Acid-Fast Staining for Hooklets of Echinococcus

Recognition of well-developed hydatid disease in tissue sections or in aspirated cyst fluid usually presents no difficulties. The distinctively laminated outer capsule, the active inner germinative layer and the hydatid fluid containing characteristic brood capsules, scolices and free hooklets, are outstandingly reliable diagnostic criteria. The identification of the organism assumes more difficulty in old, arrested infections when the outer layer, and even the surrounding cellular inflammatory response, has been replaced by a wall of nonspecific, collagenous tissue and the cyst contents contain only calcified remnants of scolices. In the course of time, even the pathognomonic hooklets may no longer be recognizable.     

Hydatid cyst in soft tissues mimicking malignant tumors. Diagnosis by fine needle aspiration cytology

OBJECTIVE: To evaluate the role of fine needle aspiration cytology in the diagnosis of soft tissue hydatid cysts. STUDY DESIGN: Five cases of soft tissue hydatid cyst were diagnosed primarily by fine needle aspiration cytology. RESULTS: In all cases, large fragments of acellular material, finely lamellated, were found. There were no complications related to fine needle aspiration, and histologic studies confirmed the diagnosis of hydatid cyst. CONCLUSION: When acellular, laminated fragments suggestive of a laminated layer are identified on smears, hydatid cyst should be considered in the differential diagnosis, even in atypical locations and in the absence of hooklets, protoscolices or both.     

Cytologic findings in aspirated hydatid fluid

A case of hydatid disease of the liver diagnosed by accidental needle aspiration of a cyst is reported. The patient presented with multiple symptoms, whose cause could not be readily determined. Since an amoebic abscess of the tender and enlarged liver was one possibility, that organ was aspirated with an 18-gauge needle, producing about 200 mL of slightly turbid fluid; the patient went into anaphylactic shock following the procedure, but was revived. Smears of the fluid showed brood capsules, free scolices and scattered hooklets pathognomonic of Echinococcus. The cytologic diagnosis of echinococcosis was confirmed by histopathologic examination of the surgically removed cysts.     

Intracranial hydatidosis. Report of a case diagnosed on cerebrospinal fluid cytology

BACKGROUND: Hydatidosis occurs due to infestation with the larval stage of the Echinococcus species. Humans are incidental hosts. Symptoms and signs in humans are a slowly growing mass lesion, especially in the central nervous system. Diagnosis depends on the appropriate presentation and history with corroborative radiology and serology. Microscopic detection of characteristic parasitic scolices and hooklets is confirmatory. CASE: A 10-year-old girl presented with complaints of headaches and convulsions for the preceding several months. Computed tomography findings showed asymmetric dilatation of the right lateral ventricle by an intraventricular, focally calcified mass. The radiologic suspicion was an intraventricular tuberculoma, hydatid cyst or choroid plexus papilloma. Cytologic examination of cerebrospinal fluid (CSF) smears revealed characteristic echinococcal components. CONCLUSION: Intracranial hydatid cyst is a fairly uncommon manifestation of an echinococcal infestation and shares clinicoradiologic features with several intracranial, space-occupying lesions. Cytologic examination of CSF samples may be employed to detect confirmatory evidence of a clinical/radiologic suspicion.     

Pelvic Hydatidosis Mimicking a Malignant Multicystic Ovarian Tumor

Echinococcosis is a multisystem disease and has propensity to involve any organ, an unusual anatomical site, and can mimic any disease process. Primary peritoneal echinococcosis is known to occur secondary to hepatic involvement but occasional cases of primary peritoneal hydatid disease including pelvic involvement have also been reported. We report here 1 such case of primary pelvic hydatidosis mimicking a malignant multicystic ovarian tumor where there was no evidence of involvement of the liver or spleen. Our patient, a 27-year-old female, was detected to have a large right cystic adnexal mass on per vaginal examination which was confirmed by ultrasonography. Her biochemical parameters were normal and CA-125 levels, though mildly raised, were below the cut off point. She underwent surgery and on exploratory laparotomy, another cystic mass was found attached to the mesentery of the small gut. The resected cysts were processed histopathologically. On cut sections both large cysts revealed numerous daughter cysts. Microscopic examination of fluid from the cysts revealed free scolices with hooklets and the cyst wall had a typical laminated membrane with inner germinal layer containing degenerated protoplasmic mass. The diagnosis of pelvic hydatid disease was confirmed and patient was managed accordingly. Hydatid disease must be considered while making the differential diagnosis of pelvic cystic masses, especially in endemic areas.     

An Imported Case of Echinococcosis of the Liver in a Korean Who Traveled to Western and Central Europe

Echinococcus granulosus, an intestinal tapeworm of dogs and other canids, infects humans in its larval stage and causes human echinococcosis or hydatid disease. In the Republic of Korea, 31 parasite-proven human echinococcosis cases have been reported, most of which were imported from the Middle East. We recently examined a 61-year-old Korean man who had a large cystic mass in his liver. ELISA was negative for tissue parasitic infections, including echinococcosis, cysticercosis, paragonimiasis, and sparganosis. The patient underwent surgery to remove the cyst, and the resected cyst was processed histopathologically for microscopic examinations. In sectioned cyst tissue, necrotizing protoscolices with disintegrated hooklets of E. granulosus were found. In some areas, only freed, fragmented hooklets were detected. The patient had traveled to western and central Europe in 1996, and had no other history of overseas travel. We report our patient as a hepatic echinococcosis case which was probably imported from Europe.     

Purification and partial immunochemical characterization of a low molecular mass, diagnostic Echinococcus granulosus immunogen for sheep hydatidosis

Abstract A hydatid specific antigen of 8 kDa molecular mass was affinity-purified from crude hydatid cyst fluid. Some of the epitopes recognised by antibodies in the sera from sheep with hydatidosis were periodate-sensitive. The purified 8 kDa antigen was observed to be a thermo-stable glycoprotein in its immunochemical characteristics. By immunofluorescence on acetone-fixed protoscolices anti-8 kDa monospecific IgG antibodies indicated the existence of the 8 kDa molecule on the hooklets of protoscolices. The purified antigen was used in an enzyme-linked immunosorbent assay for the detection of specific antibodies in sera from sheep hydatidosis. Eighteen (90%) of 20 sera from sheep hydatidosis had antibodies to purified 8 kDa antigen while none of the sera from other parasitic infections or uninfected animals had any detectable levels of antibodies to 8 kDa antigen. Thus, the data on localization and recognition of hydatid specific 8 kDa molecule suggested that this may be one of the major molecules for specific immunodiagnosis and for modulating the hydatid disease process in infected hosts.     

Echinococcosis (Hydatid Disease): Review and Report of a Case of Secondary Echinococcosis

Echinococcosis is a disease caused by the larval form of Echinococcus granulosus. The adult worm lives parasitically in the intestine of the definitive host, the dog, wolf and other wild carnivores. The ova, which are passed in the feces, are ingested by grazing animals. The intestinal juices free the ovum from its cuticle and, having entered a capillary, it is carried by the blood stream to the liver or lung, where it develops into a hydatid cyst. Man becomes infected by ingesting contaminated vegetables or drinking contaminated water.In a mature hydatid cyst there may be thousands of scolices and each one of these, if released from the hydatid cyst, may form a new cyst if it is planted in a suitable environment. Hydatid cysts are most commonly found in the liver and the lung, but may occur in other organs.A hepatic cyst may rupture into the peritoneal cavity and produce a hydatidoperitoneum. A pulmonary cyst frequently ruptures into a bronchus and is thus evacuated.The disease is of world-wide distribution. In Canada most of the cases of this disease have occurred among Indians and Eskimos. It is now known that a sylvatic cycle has been established in North America and that it serves as a source of infection of the dog and man.The patient reported herein had spontaneous rupture of a hepatic hydatid cyst, with subsequent development of secondary peritoneal echinococcosis. Repeated laparotomies had to be performed to relieve pressure on or obstruction of various organs. The patient made an uneventful recovery and is again usefully employed. No effective medical treatment for this disease is available.     

Proteomic analysis of the Echinococcus granulosus metacestode during infection of its intermediate host

Cystic hydatid disease (CHD) is caused by infection with the Echinococcus granulosus metacestode and affects both humans and livestock. In this work, we performed a proteomic analysis of the E. granulosus metacestode during infection of its intermediate bovine host. Parasite proteins were identified in different metacestode components (94 from protoscolex, 25 from germinal layer and 20 from hydatid cyst fluid), along with host proteins (58) that permeate into the hydatid cyst, providing new insights into host‐parasite interplay. E. granulosus and platyhelminth EST data allowed successful identification of proteins potentially involved in downregulation of host defenses, highlighting possible evasion mechanisms adopted by the parasite to establish infection. Several intracellular proteins were found in hydatid cyst fluid, revealing a set of newly identified proteins that were previously thought to be inaccessible for inducing or modulating the host immune response. Host proteins identified in association with the hydatid cyst suggest that the parasite may bind/adsorb host molecules with nutritional and/or immune evasion purposes, masking surface antigens or inhibiting important effector molecules of host immunity, such as complement components and calgranulin. Overall, our results provide valuable information on parasite survival strategies in the adverse host environment and on the molecular mechanisms underpinning CHD immunopathology.     

Comparative cytotoxicity of secondary hydatid cysts, protoscoleces, and in vitro developed microcysts of Echinococcus granulosus.

Infection with the metacestode of Echinococcus granulosus is characterized by a concomitant immunity. Survival of established and developing hydatid cysts in the intermediate host implies a mechanism to modulate its immunological reactions. In order to investigate this mechanism, secondary hydatid cysts were isolated from intraperitoneally infected laboratory white mice (strain NMRI) 12 months p.i. A number of hydatid cysts were freed from the surrounding host adventitial tissue. Monolayer cultures of non-stimulated peritoneal macrophages of NMRI mice were prepared and incubated in the presence of the hydatid cysts. By means of a trypan blue exclusion test and by measuring the incorporation of tritium labelled uridine, it was found that the presence of hydatid cysts reduced the viability of the macrophages in vitro. Toxic substances are probably secreted since the medium of cultured hydatid cysts also displayed cytotoxic activity. Hydatid cysts with adventitia, as well as culture medium of those cysts, were less toxic. When toxins, partially purified from hydatid cyst fluid, were previously incubated on a collagen coated surface, a reduced level of toxicity was found, suggesting that collagen of the host adventitia may play a role in controlling the liberation of toxins by the hydatid cyst. Virtually no toxicity was exerted by protoscoleces or by the medium of cultured protoscoleces, in contrast to in vitro vesiculated protoscoleces (so called microcysts). The results reveal a novel feature of hydatid cysts that may play a role in the survival of the parasite in the immunized host.     

Metalloproteinases in the larvae of Echinococcus granulosus.

Proteolytic activity in hydatid cyst fluid, cyst membranes and protoscoleces of E. granulosus was analyzed by electrophoresis in gelatin-containing polyacrylamide gels, including characterization with a set of protease inhibitors. All contained metalloproteinases in the range 60-120 kDa, with neutral/alkaline pH optima. Major activity was observed in hydatid fluid and the membranes (five bands) with both exhibiting similar electrophoretic patterns. The samples prepared from protoscoleces shared only some of these bands.     

Echinococcus granulosus: Permeability of hydatid cysts to mebendazole in mice

Mice infected with Echinococcus granulosus were given one dose of 15 mg kg^?1 of ¹⁴C-mebendazole by gavage. Blood, hydatid cyst fluid and membranes were collected and counted at varying intervals thereafter. Radioactivity in blood peaked by 16 h and declined rapidly thereafter. Activity in hydatid cyst fluid paralleled that in blood but in amounts of only 5–10%. While levels of radioactivity in hydatid cyst membranes for the most part paralleled those of blood, in several samples they remained stable or increased from 16 to 48 h while those in blood had decreased to baseline. Protoscolices lost all signs of viability by 48 h after treatment.     

Echinococcus granulosus: lethal effect of low voltage direct electric current on hydatid cyst protoscoleces

We have studied a small scale method for killing hydatid cyst protoscoleces using low voltage direct electric current. After collecting hydatid cysts from infected organs of slaughtered animals, protoscoleces were cultured in four different media: hydatid cyst fluid, RPMI, normal saline, and Tris buffer, respectively. Protoscoleces from each of the above media were then transferred to an electrolysis device through which different electric current densities were applied. For measuring the survival rate of protoscoleces, flame cell movement and eosin staining was used. The results show that the survival rate of protoscoleces in hydatid fluid was dependent on the electric current density and the time of the applied current. Current densities of 62.5 mA/cm2 (11 V), 53.71 mA/cm2 (10 V), and 18.18 mA/cm2 (5 V) after 1, 2, and 3 min, respectively, killed all the parasites in the hydatid fluid. However, a current density of 7 mA/cm2 (9 V) in RPMI medium after 3 min was most effective.     

Echinococcal cyst of the subcutaneous tissue: a rare case report

Hydatid disease is an endemic zoonosis caused by the cestode Echinococcus granulosus. The most common affected organs are the liver and the lungs. Echinococcal cyst located in the subcutaneous tissue constitutes an extremely rare manifestation of the hydatid disease. Here we present a case report with a cystic mass in the subcutaneous tissue of the left gluteal area, diagnosed as being a hydatid cyst.     

Respiratory and cardiovascular responses resulting from intravenous injection of sheep hydatid cyst fluid to dogs

Effects of intravenous injection of sheep hydatid cyst fluid on respiratory and cardiovascular systems were studied in sodium pentobarbital-anesthetized dogs. Under sterile conditions, the fluid was drawn from the liver and lung hydatid cysts of sheep, and centrifuged for 20 min at 500g. The supernatant fluid was used as the test solution. In a majority of animals, administration of 5 ml of the cyst fluid caused a sharp fall in arterial blood pressure, with or without respiratory changes. The latter, when present, included temporary cessation of respiration followed by rapid shallow breathing. There was usually an increase in the heart rate as the arterial blood pressure dropped. Atropine, 0.5 mg/kg body wt, given subcutaneously prior to the cyst fluid administration, did not block the responses. Cardiovascular responses produced by intravenous injection of cyst fluid were similar in dogs with positive pressure breathing and with spontaneous breathing. In 7 out of 10 responsive dogs, pretreatment with the antihistamine, chlorpheniramine, abolished the respiratory and cardiovascular responses.     

Cytodiagnosis of hydatid disease presenting with Horner's syndrome: a case report

BACKGROUND: The diagnosis of echinococcosis is based on clinical, radiologic and serologic findings. Fine needle aspiration (FNA) is useful in evaluating the lesion when the presentation is atypical. We report a case of hydatid cyst at the lung apex in which the diagnosis was made on FNA, with no adverse reaction to the aspiration. CASE: A 30-year-old male, a chronic smoker, presented with pain radiating to the medial two fingers of the right hand for two years. He also had miosis and ptosis of the right eye and anhidrosis of the right side of theface. There was wasting of small muscles in the right hand. Magnetic resonance imaging revealed a hyperintense mass at the apex of the right lung, thoracic inlet and adjacent vertebral bodies. A clinical diagnosis of Horner's syndrome due to pancoast tumor was rendered. Ultrasound-guided FNA showed protoscolices, scattered hooklets and bits of acellular laminated membrane, characteristic of echinococcosis. Surgical excision of the cyst was done. Gross and histopathologic examination of the excised cyst confirmed the diagnosis. CONCLUSION: FNA, though traditionally contraindicated, is a highly desirable, rapid, noninvasive diagnostic mode for echinococcosis. In view of its pathognomonic cytomorphologic features and numerous reports on cytodiagnosis in the literature, it is time to evaluate the diagnostic benefits and weigh the risks against the advantages of the technique.     

Primary Renal Hydatid Cyst: Mis-Interpretation as a Renal Malignancy

Primary renal echinococcosis, a rare disease involving the kidney, accounts for 2-3% of human echinococcosis. A 64-year-old female patient from Uzbekistan presented with complaints of left flank pain. A CT scan revealed a cystic mass in the upper to midpole of the left kidney. We regarded this lesion as a renal malignancy and hand-assisted laparoscopic radical nephrectomy was performed to remove the renal mass. The mass consisted of a large unilocular cyst and multiple smaller cysts without any grossly visible renal tissue. The final pathologic diagnosis was a renal hydatid cyst. For patients from endemic areas, hydatid cyst should be included in the differential diagnosis. Here, we present a case of renal hydatid cyst in a female patient who relocated from Uzbekistan to Korea.     

Serum Antigen and Antibody Detection in Echinococcosis: Application in Serodiagnosis of Human Hydatidosis

Diagnosis of hydatidosis is based on immunodiagnostic methods along with radiological and ultrasound examinations. The objectives of the present study were to develop a specific and simple antigen-based ELISA method for diagnosis of hydatidosis and compare it with antibody detection method. The subjects in this study included 89 patients in the following groups: surgically confirmed hydatidosis patients (35 cases), control with other parasitic diseases (29 cases), and healthy controls (25 cases). Hyperimmune serum was raised against hydatid cyst fluid in rabbits. Anti-hydatid cyst IgG was purified by affinity chromatography using protein A column and labeled with horseradish peroxidase. Collected sera were assessed for hydatid cyst antigens and antibody by ELISA. Circulating hydatid antigen was found in 9 out of 35 patients with surgically confirmed hydatidosis. A sensitivity of 25.7% and a specificity of 98.0% were calculated for the antigen detection assay. Antibody detection by indirect ELISA, using antigen B, showed that 94.2% of patients (33 cases) have anti-hydatid cyst antibodies in their serum while cross reaction was noted in a few of non-hydatidosis patients. A sensitivity of 94.2% and specificity of 81.6% were found for the antibody detection assay. Findings of this study indicated that antibody detection assay is a sensitive approach for diagnosis of hydatid cyst while antigen detection assay might be a useful approach for assessment of the efficacy of treatment especially after removal of the cyst.     

Does hydatid disease have protective effects against lung cancer?

We hypothesized that solid tumors rarely occur in patients with hydatid disease. We obtained the serum of 14 patients diagnosed with hydatid disease, the serum of 10 patients who did not have a history of hydatid disease, and the hydatid cyst fluid from six patients. These sera and fluid samples were added at different concentrations to NCI-H209/An1 human lung small cell carcinoma cells and L929 mouse fibroblasts as a control group. Sera of patients with hydatid diseases had cytotoxic effects on NCI-H209/An1 cells, but they did not have cytotoxic effects on fibroblast cells. Sera from healthy subjects did not have a cytotoxic effect on the tumor cell line or control fibroblasts. Cyst fluid, also, did not have toxic effects on the NCI-H209/An1 cell line, but was toxic to fibroblasts up to a 1:32 dilution. Sera from patients with hydatid disease had cytotoxic effects on human small cell lung cancer cells in vitro.     

Free amino acid concentration in hydatid cyst fluids from fertile and infertile human and animal Echinococcus granulosus.

The aim of this study was to search and compare free amino acid composition of fertile and infertile cyst fluids obtained from humans and animals infected naturally with Echinococcus granulosus, by using automated analysis based on cation-exchange chromatography with post-column ninhydrin derivatization system. 11 free amino acids from fertile (sheep origin), nine from infertile (cattle origin), 13 from infertile (human origin) hydatid cyst fluids and 19 amino acids from sera of patients with hydatid infection were detected. The levels of glycine, alanine, valine and lyrosine in fertile and infertile hydatid cysts fluids were significantly higher than in sera from patients with hydatid cysts. Glycine level in the fertile hydatid cyst fluids (sheep origin) was significantly higher than those of infertile cysts fluids (cattle and human origin) and sera with hydatid patients. Glycine level in fertile hydatid cyst fluids was about two times more concentrated in infertile cattle cyst fluids, 10 times more concentrated in infertile human hydatid cyst fluids and 13 times more concentrated in sera with hydatid patients. On the other hand, alanine and valine concentration in the fertile and infertile cyst fluids were at similar level with the exception that valine level in fertile cyst fluids was 12 times more concentrated in infertile human cyst fluids. The levels of tyrosine, citrulline, leucine, isoleucine and lysine amino acids in fertile and infertile hydatid cyst fluids were similar. Our findings with respect to fertile and infertile cysts fluids showed that free amino acids concentrations in cyst fluids were significantly, higher in sera from patients with hydatid cyst. Total amount of free amino acids content in fertile and infertile cyst fluids was three to eight times higher from that of human sera with hydatid patients.