Erroneous reports of the neotropical Echinococcus oligarthrus as a cause of echinococcosis in India.

Two cases of submandibular echinococcosis have been reported in adult residents of India (Kini et al., 1997; Sahni et al., 2000). It was stated in both publications that the cestode was E. oligarthrus, "a rare variety of E. granulosus", based on morphogical characteristics. It was also mentioned that 3 cases of E. granulosus with similar submandibular metacestode had been previously diagnosed in India. The metacestode is readily distinguishable from that of E. oligarthrus. The final hosts of E. oligarthrus are several neotropical species of wild cats and metacestode occurs in hystricognath, all of them absent in India. The larval stage of all 4 species of Echinococcus can be differentiated on the basis of the form and dimensions of rostellar hooks of protoescoleces. With the information provided in the two papers we concluded that the 2 cases of echinococcosis described in India were due to E. granulosus.

Cystic echinococcosis in the Arctic and Sub-Arctic.

The northern biotype of Echinococcus granulosus occurs throughout the holarctic zones of tundra and taiga, from eastern Fennoscandia to the Bering Strait in Eurasia and in North America from arctic Alaska approximately to the northern border of the United States. The cycle of the cestode is complex in taiga at lower latitudes, because of the greater diversity of potential hosts. In the Arctic and Subarctic, however, four patterns of predator/prey relationships may be discerned. Two natural cycles involve the wolf and wild reindeer and the wolf and elk (moose), respectively. Where deer of the two species coexist, both are prey of the wolf; the interactions of the wolf and elk are here described on the basis of long-term observations made on Isle Royale (in Lake Superior near the southern limit of taiga), where only the wolf and elk serve as hosts for E. granulosus. A synanthropic cycle involving herding-dogs and domesticated reindeer caused hyperendemicity of cystic echinococcosis in arctic Eurasia, mainly in northeastern Siberia. The 4th pattern, a semi-synanthropic cycle, formerly existed in Alaska, wherein sled-dogs of the indigenous hunters became infected by consuming the lungs of wild reindeer. The sequence of changes in life-style inherent in the process of acculturation affected the occurrence of cystic echinococcosis among nomadic Iñupiat in arctic Alaska. When those people became sedentary, the environs of their early villages soon became severely contaminated by faeces of dogs, and cases of cystic echinococcosis occurred. Compared to cystic echinococcosis caused by E. granulosus adapted to synanthropic hosts (dog and domestic ungulates), the infection produced by the northern biotype is relatively benign. Nearly all diagnosed cases of cystic echinococcosis (> 300) in Alaska have occurred in indigenous people; only one fatality has been recorded (in a non-indigenous person). After sled-dogs were replaced by machines, cases have become rare in Alaska. A similar effect has been observed in Fennoscandia, in the Saami and domesticated reindeer. Recent records indicate that the prevalence of cystic echinococcosis is increasing in Russia, suggesting that dogs are used there in herding.

Characterization of the laminated layer of in vitro cultivated Echinococcus vogeli metacestodes.

The metacestode (larval) stages of the cestode parasites Echinococcus vogeli and E. multilocularis were isolated from the peritoneal cavity of experimentally infected C57BL/6 mice and were cultured in vitro for a period of up to 4 mo under conditions normally applied for the in vitro cultivation of E. multilocularis metacestodes. In contrast to E. multilocularis, E. vogeli did not exhibit extensive exogenous budding and proliferation but increased in size with a final diameter of up to 10 mm. Most metacestodes contained protoscoleces, singly or in groups, either associated with brood capsules or growing directly out of the germinal layer. Each individual metacestode was covered by an acellular translucent laminated layer that was considerably thicker than the laminated layer of E. multilocularis metacestodes. The ultrastructural characteristics, protein content, and carbohydrate composition of the laminated layer of in vitro cultivated E. vogeli and E. multilocularis were assessed using transmission electron microscopy, lectin fluorescence labeling, and lectin blotting assays. The laminated layer of E. vogeli is, as previously described for E. multilocularis metacestodes, largely composed of N-acetyl-beta-D-galactosaminyl residues and alpha- and beta-D-galactosyl residues, as well as of the core structure of O-linked carbohydrate chains, N-acetylgalactosamine-beta-1,3-galactose. However, in contrast to E. multilocularis, N-linked glycopeptides and alpha-D-mannosyl and/or glucosyl residues were also associated with the laminated layer of E. vogeli. The laminated layer from both species was isolated from in vitro cultivated metacestodes, and the purified fractions were comparatively analyzed. The protein:carbohydrate ratio (1:1) was similar in both parasites; however, the protein banding pattern obtained by silver staining following sodium dodecyl sulfate polyacrylamide gel electrophoresis suggested intrinsic differences in protein composition. A polyclonal antiserum raised against the E. multilocularis laminated layer and a monoclonal antibody, G11, directed against the major E. multilocularis laminated layer antigen Em2 did not cross-react with E. vogeli, indicating distinct compositional and antigenic differences between these 2 parasites.

A phylogenetic hypothesis for species of the genus Taenia (Eucestoda : Taeniidae).

Cladistic analysis of a numerical data matrix describing 27 characters for species of Taenia resulted in 4 most parsimonious phylogenetic trees (174 steps; consistency index = 0.28; homoplasy index = 0.72; retention index = 0.48). Monophyly for Taenia is diagnosed by the metacestode that is either a cysticercus or a form derived from a bladder-like larva; no other unequivocal synapomorphies are evident. Tree structure provides no support for recognition of a diversity of tribes or genera within the Taeniinae: Fimbriotaeniini and Taeniini have no phylogenetic basis. Hydatigera, Fimbriotaenia, Fossor, Monordotaenia, Multiceps, Taeniarhynchus, Tetratirotaenia must be subsumed within Taenia as synonyms. Taenia saginata and Taenia asiatica are sister species and distantly related to Taenia solium. Cospeciation with respect to carnivorous definitive hosts and Taenia appears to be limited. Although felids are putative ancestral hosts, contemporary associations appear to have resulted from extensive host-switching among felids, canids, hyaenids, and others. In contrast, relationships with herbivorous intermediate hosts are indicative of more pervasive coevolution; rodents as intermediate hosts are postulated as ancestral for the Taeniidae, Taenia + Echinococcus. Patterns appear consistent with rapid shifts between phylogenetically unrelated carnivores but among those that historically exploited a common prey resource within communities in specific biogeographic regions.

Histogenesis in the metacestode of Echinococcus vogeli and mechanism of pathogenesis in polycystic hydatid disease.

Histogenesis of the metacestode of Echinococcus vogeli was traced mainly in rodents inoculated intraperitoneally with finely minced infective vesicles. The fragments aggregated in the peritoneal cavity and coalesced, forming structures (plaques) from which primary vesicles arose. From primordia in their germinal tissue, exogenous vesicles developed, enlarged, and migrated outward to the surface of the laminated membrane, where they remained attached and proliferated. Each unit of vesicles so formed retained discrete identity and, within 6-8 mo, acquired an adventitia; thereafter, exogenous multiplication ceased and endogenous proliferation supervened. Large numbers of daughter cysts arose in the germinal tissue lining chambers within the units; endogenous proliferation also finally ceased, and the daughter cysts produced brood capsules containing protoscoleces. Primordia of exogenous vesicles were not observed in the walls of daughter cysts. Production of protoscoleces involved 3 processes: they developed in typical brood capsules, singly in minute brood capsules, or directly from germinal tissue. Exogenous proliferation is not characteristic in the natural intermediate host of E. vogeli, the paca. Evidently in primates, the initial proliferation in the liver is followed by extension of the metacestode into the peritoneal cavity and eventual invasion of abdominal and thoracic organs. Exogenous proliferation by a process unique to E. vogeli accounts for the clinical course of polycystic hydatid disease.

Immunodiagnosis of polycystic hydatid disease/polycystic echinococcosis due to Echinococcus vogeli.

A crude antigenic metacestode extract from Echinococcus vogeli was assessed by enzyme-linked immunosorbent assay (ELISA) and showed strong binding activity with serum antibodies from patients with polycystic echinococcosis. Major cross-reactions occurred with serum antibodies from patients with cystic and alveolar echinococcosis and from patients infected with other species of helminths. An E. vogeli antigen fraction, Ev2, was subsequently purified by immunosorption. The respective Ev2 ELISA demonstrated improved specificity, allowing discrimination of non-Echinococcus infections from polycystic echinococcosis. Based upon the calculation of a comparative (Ev-crude ELISA versus Ev2 ELISA) reactivity index, it became possible to discriminate all cystic echinococcosis cases, but only some alveolar echinococcosis cases, from polycystic echinococcosis. Immunoblot analyses revealed an antibody banding pattern highly conserved among polycystic, cystic, and alveolar echinococcosis. However, immunoblotting reliably distinguished between echinococcosis and all non-Echinococcus infections.

Alveolar hydatid disease. Review of the surgical experience in 42 cases of active disease among Alaskan Eskimos.

OBJECTIVE: The authors reviewed the pathophysiology and clinical management of endemic alveolar hydatid disease in Alaskan Eskimos, incorporating recent developments in diagnosis and treatment. SUMMARY BACKGROUND DATA: Alveolar hydatid disease is a highly lethal zoonotic infection caused by the larval stage of Echinococcus multilocularis. This cestode is restricted geographically to northern climates, where foxes and small rodents represent the natural hosts. Domestic dogs also may serve as definitive hosts, and thus, transmit the parasite to humans. Human infection is characterized by the development of a cancer-like hepatic mass, which may extend to adjacent structures or metastasize to distant sites. If the infection goes untreated, mortality reaches 80%. METHODS: The medical records of all patients with alveolar hydatid disease diagnosed or treated at the Alaska Native Medical Center between 1951 and 1993 were reviewed. Forty-two cases of active disease are presented. RESULTS: Nine patients underwent resection of hepatic lesions with intent to cure, and each had a favorable result. Average post-diagnosis survival of those patients was 22 years; six still are living and free of disease. Partial resections or drainage procedures were performed in ten patients. Chemotherapy was used to augment the surgical treatment of eight patients, and four received chemotherapy alone, resulting in improved outcomes compared with historic controls. Late complications included hepatic abscess, biliary obstruction, and portal venous hypertension. CONCLUSIONS: Whereas alveolar hydatid disease rarely is encountered in other areas of North America, the biologic potential for spread of the disease may be increasing because of illegal importation of infected foxes to the Eastern seaboard. Therefore, the surgical community should maintain an awareness of the diagnosis and management of this potentially devastating parasitic infection.

Parasiticidal effect of chemotherapy in alveolar hydatid disease: review of experience with mebendazole and albendazole in Alaskan Eskimos.

Evidence that the larval stage of Echinococcus multilocularis in humans is killed by chemotherapy is presented in a review of our 17-year experience with treatment of alveolar hydatid disease in Alaska. The efficacy of chemotherapy was assessed with use of an in vivo assay of parasite viability by means of inoculation of voles, immunohistochemical tests, and histopathologic findings. Of 14 tests performed for nine patients, 12 in vivo assays (86%) were negative after chemotherapy, while only two (17%) of 12 vole tests for seven untreated patients were negative. Regression of arrest of growth of metastatic and primary hepatic lesions, together with their partial-to-complete calcification and prolonged survival times has been observed among patients treated with the benzimidazole compounds. For six who received appropriate chemotherapy, treatment has been discontinued for an average of 4.6 years (range, 3-7 years) without an increase lesion size or other evidence of reactivation.

Cysticerci (Cestoda: Taeniidae) from white-tailed deer, Odocoileus virginianus, in southern Florida.

Metacestodes (cysticerci) of Taenia omissa Lühe, 1910, and Taenia hydatigena Pallas, 1776, were found in 9 and 1 of 124 white-tailed deer, respectively, in southern Florida in 1984-1986. Intensities of T. omissa varied from 1 to 15 (mean = 4.6); only 1 cysticercus of T. hydatigena was collected. No significant difference in the prevalences of T. omissa according to sex, age, or locality was observed.

A programme to reduce the risk of infection by Echinococcus multilocularis: the use of praziquantel to control the cestode in a village in the hyperendemic region of Alaska.

This paper reports the results of a 10-year field trial designed to reduce the risk of infection by Echinococcus multilocularis to residents of a village in a hyperendemic area (Savoonga, St. Lawrence Island). The objective was to prevent dispersal of infective eggs of the cestode within the village by means of monthly treatments of dogs with praziquantel. Northern voles, Microtus oeconomus, present in the village as commensals, served as an index of risk, as the incidence of infection in the voles provides information about the availability of eggs within the confines of the inhabited area. Voles were examined annually during early June before the population of overwintering voles was diluted by the first annual litters. The pretreatment infection-rate within the village was 29% (range 22-35%), and in control areas at some distance from the village for the entire study period it averaged 53% (284 infected voles from a sample of 533). Some fluctuation in incidence of infection in village voles occurred, apparently depending on the extent to which the residents kept their dogs chained and thus available for treatment. The success of the programme was demonstrated by the reduction in prevalence of infection to about 1% of voles in 1985, and an average infection rate during the last five years of the study of 5% (29 infected voles in a sample of 582). This 83% average reduction in the prevalence of the larval cestode within the village reflects a corresponding reduction in the risk of acquiring by the residents of the village. The method would be applicable for the control of E. multilocularis in most hyperendemic regions. Success depends, however, on elimination of unrestrained dogs and a precise schedule of treatment.

Cysticerci of Taenia mustelae in the fox squirrel.

The liver of a fox squirrel (Sciurus niger rufiventer) contained many clear, spherical cysts approximately 1 mm in diameter throughout the parenchyma. On dissection, most of these exhibited a dense, white area with four muscular suckers at a single point on the bladder. Based on the size of the organisms, the characteristically tiny hooks on the scolex and the location of the cysts in this host, the parasites were identified as cysticerci of Taenia mustelae. This is the first report in this host. The parasites were surrounded by an intense inflammatory response consisting primarily of lymphocytes mixed with some eosinophils, and early deposition of fibrous connective tissue.

Growth of a Japanese isolate of Taenia crassiceps in intermediate and definitive hosts.

Higher infection rates were observed in gerbils and voles than in ICR mice after oral inoculation with eggs of a Japanese isolate of Taenia crassiceps. Asexual reproduction of T. crassiceps cysticerci was observed in all gerbils and voles infected i.p. with the cysticerci. However, ICR mice and Wistar rats were not suitable for the asexual proliferation of T. crassiceps. The hooks of cysticerci from mice were smaller than those from gerbils. In experimentally infected puppies, parasite development was noted as follows: strobilation and initial differentiation of the genital primordia on day 7 postinoculation (p.i.), appearance of the testes on day 9, observation of the ovaries on day 10, and development of the lateral branches of the uterus on day 15. The prepatent period was 27-31 days. After day 15 p.i., most of the worms were recovered from the middle third of the small intestine. The number of proglottids shed per day by each strobila was about 1. The number of eggs contained in a gravid segment was about 13,000.

Echinococcus vogeli Rausch and Bernstein, 1972, from the paca, Cuniculus paca L. (Rodentia: Dasyproctidae), in the Departamento de Santa Cruz, Bolivia.

Among approximately 2,000 mammals examined for helminths in various regions of Bolivia during 1983-1987, cysts of Echinococcus vogeli Rausch and Bernstein, 1972, were found in a single paca, Cuniculus paca L., collected at La Laguna, Departamento de Santa Cruz (lat. 16 degrees 36'W; long. 62 degrees 42'S). This record, the first from Bolivia, represents a considerable extension of the known geographic range of this species in South America. Upon analysis of the morphologic characteristics of the protoscoleces derived from the cysts, the sizes of rostellar hooks from the material from the paca were found to be well within the ranges reported in previous studies. Statistical analysis of frequency distributions of hook characteristics revealed some deviations from normality. These results indicate that parametric statistics should be applied with caution in analyses of inter-and intraspecific variation of morphologic characteristics of hooks of metacestodes of the genus Echinococcus.

Postoncospheral development and cycle of Taenia polyacantha Leuckart, 1856 (Cestoda: Taeniidae). First part.

The postoncospheral development and cycle of Taenia polyacantha Leuckart, 1856, an holarctic species of cestode, were investigated in the laboratory as well as in the tundra of northern Alaska. Foxes, Alopex lapogus (L.) and Vulpes vulpes (L.), serve as final host of T. polyacantha; the northern vole, Microtus oeconomus (Pallas), and the brown lemming, Lemmus sibiricus (Kerr), are important as the intermediate host. As determined in experimentally infected voles and lemmings, the oncosphere of T. polyacantha transformed to a primary vesicle in the liver. On the 6th day postexposure, coinciding with its migration to the peritoneal cavity, the larval cestode consisted of a minute aggregation of secondary vesicles. By 9-10 days postexposure, the secondary vesicles dissociated, thereafter developing independently to infective cysticerci by 30-40 days postexposure. At an age of about 60 days, the infective larvae began to undergo further growth and morphological modification, which led to acquisition of some strobilar characteristics by the forebody. Such late transformation of a cysticercus to a more advanced form of larva is known otherwise only in Taenia martis (Zeder, 1803). Differences in numbers and sizes of rostellar hooks provided the basis for recognition of two taxa at the infraspecific level: Taenia p. polyacantha Leuckart, 1856, distributed in Eurasia to the south of the zone of tundra, and T. p. arctica ssp. nov., present throughout the holarctic tundra. Observations concerning interactions of T. polyacantha and its hosts are discussed.

Application of the avidin-biotin immunohistochemical method for the diagnosis of alveolar hydatid disease from tissue sections.

The avidin-biotin immunohistochemical (ABC) method was applied to the diagnosis of alveolar hydatid disease in patients from the hyperendemic region of western Alaska. The test was specific for larval cestodes of the genus Echinococcus, and exhibited high sensitivity. A negative reaction was obtained in all cases in which spontaneous death of the larval cestode had occurred. Results were inconsistent in sections of lesions from patients who had received long-term therapy with mebendazole, because of the persistence of antigens for a considerable time following the death of the parasite.

Demonstration of amyloid and immune complex deposits in renal and hepatic parenchyma of Alaskan alveolar hydatid disease patients.

Histopathological changes and the incidence of amyloid and immune complex (IC) deposits were examined in six liver (autopsy or biopsy) and two kidney (autopsy) samples from seven Alaskan alveolar hydatid disease (AHD) patients. Both the kidney samples showed evidence of severe nephropathy and glomerular IC deposits. Two of the six liver samples had typical amyloid deposits. All the liver samples examined showed variable amounts of parenchymal atrophy adjacent to the locus of larval Echinococcus multilocularis, periportal and portal fibrosis. Thus secondary amyloidosis and IC-mediated pathological changes should be included as possible complications of AHD.

Albendazole therapy in alveolar hydatid disease: a report of favorable results in two patients after short-term therapy.

Albendazole was administered preoperatively to two patients with active alveolar hydatid disease for 58 and 84 days. Vesicles of larval Echinococcus multilocularis obtained from surgical tissues were inoculated into red-backed voles for in vivo testing viability. No proliferation of the larval cestode had occurred when the animals were dissected three months post-inoculation. These findings suggest that short-term therapy with albendazole was effective in killing the larval cestode in these two cases. Albendazole was found to be hepatotoxic but resulting transaminase abnormalities have been reversible. Close monitoring of liver function and hematology is essential in patients under albendazole therapy.

Spontaneous death of Echinococcus multilocularis: cases diagnosed serologically (by Em2 ELISA) and clinical significance.

Nine cases of asymptomatic alveolar hydatid disease (AHD) of the liver were diagnosed in 1985 among Eskimos from the endemic region of western Alaska. The patients were identified by screening with the enzyme-linked immunosorbent assay, using purified Echinococcus multilocularis antigen (Em2 ELISA). Five patients, and one diagnosed earlier (1979), were found to have lesions in which the larval E. multilocularis had died spontaneously at an early stage of infection. Viability was assessed histologically; by the avidin-biotin immunohistochemical method; and in vivo through intraperitoneal inoculation of membranes of the larval cestode into red-backed voles, Clethrionomys rutilus. The results were in agreement with the clinical impression, based on findings by computerized tomography and ultrasound scanning, and on the macroscopic appearance of the lesions, that the cestode was dead. Spontaneous death of E. multilocularis in humans has not been previously reported. The findings show that the Em2 ELISA may be positive in patients having lesions of AHD in which the etiologic agent is no longer viable.

Cysts resembling hydatids in a chicken.

Two large cysts recovered from the abdominal cavity of a domestic chicken superficially resembled hydatid cysts of Echinococcus spp. No protoscolices were present. Microscopic examination of the internal lining of one of the cysts revealed a single cuboidal to columnar, ciliated epithelium, leading to a diagnosis of oviduct cysts. Earlier records of hydatids of fowl are reviewed.

Consequences of continuous mebendazole therapy in alveolar hydatid disease--with a summary of a ten-year clinical trial.

The assessment of a ten-year clinical trial of continuous therapy in eight patients revealed further evidence of a significant therapeutic effect of mebendazole on alveolar hydatid disease. Life-expectancy was increased when compared to untreated historical controls, especially in the patients over 55 years of age. All symptomatic patients showed subjective improvement. In four patients, three had a 50% or greater reduction in the diameter of massive hepatic lesions, and in the fourth, progressively enlarging metastases were arrested. Fall in the IHA titre suggested that the causative organism had been destroyed in two additional patients. Of greater significance was the absence of progression of the disease process as measured by changes in the size of the hepatic lesion or lack of development of distant metastases in patients under therapy. In contrast, progressive enlargement of hepatic lesions or the appearance of distant metastases were cardinal features of untreated cases (15 of the 16 cases followed). In vivo determination of viability of tissues of the larval Echinococcus multilocularis from patients receiving long-term therapy was considered important in evaluating efficacy of the drug. Such tissues, obtained by autopsy from two patients under continuous therapy for four and ten years, failed to proliferate when inoculated into rodents (red-backed voles), whereas similar inoculations from untreated patients or those receiving 15 months' or less of therapy brought about production of vesicles in rodents in eight of 11 tests (73%). These two deaths, unrelated to therapy, resulted from late fibrotic constriction of end-stage parasitic lesions about the portal vein and major bile ducts. The clinical findings in combination with negative in vivo tests and other data indicate that the mebendazole therapy significantly alters the clinical course of alveolar hydatid disease. The evidence strongly indicates that long-term therapy may eventually have a lethal effect on the larval cestode in advanced disease.