISA) is a highly versatile solid-phase immunoassay for antibody or antigen detection. The assay uses minute amounts of reagent dotted onto solid surfaces such as nitrocellulose and other paper membranes which avidly bind proteins. After incubation with antigen-specific antibody and enzyme-conjugated anti-antibody, the addition of a precipitable, chromogenic substrate causes the formation of a colored dot on the solid phase which is visually read. The Dot-ELISA has been used extensively in the detection of human and veterinary protozoan and metazoan parasitic diseases, including amebiasis, babesiosis, fascioliasis, cutaneous and visceral leishmaniasis, cysticercosis, echinococcosis, malaria, schistosomiasis, toxocariasis, toxoplasmosis, trichinosis, trypanosomiasis and even ixodid tick infestation. The technique is rapid, easy to perform and interpret, reagent conservative, cost effective and field portable. In addition, the Dot-ELISA may be configured to detect antibodies or parasite antigen in either microtiter plates for large-batch testing or with dipsticks for small numbers of determinations. A slight modification of the Dot-ELISA procedure allows the determination of infection rates of vectors such as ticks and sandflies with parasites.

Animals↗

Waterborne zoonotic helminthiases.

This review deals with waterborne zoonotic helminths, many of which are opportunistic parasites spreading directly from animals to man or man to animals through water that is either ingested or that contains forms capable of skin penetration. Disease severity ranges from being rapidly fatal to low-grade chronic infections that may be asymptomatic for many years. The most significant zoonotic waterborne helminthic diseases are either snail-mediated, copepod-mediated or transmitted by faecal-contaminated water. Snail-mediated helminthiases described here are caused by digenetic trematodes that undergo complex life cycles involving various species of aquatic snails. These diseases include schistosomiasis, cercarial dermatitis, fascioliasis and fasciolopsiasis. The primary copepod-mediated helminthiases are sparganosis, gnathostomiasis and dracunculiasis, and the major faecal-contaminated water helminthiases are cysticercosis, hydatid disease and larva migrans. Generally, only parasites whose infective stages can be transmitted directly by water are discussed in this article. Although many do not require a water environment in which to complete their life cycle, their infective stages can certainly be distributed and acquired directly through water. Transmission via the external environment is necessary for many helminth parasites, with water and faecal contamination being important considerations. Human behaviour, particularly poor hygiene, is a major factor in the re-emergence, and spread of parasitic infections. Also important in assessing the risk of infection by water transmission are human habits and population density, the prevalence of infection in them and in alternate animal hosts, methods of treating sewage and drinking water, and climate. Disease prevention methods, including disease surveillance, education and improved drinking water treatment are described.

Animals↗

The impact on database searching arising from inconsistency in the nomenclature of parasitic diseases.

The existing usage of disease names formed from the name of the parasite taxon is characterised by marked heterogeneity. This is largely due to the fact that, for coining disease names, four different suffixes, '-osis', '-iosis', '-asis' and '-iasis', are being used inconsistently. The result is that alternative terms are in use for naming the same disease, e.g. trypanosomosis and trypanosomiasis, fasciolosis and fascioliasis, ascariosis and ascariasis. Inspite of the SNOAPAD/SNOPAD guideline (1988) which proposed the principles of a uniform and standard disease nomenclature, the actual usage depends largely on tradition, educational imprinting and personal preferences, showing great variation. By using alternative disease names as search terms the author investigates in four databases the impact of nomenclatural heterogeneity on information storage and retrieval. It is evident that the existence of alternative disease names in parasitology markedly interferes with the efficacy of online data retrieval. The value of a disease name as a search term was shown to be greatly different in various databases. Until we have to coexist with an inconsistent disease terminology we need to adopt specially structured database-search techniques to ensure a proper level of precision in searching. Such possible techniques are considered.

Animals↗

Identification and characterization of a cathepsin L-like cysteine protease from Taenia solium metacestode.

Taenia solium metacestode, a larval pork tapeworm, is a causative agent of neurocysticercosis, one of the most common parasitic diseases in the human central nervous system. In this study, we identified a cDNA encoding for a cathepsin L-like cysteine protease from the T. solium metacestode (TsCL-1) and characterized the biochemical properties of the recombinant enzyme. The cloned cDNA of 1216 bp encoded 339 amino acids with an approximate molecular weight of 37.6 kDa which containing a typical signal peptide sequence (17 amino acids), a pro-domain (106 amino acids), and a mature domain (216 amino acids). Sequence alignments of TsCL-1 showed low sequence similarity of 27.3-44.6 to cathepsin L-like cysteine proteases from other helminth parasites, but the similarity was increased to 35.9-55.0 when compared to mature domains. The bacterially expressed recombinant protein (rTsCL-1) did not show enzyme activity; however, the rTsCL-1 expressed in Pichia pastoris showed typical biochemical characteristics of cysteine proteases. It degraded human immunoglobulin G (IgG) and bovine serum albumin (BSA), but not collagen. Western blot analysis of the rTsCL-1 showed antigenicity against the sera from patients with cysticercosis, sparganosis or fascioliasis, but weak or no antigenicity against the sera from patients with paragonimiasis or clonorchiasis.

Amino Acid Sequence↗

The School Health Programme in Behera: an integrated helminth control programme at Governorate level in Egypt.

School Health Programmes offer the opportunity to deliver public health interventions to a great number of beneficiaries at a relatively low cost and are seen with growing interest by policy makers in developing countries and the donors' communities. In Egypt a pilot School Health Programme has been implemented for the past 6 years in Behera, the largest and most populous Governorate of the Nile Delta. The Programme integrated additional activities for the control of soil-transmitted helminthiasis, human fascioliasis and anaemia in the National Schistosomiasis Control Programme (NSCP), implemented in Egypt since 1988 by the Ministry of Health and Population (MoHP). To facilitate planning and direct actions, a strong monitoring system was also developed, that generated useful information for the schistosomiasis control programme. The practical steps adopted to develop the programme are presented and discussed. Results from three rounds of monitoring (2000, 2001, 2002) are analysed and compared with baseline data conducted in 1996, together with the cost of each activity. On the basis of the experience gained by the Behera School Health Programme a number of operational recommendations are formulated.

Adolescent↗

Emerging parasite zoonoses associated with water and food.

The environmental route of transmission is important for many protozoan and helminth parasites, with water, soil and food being particularly significant. Both the potential for producing large numbers of transmissive stages and their environmental robustness, being able to survive in moist microclimates for prolonged periods of time, pose a persistent threat to public and veterinary health. The increased demands on natural resources increase the likelihood of encountering environments and produce contaminated with parasites. For waterborne diseases, the protozoa, Cryptosporidium, Giardia and Toxoplasma, are the most significant causes, yet, with the exception of Toxoplasma, the contribution of zoonotic transmission remains unclear due to the absence of 'standardised' methods. The microsporidia have been documented in one waterborne outbreak, but the role of animals as the cause of contamination was not elucidated. In foods, surface contamination is associated with the faecal-oral pathogens, and some data are available to indicate that animal wastes remain an important source of contamination (e.g. cattle faeces and apple cider outbreaks), however, further work should focus on examining the source of contamination on fruit and vegetables. Increasing recognition of the burden of human fascioliasis has occurred; it is now recognised as an emerging zoonosis by the WHO. Toxoplasma, Trichinella and Taenia spp. remain important meatborne parasites, however, others, including Pleistophora-like microsporidians may be acquired from raw or lightly cooked fish or crustaceans. With increased international travel, the public health importance of the foodborne trematodiases must also be realised. Global sourcing of food, coupled with changing consumer vogues, including the consumption of raw vegetables and undercooking to retain the natural taste and preserve heat-labile nutrients, can increase the risk of foodborne transmission. A greater awareness of parasite contamination of our environment and its impact on health has precipitated the development of better detection methods. Robust, efficient detection, viability and typing methods are required to assess risks and to further epidemiological understanding.

Animals↗

Demonstration of a specific Echinococcus multilocularis antigen in the supernatant of in vitro maintained protoscolices.

Serodiagnosis of echinococcosis is still often met with difficulties resulting from unspecific reactions due to crude antigens which contain numerous host-derived proteins. In order to eliminate host protein contamination we produced Echinococcus multilocularis antigen by methods of in vitro technique: Evaginated protoscolices of Echinococcus multilocularis isolated from experimentally infected Mongolian gerbils (Meriones unguiculatus) were maintained in RPMI 1640 medium. Although no serum proteins were added, protoscolices could be kept alive for more than 2 weeks. The supernatants harvested from protoscolices cultures were tested for their immunoreactivity against sera of patients with confirmed alveolar or cystic echinococcosis, cysticercosis, schistosomiasis or fascioliasis by means of SDS-PAGE and immunoblotting. A polypeptide band at about 62,000 mol. mass was identified which proved to be specifically immunoreactive with sera from patients with alveolar echinococcosis, whereas sera from patients with cystic echinococcosis or with other helminthic infections as well as sera from healthy blood-donors did not react with this polypeptide band. Thus, the use of supernatant antigen in immunoblotting technique allows a serological differentiation between infections with Echinococcus multilocularis and those with Echinococcus granulosus and provides an accurate diagnosis of alveolar echinococcosis.

Animals↗

Focal inflammatory diseases of the liver.

Inflammatory lesions constitute an important subgroup of focal liver lesions. They may mimic primary or metastatic neoplastic lesions and their differentiation from neoplasia is clinically very important since management of the patient significantly changes. Radiologists should have an important role in both the diagnosis and therapy of these lesions by performing percutaneous aspirations and drainages. In this review we discussed the radiological findings of pyogenic abscesses, amebic abscesses, candidiasis, tuberculosis, hydatic cysts, fascioliasis, ascariasis, schistosomiasis, and sarcoidosis with a special emphasis on US, CT and MR characteristics.

Diagnostic Imaging↗

Endoscopic diagnosis and management of tropical parasitic infestations.

Biliary parasitosis is one of the common causes of biliary obstruction in developing countries and can often be confused with stone disease. With increased worldwide travel and immigration, these conditions are not limited to the developing countries alone. Ascariasis, hydatid liver disease, clonorchiasis, and fascioliasis are the commonly encountered parasitic infestations of the biliary tract usually presenting with biliary colic or cholangitis. Endoscopy has an important role in the diagnosis and emergent management of these conditions and in elective endoscopic therapy of associated complications. Endoscopic sphincterotomy and bile ductal clearance, along with pharmacotherapy, are the mainstays of treatment.

Biliary Tract Diseases↗