Expression and characterization of glutathione peroxidase of the liver fluke, Fasciola gigantica.

Glutathione peroxidase (GPx) is a key member of the family of antioxidant enzymes in trematode parasites including Fasciola spp. Because of its abundance and central role as an anti-oxidant that helps to protect parasites from damage by free radicals released from the host immune cells, it has both diagnostic as well as vaccine potential against fasciolosis. In this study, we have cloned, characterized, and detected the expression of the GPx protein in Fasciola gigantica (Fg). FgGPx (582 bp) was cloned by polymerase chain reaction (PCR) from complementary DNA (cDNA) from an adult fluke. Its putative peptide has no signal sequence and is composed of 168 amino acids, with a molecular weight of 19.1 kDa, and conserved sequences at NVACKUG, FPCNQFGGQ, and WNF. Phylogenetic analysis showed that GPx is present from protozoa to mammals and FgGPx was closely related to Fasciola hepatica GPx. A recombinant FgGPx (rFgGPx) was expressed in Escherichia coli BL21 (DE3) and used for immunizing mice to obtain polyclonal antibodies (anti-rFgGPx) for immunoblotting and immunolocalization. In immunoblotting analysis, the FgGPx was expressed in all stages of F. gigantica (eggs, metacercariae, newly excysted juveniles (NEJ), 4-week-old juveniles, and adults). This mouse anti-rFgGPx reacted with the native FgGPx at a molecular weight of 19.1 kDa in adult whole body (WB) and tegumental antigens (TA) as detected by immunoblotting. The FgGPx protein was expressed at a high level in the tegument, vitelline glands, and eggs of the parasite. Anti-rFgGPx exhibited no cross-reactivity with the other parasite antigens, including Eurytrema pancreaticum, Cotylophoron cotylophorum, Fischoederius cobboldi, Gastrothylax crumenifer, Paramphistomum cervi, and Setaria labiato papillosa. The possibility of using rFgGPx for immunodiagnosis and/or as a vaccine for fasciolosis in animals of economic importance will be explored in the future.

Liver Fluke-Associated Biliary Tract Cancer.

Cholangiocarcinoma (CCA) is an aggressive cancer arising from epithelial cells of the bile duct. Most patients with CCA have an unresectable tumor at the time of diagnosis. In Western countries, the risk of CCA increases in patients with primary sclerosing cholangitis, whereas liver fluke infection appears to be the major risk factor for CCA in Asian countries. A diagnosis of liver fluke infection often relies on stool samples, including microscopic examination, polymerase chain reaction-based assays, and fluke antigen detection. Tests of serum, saliva and urine samples are also potentially diagnostic. The presence of liver fluke along with exogenous carcinogens magnifies the risk of CCA in people living in endemic areas. The "liver fluke-cholangiocarcinoma" carcinogenesis pathways consist of mechanical damage to the bile duct epithelium, immunopathologic and cellular reactions to the liver fluke's antigens and excretory/secretory products, liver fluke-induced changes in the biliary tract microbiome and the effects of repeated treatment for liver fluke. A vaccine and novel biomarkers are needed for the primary and secondary prevention of CCA in endemic areas. Importantly, climate change exerts an effect on vector-borne parasitic diseases, and awareness of liver fluke should be enhanced in potentially migrated habitat areas.

Hepatic fascioliasis presenting with bile duct obstruction: a case report.

Fascioliasis is a zoonotic infection caused by a liver trematode: fasciola hepatica; which commonly affects cattle and sheep, humans are accidental hosts. Several cases have been reported in the literature worldwide with a large geographical distribution. We present a case of bile duct obstruction due to a hepatic fascioliasis, successfully treated with both a combined surgical and medical approaches. A high index of suspicion should be kept in mind for all cases of obstructive jaundice, especially in areas in which human fascioliasis infection is repeatedly reported.

Human and livestock trematode infections in a mobile pastoralist setting at Lake Chad: added value of a One Health approach beyond zoonotic diseases research.

Background: At Lake Chad in Central Africa, mobile pastoralists face economic losses due to livestock trematodiases. Fasciola gigantica and Schistosoma bovis-trematodes that affect livestock-share transmission ecology traits with Schistosoma haematobium and S. mansoni that cause human schistosomiasis. This project aimed at assessing treatment strategies and elucidating the predictive potential of human and livestock trematode infections. Methods: Schistosomiasis and fascioliasis were investigated concurrently in humans and cattle by repeated cross-sectional surveys. Urine and stool samples from humans and faecal samples from cattle were examined for trematode eggs. Treatment strategies were assessed by means of focus group discussions and in-depth interviews. Results: Mobile pastoralists of four ethnic groups participated. Prevalence of human schistosomiasis and livestock trematodiases showed considerable heterogeneity from one ethnic group to another, but correlated within ethnic groups. Effective trematocidal drugs were not available in the study area. Conclusions: Mutual predictive potential of human schistosomiasis and livestock fascioliasis relates to distinct livestock husbandry practices. Introducing efficacious strategic treatment against human schistosomiasis and livestock fascioliasis might improve human and animal health and well-being. Our research provides evidence for the benefits of a One Health approach targeting diseases that share specific ecological traits.

Caracterización clínica y epidemiológica de la infección por fasciola hepática entre los años 2003-2010 en el Hospital Nacional Cayetano Heredia, Lima, Perú / Demographic and clinical aspects of hepatic fascioliasis between 2013-2010 in National Hospital Cayetano Heredia, Lima, Peru

Describir la distribución demográfica y clínica de la fasciolosis hepática, así como las complicaciones y asociaciones entre diversos factores y la enfermedad en un hospital de referencia. Material y métodos: Estudio descriptivo y retrospectivo de tipo serie de casos, se incluyeron todos los pacientes con diagnóstico de fasciolosis hepática por primera vez entre 2003 a 2010 en el Hospital Nacional Cayetano Heredia, se analizaron variables demográficas y clínicas incluyendo las complicaciones y el tratamiento. Resultado: Se encontraron 68 historias clínicas aptas para el estudio, el promedio de edad fue 36 años, Ancash fue el departamento de nacimiento y procedencia más frecuente. La mayoría se diagnosticó en fase crónica, el síntoma más frecuente fue dolor abdominal y se presentó complicaciones en 6 pacientes: 3 abscesos hepáticos, 1 hematoma subcapsular, 1 colangitis y 1 colangitis más colecistitis. Se encontraron las siguientes asociaciones: edad menor de 15 años con la fase crónica e hiporexia, ser estudiante con un coprológico positivo; y entre el nacimiento de una zona endémica con la ausencia de desarrollar complicaciones. Conclusiones: La fasciolosis hepática tiene una presentación clínica poco específica, por lo que los antecedentes epidemiológicos o alimentarios, y especialmente la eosinofilia deben orientar el diagnóstico...

To describe the demographic and clinical aspects of hepatic fascioliasis as well as the complications and associations between various factors and the disease in a reference hospital. Methods: This is a descriptive and retrospective case series study; we included all patients who had a recent diagnosis of hepatic fascioliasis from 2003 to 2010 in the Hospital Nacional Cayetano Heredia. Demographic and clinical variables were analyzed including complications and treatment received. Results: 68 medical records were found eligible for the study. The mean age was 36 years. Ancash department was the most frequent place of origin and residence. Most of them were diagnosed in the chronic phase, the most common symptom was abdominal pain and eight patients had complications: 3 hepatic abscess, 1 subcapsular hematoma, 1 cholangitis and 1 cholangitis plus cholecystitis. There were the following associations: Age under 15 years with chronic phase and hyporexia, being a student with a positive stool analysis; and between being born in an endemic area with the absence of complications. Conclusions: Hepatic fascioliasis has unspecific clinical presentation so the epidemiological or dietary history and specially eosinophilia should guide the diagnosis...

Biliary colic with dilated common bile duct: simple "sheepish" problem?

A 40-year-old man, resident of a rural area of Rajasthan state of India presented with recurrent biliary colic and fever since last 2 years. Examination was normal and investigations revealed a dilated common bile duct with elevated alkaline phosphatase. Magnetic resonance cholangiopancreatography revealed a dilated biliary system without any evident cause. Patient was subjected to endoscopic retrograde cholangioancreatography and it demonstrated biliary dilatation with a linear filling defect in lower common bile duct, further sphinicterotomy was done and a live Fasciola hepatica was extracted using a biliary basket.

Direct and indirect affection of the central nervous system by Fasciola infection.

Fascioliasis is a worldwide, zoonotic disease caused by the liver trematodes Fasciola hepatica and Fasciola gigantica. Neurological fascioliasis has been widely reported in all continents, affecting both sexes and all ages. Two types of records related to two physiopathogenic mechanisms may be distinguished: cases in which the neurological symptoms are due to direct effects of a migrating juvenile present in the brain or neighboring organ and with cerebral lesions suggesting migration through the brain; and cases with neurological symptoms due to indirect immuno-allergic and toxic effects at distance from flukes in the liver. Neurological manifestations include minor symptoms, mainly cephalalgias, and major symptoms which are nonspecific, extremely diverse, varying from one patient to another and even within the same patient, and comprising meningeal manifestations and impressive neurological manifestations. The puzzling neurological polymorphism leads to confusion with cerebral tumors, multiple sclerosis, lesions of the brainstem, or cerebro-meningeal hemorrhages. Only blood eosinophilia and information on infection source guide toward correct diagnosis by appropriate coprological and/or serological techniques. Although neurological patients usually recover after fasciolicide treatment or surgical worm extraction, sequelae, which are sometimes important, remain in several patients. The need to include possible neurological complications within the general frame of fascioliasis becomes evident.

Obstrucción biliar por fascioliasis hepática / Biliary obstruction due to hepatic fascioliasis

Hepatic fascioliasis is produced by a platyhelminth, trematode: the hepatic Fasciola. In recent years, its incidence has increased in all countries, including Latin America. The diagnosis is based on clinical and epidemiological background information, hemogramm with eosinophil count; Ig G ELISA; and copro-parasitological and duodenal aspirate in search of parasite eggs. Images are useful, endoscopic ultrasound, Computed Tomography (CT), abdominal Magnetic Resonance Imaging (MRI), especially in case of complications. The endoscopic retrograde cholangiopancreatography (ERCP) is useful both for diagnosis and treatment (extraction of adult Fasciola), as it was in hte present clinical case. This parasite can also be found incidentally in the course of a surgery planned for other reasons. In the present clinical case we present a 41-year-old patient, farmer, with apparent ingestion of water and contaminated aquatic vegetables. Initially he presented only fever and hypereosinophilia; with fascioliasis diagnosis. The pathology recurred after 18 month with biliary obstruction. The clinical and epidemiological background information was a fundamental tool for diagnosis, together with abdominal CT. Initially it evolved with bad response to several antiparasitic treatments and then it progressed positively. After that, the pathology recurred with complications. ERCP was performed to extract adult Fasciola. Currently in symptomatic remission after endoscopic intervention and drug treatment.

La fascioliasis hepática es producida por un platelminto, tremátodo: la Fasciola hepática. En los últimos años ha incrementado su incidencia en todos los países, incluyendo América Latina. El diagnóstico se basa en los antecedentes clínicos, epidemiológicos, hemograma con recuento de eosinófilos; IgG Elisa; copro-parasitológicos y aspirado duodenal en búsqueda de huevo de parásitos. Las imágenes son de utilidad, endosonografía, tomografía computada (TC), resonancia magnética (RM) abdominal, especialmente en caso de complicaciones. La colangio-pancreatografía retrógrada endoscópica (CPRE) es útil tanto como para el diagnóstico como para el tratamiento (extracción de fasciolas adultas), como se realizó en el presente caso clínico. Este parásito también se puede encontrar incidentalmente en el transcurso de una cirugía por otros motivos. Presentamos el caso de un paciente de 41 años, originario de una zona rural, agricultor, con ingesta aparente de agua y vegetales acuáticos contaminados. Cursó inicialmente sólo con fiebre e hipereosinofilia; con diagnóstico de fascioliasis. Al cabo de 18 meses su patología recurrió con cuadro clínico de obstrucción biliar. El antecedente clínico y epidemiológico fue una herramienta fundamental para el diagnóstico, además, de la TC abdominal. Evolucionó inicialmente con mala respuesta a varios tratamientos antiparasitarios, y luego mejoró. Posteriormente, su patología recurrió con complicaciones. Se realizó CPRE con extracción de Fasciolas adultas. Actualmente, se encuentra en remisión sintomática mantenida, después de la intervención endoscópica y el tratamiento medicamentoso.

Fasciolosis aguda: presentación de un caso de Cuba / Acute fasciolosis: a Cuban case report

Fasciolosis es una enfermedad parasitaria de humanos y diferentes animales. Sus agentes causales son Fasciola hepatica y Fasciola gigantica; sin embargo, en Cuba, solo la primera de esas especies ha sido notificada. Como otras enfermedades incorporadas en el grupo de las olvidadas o desatendidas, fasciolosis, es más frecuente en personas que viven en áreas rurales donde coexisten animales infectados, hospederos intermediarios y hospederos susceptibles. Se exponen las características de un caso que pudiera contribuir en el proceso de educación continuada de los profesionales de la salud en nuestra área geográfica(AU)

Fasciolosis is a worldwide parasitic disease in human and domestic animals. The causative agents of fasciolosis are Fasciola hepatica and Fasciola gigantica but, in Cuba, only F. hepatica has been notified. As others neglected diseases, F. hepatica is more common in people living in rural areas where infected animals, intermediate hosts and susceptible hosts coexist. We describe the clinical features of a case that could contribute to the process of continuing medical education of health professionals in our geographic area(AU)

Biliary fascioliasis--an uncommon cause of recurrent biliary colics: report of a case and brief review.

Biliary parasitosis is one of the important causes of biliary obstruction in endemic areas, however due to migration and travel the disease is known to occur in non endemic zones as well. The spectrum of biliary fascioliasis ranges from recurrent biliary colics to acute cholangitis. The long term complications are gall stones, sclerosing cholangitis and biliary cirrhosis. We describe fascioliasis as a cause of recurrent biliary colics in a young male necessitating multiple hospitalizations over a period of four years. Investigative profile had been non-contributory every time he was hospitalized for his abdominal pain prior to the current presentation. He never had cholangitis due to the worm in the common bile duct. It was only at endoscopic retrograde cholangiopancreatography (ERCP) biliary fascioliasis was discovered to be the cause of his recurrent biliary colics. After removal of the live Fasciola hepatica from the common bile duct he became symptom free and is attending our clinic for last 11 months now. Clinical spectrum of biliary fascioliasis is discussed in this report.

Fungus-benzimidazole interactions: a prerequisite to deploying egg-parasitic fungi Paecilomyces lilacinus and Verticillium chlamydosporium as biocontrol agents against fascioliasis and amphistomiasis in ruminant livestock.

In vitro trials investigating the effects of albendazole and triclabendazole anthelmintics on the growth profiles of the egg-parasitic fungi Paecilomyces lilacinus and Verticillium chlamydosporium were undertaken. In addition, in vivo trials were conducted in goats fed on millet grain cultures of each fungus and administered albendazole and triclabendazole anthelmintics. In vitro growth revealed V. chlamydosporium to be more sensitive to albendazole compared to P. lilacinus. In contrast, triclabendazole had the least inhibitory effect on in vitro growth of both P. lilacinus and V. chlamydosporium. Similar to albendazole, growth of P. lilacinus was more vigorous at 0.5 ppm concentration of triclabendazole. Efforts to re-isolate these egg-parasitic fungi from faeces of goats fed on fungal millet grain cultures before and following single intraruminal administration of albendazole and triclabendazole showed that P. lilacinus was not able to be re-isolated from the faeces at any sampling period. In contrast, V. chlamydosporium was able to be re-isolated from the faeces at all of the sampling periods except for the samples taken at 8-18 h and 18-24 h after administration of albendazole and triclabendazole, respectively. Lack of fungal activity at these times coincided with peak plasma availability of anthelmintics and suggests faecal levels of drugs were also high at these times and impacted negatively on fungal viability.

Hematoma subcapsular por Fasciolasis / Liver subcapsular hematoma caused by Fasciola hepatica: report of one case

We report a 60 years old female admitted for pain in the right upper quadrant of the abdomen, vomiting and fever. Initial laboratory showed leukocytosis with 56 percent of eosinophils. An abdominal CAT scan showed a subcapsular fluid collection in the liver. The patient was operated and the presence of a subcapsular hematoma was confirmed. Blood serological tests were positive for Fasciola hepatica. The patient was treated with tricabendazole and after one year of follow up, is in good conditions.

Presentamos el caso de una paciente de sexo femenino de 60 años de edad, ingresada en nuestro centro hospitalario por cuadro agudo de dolor abdominal en hipocondrio derecho asociado a vómitos y fiebre. Destaca la presencia de leucocitosis con eosinofilia de 56 por ciento. La tomografía computarizada abdominal muestra una colección subcapsular hepática. Durante su evolución presenta abdomen agudo, realizándose una laparoscopía exploratoria que confirma un hematoma subcapsular con presencia de hemopentoneo de escasa cuantía. La serología confirma infección por fasciola hepática. Se realizó su tratamiento con triclabendazol en monodosis. Además, realizamos una revisión de la literatura nacional e internacional sobre el tema, sus manifestaciones clínicas, métodos diagnósticos y tratamiento.

The silencing of cysteine proteases in Fasciola hepatica newly excysted juveniles using RNA interference reduces gut penetration.

Probing protein function in parasitic flatworms is hampered by the difficulties associated with the development of transgenic approaches. Although RNA interference (RNAi) in schistosomes shows much promise, it has not been reported in other trematodes. Here, we show the successful silencing of the cysteine proteases cathepsin B and L in the infective stage of Fasciola hepatica newly excysted juveniles (NEJs). Silencing resulted in marked reductions in target transcript levels and significant diminution in the encoded proteins in the gut. RNAi of either enzyme in NEJs induced transient, abnormal locomotory phenotypes, and significantly reduced penetration of the rat intestinal wall.

Hematoma subcapsular hepático por fasciolasis / Hepatic subcapsular hematoma caused by fascioliasis

Hepatic fasciolasis is a worldwide spread zoonoses mainly affecting cattle-raising countries. It is caused by the trematode Fasciola hepßtica and it is characterized by abdominal pain, fever, nausea and vomitus, weight loss, diahrrea, paleness, general malaise, and hypereosinophilia. Immunological diagnosis as well as stool eggs count may be performed. Hepatic subcapsular and intraparenchymatous hematoma is an infrequent complication of human fascioliasis. Nevertheless, for establishing a proper diagnosis and treatment, any suspicion of its presence must be carefully discarded through, clinical epidemiology, laboratory and imaging exams and procedures. The aim of this study is to expand knowledge on this unfrequently dealt pathology in medical literature by presenting four case reports related to patients undergoing a two-year treatment. All of them had been referred from Departamento de Cajamarca, Peru.

La fasciolasis hepática es una zoonosis mundialmente difundida, sobre todo en los países productores de ganado; causada por la fasciola hepática. Se manifiesta por dolor abdominal, fiebre, nauseas y vómitos, baja de peso, diarrea, palidez, malestar general e hipereosinofilia. El diagnóstico es inmunológico y también puede hacerse por recuento de huevos en heces. El hematoma subcapsular e intraparenquimatoso hepático es una complicación rara de la fasciolasis humana pero se debe tener un alto índice de sospecha uniendo epidemiología, clínica, laboratorio e imaginologia para un adecuado diagnóstico y tratamiento. El propósito de reportar estos casos es dar a conocer una patología poco frecuente en la literatura, con una casuística de cuatro pacientes tratados en el lapso de dos años, todos referidos del Departamento de Cajamarca.