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Enhanced larval cyst growth of Echinococcus multilocularis in praziquantel-treated jirds (Meriones unguiculatus).

Jirds (Meriones unguiculatus) inoculated intraperitoneally with cystic material of Echinococcus multilocularis were given daily oral treatments of praziquantel at 300 mg/kg of body weight (bw) or dimethyl sulfoxide vehicle for five-day treatment regimens starting at 29 days postinoculation (PI) up to 69 days PI. At 39 or 49 days PI, the growth of the larval cystic mass (LCM) in jirds following a single or two five-day treatment regimens was significantly enhanced (P < 0.05) by 129.0% (2.3-fold) or 102.9% (2.0-fold), respectively. At 59 or 69 days PI following three or four five-day treatments with praziquantel, LCM growth was enhanced by 47.8% (1.5-fold) and 44.1% (1.4-fold), respectively, but was no longer significantly different than that in control jirds. A single five-day treatment on 29-33 days PI (with necropsy at 69 days PI) significantly enhanced the growth of the LCM by 87.6% (1.9-fold). Parasites from praziquantel treatment regimens examined ultrastructurally showed consistent damage to the germinal membrane evidenced by vacuolization and rupture of syncytial cytoplasm, rupture and coalescence of the electron-lucent vesicles just below the microvilli of the tegumental surface, and swelling and rounding of mitochondria. At 39 days PI, increased blebbing of the germinal membrane into the lumen of the LCM in praziquantel-treated animals was observed by scanning electron microscopy. The treatment-induced blebs were identified as nucleated germinal cells by transmission electron microscopy and appeared to be responsible for metastasis and enhanced growth of the LCM. Although praziquantel damaged the ultrastructural integrity of the LCM, treatment failed to inhibit larval cyst growth or protoscolex development.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Seroepidemiologic screening of Echinococcus multilocularis infection in a European area endemic for alveolar echinococcosis.

In a serologic survey for Echinococcus multilocularis infection, we screened sera from 7,884 subjects from the Doubs Departement in France, an area endemic for alveolar echinococcosis (AE) of the liver. An enzyme-linked immunosorbent assay (ELISA) with a highly species-specific antigen (Em2) and an E. multilocularis crude antigen (Emc) was used for screening. An evaluation of the cost/benefit relationship of this screening, followed by therapeutic management of patients, was made and compared with the actual cost of the follow-up and treatment of the disease in symptomatic cases in this endemic area. Antibody reactions to Em2 and/or Emc made possible the detection of eight asymptomatic clinical cases (seroprevalence averaging 1/1,000), with typical lesions of active AE revealed by abdominal ultrasonography and computed tomography. All were seropositive using the Emc ELISA but two were seronegative using the Em2 ELISA. In five additional seropositive cases, the radiologic investigations revealed small calcified lesions similar to the lesions of abortive AE previously found in Alaska. The cost of this serologic screening program per screened subject and per diagnosed case averaged 50.00 French Francs (FF) (U.S. $8.60) and 60,000.00 FF (U.S. $10,909.00), respectively. The cost of diagnosis, follow-up and treatment of the patients was 5,086.00 FF (U.S. $929.00) per patient per month in the case of diseases diagnosed by the screening program and 7,086.00 FF (U.S. $1,288.00) per patient per month for patients with symptomatic AE. This survey indicates a high prevalence of AE in the target area; it confirms the long latency period of the larval growth in human AE and shows that abortive AE is present in Europe. The use of both the Emc and Em2 ELISAs seems to be better than using the Em2 ELISA alone. The cost of the hospitalization and treatment of the eight screened patients would appear to be relatively high. Even though two of them were asymptomatic, they had very severe forms of the disease. In fact, the total cost was much lower than the actual cost of the disease when diagnosed from clinical symptoms.

Adolescent↗

Second recorded case of human infection by Echinococcus oligarthrus.

The paper reviews a previously published case of hydatid disease in the human heart of a Brazilian person who died of tetanus. Based on present knowledge about the distinguishing characteristics of Echinococcus granulosus, E. vogeli, and E. oligarthrus, it was recognized that the infection was due to E. oligarthrus, mainly based the morphologic features of the hooklets of the protoscolex. This is the second human infection due to E. oligarthrus and the first showing wall features of cysts. Therefore, some human infections of polycystic hydatid disease observed outside the range of the bush dog, the only definitive host of E. vogeli (Panama to Northern Argentina), may be due to E. oligarthrus rather than to E. vogeli.

Aged↗

Increases in the effects of albendazole on Echinococcus multilocularis metacestodes by the dipeptide methyl ester (Phe-Phe-OMe).

Three months after infection with Echinococcus multilocularis, Mongolian gerbils were given either the dipeptide methyl ester (Phe-Phe-OMe) or a combination of Phe-Phe-OMe plus albendazole to treat alveolar echinococcosis. Each drug was given orally at the daily dose of 50 mg/kg of body weight following various administration regimens. Histologic and ultrastructural studies of parasites recovered from infected gerbil tissues showed that the dipeptide methyl ester increases the effect of albendazole.

Albendazole↗

Indication of the presence of two distinct strains of Echinococcus granulosus in Iran by mitochondrial DNA markers.

Sixteen isolates of Echinococcus granulosus, collected from Iranian patients at surgery, and from domestic animals, including sheep, goats, cattle, and camels at slaughterhouses in Tehran and central and southern Iran were analyzed for DNA nucleotide and predicted amino acid sequence variation within regions of the mitochondrial cytochrome c oxidase I (COI) and NADH dehydrogenase subunit I (NDI) genes. A polymerase chain reaction-restriction fragment length polymorphism method, based on the DNA sequence variation in the NDI gene, was also used to rapidly survey the E. granulosus isolates. The isolates were categorized into two distinct and uniform genotype groupings. The analysis clearly indicated that the camel/dog strain (G6 genotype) of E. granulosus as well as the cosmopolitan, common sheep strain (G1 genotype) occur in Iran. The G1 genotype was found present in all four human isolates examined and it was more prevalent in domestic animals than the camel-restricted G6 genotype. In E. granulosus-endemic areas of Iran it is evident, therefore, that the majority of E. granulosus-infected livestock animals can potentially act as reservoirs of human infection, and this has important implications for hydatid control and public health.

Amino Acid Sequence↗

Geographic information system-aided analysis of factors associated with the spatial distribution of Echinococcus multilocularis infections of foxes.

To investigate the influence of environmental factors on the spatial epidemiology of infections with Echinococcus multilocularis, foxes were sampled in a focal endemic region in the Northwest of Brandenburg, Germany, and examined for infection by the parasite. The locations where foxes were obtained were recorded in a geographic information system database. Positions of infected and uninfected foxes were analyzed on the background of geographic vector data of water, settlements, streets, forests, crop, and pasture. Fox positions were allocated to different land-use classes by use of a Landsat Thematic Mapper (TM) satellite image. Infected foxes were more frequently shot near water, in areas of high soil humidity, and on pastures, suggesting that dryness may limit the tenacity of E. multilocularis oncospheres. Thus open landscapes with humid soil seem to be favorable for the life cycle of the parasite. In contrast, infected foxes were significantly underrepresented in forest areas.

Animals↗

Short report: molecular genetic characterization of an unusually severe case of hydatid disease in Alaska caused by the cervid strain of Echinococcus granulosus.

Distinct Echinococcus granulosus life cycle patterns have been described in North America: domestic and sylvatic. Gene sequences of the sylvatic E. granulosus indicate that it represents a separate variant. Case-based data have suggested that the course of sylvatic disease is less severe than that of domestic disease, which led to the recommendation to treat cystic echinococcosis patients in the Arctic by careful medical management rather than by aggressive surgery. We recently reported the first two documented E. granulosus human cases in Alaska, with accompanying severe sequelae. Here we describe the results of molecular genetic analysis of the cyst material of one of the subjects that supported identification of the parasite as the sylvatic (cervid) strain and not the domestic (common sheep strain), which was initially thought to be implicated in these unusually severe Alaskan cases.

Animals↗

A genetic comparison of human and wildlife isolates of Echinococcus granulosus in Queensland: public health implications.

OBJECTIVE: To test the hypothesis that the hydatid parasite infecting macropods and dingoes in Queensland is a sylvatic strain of Echinococcus granulosus, distinct from the domestic strain which produces cysts in sheep and humans. DESIGN: Molecular biological techniques were used to compare DNA isolated from hydatid cysts from humans, local macropods and sheep from New South Wales and the United Kingdom, as well as from adult tapeworms in dingoes. SETTING: The human cysts were surgically resected from two patients seen with hydatidosis in Brisbane teaching hospitals over a one-year period. Neither patient had had previous contact with sheep farms. Macropods and dingoes were shot randomly in the localities where the patients presumably acquired their infections. Sheep liver cysts were obtained from abattoirs. METHODOLOGY: Studies comprised extraction of DNA from cysts, digestion by a series of restriction endonucleases, slab gel electrophoresis. Southern blotting and then hybridisation with defined DNA probes. Polymerase chain reaction, in combination with direct DNA sequencing, was used to compare DNA from cysts and adult worms from dingoes. RESULTS: The restriction fragment length polymorphism (RFLP) patterns of DNA from all cysts and a defined mitochondrial DNA sequence from all sources were indistinguishable. This finding is significant as both techniques can clearly distinguish between genetically distinct, well characterised strains of E. granulosus. CONCLUSIONS: Hydatid cysts are prevalent in some macropod populations and adult worms are common in dingoes. Since there are relatively few sheep-rearing areas in Queensland, contact with wild animals may be the main source of human hydatid infection in this State. The strain of E. granulosus in both patients was genetically indistinguishable from that found in macropods, dingoes and sheep from New South Wales and the United Kingdom. This strongly suggests that the domestic strain of E. granulosus, or a form very close genetically, freely infects Australian wildlife, and argues against the existence of a distinct sylvatic strain. The implications for public health are considerable.

Adult↗

Experimental evaluation of mink and Apodemus speciosus in the Echinococcus multilocularis life-cycle in Hokkaido, Japan.

The epizootiological status of mink (Mustela vison) as definitive hosts and Apodemus speciosus as intermediate hosts in the transmission of Echinococcus multilocularis in Hokkaido, Japan, were evaluated by orally inoculating mink with protoscoleces, and A. speciosus with eggs of the cestode, respectively. No tapeworms were recovered from the alimentary tract of the mink, and no hydatid cysts were recovered from the viscera of the egg-inoculated A. speciosus. We conclude that mink and A. speciosus cannot serve as definitive hosts and intermediate hosts of E. multilocularis, respectively, in Hokkaido.

Animals↗

Evaluation of fox-chasing enclosures as sites of potential introduction and establishment of Echinococcus multilocularis.

Following detection of Echinococcus multilocularis in red foxes (Vulpes vulpes) illegally imported into South Carolina (USA) for release in fox-chasing enclosures, a survey for E. multilocularis was conducted in four enclosures in Georgia (USA) and six enclosures in South Carolina. Survey methods included examination of potential small mammal intermediate hosts (n = 390) for E. multilocularis larvae, examination of fox and coyote (Canis latrans) scats (n = 59) for taeniid eggs, and examination of one possible canine definitive hosts for adult E. multilocularis. All intermediate and definitive hosts examined were negative for E. multilocularis and taeniid eggs were not recovered from fox and coyote fecal samples. Thus, E. multilocularis may not yet be established in fox-chasing enclosures in Georgia and South Carolina. Despite the failure to demonstrate E. multilocularis in the fox-chasing enclosures surveyed, translocation of wild canids from known enzootic regions should be discouraged because E. multilocularis is known to be ecologically adaptable and because contact with potentially infected definitive hosts during translocation is a public health risk.

Animals↗

New distribution records of Echinococcus multilocularis in the brown lemming from Barrow, Alaska, USA.

We identified Echinococcus multilocularis for the first time in brown lemmings (Lemmus trimucronatus) from Barrow, Alaska, USA. Of 467 brown lemmings trapped between 1995 and 2000, two males and two females (0.9%; 95% confidence interval=0.9+/-0.9%) were found to be infected with metacestodes of E. multilocularis. No metacestodes were found in 17 collared lemmings (Dicrostonyx rubricatus) also trapped at Barrow. In humans, E. multilocularis causes alveolar echinococcosis, which is potentially fatal. Knowledge of the distribution of this parasite is important to protect the public health.

Alaska↗

Effect of mebendazole and praziquantel on glucosephosphate isomerase and glyceraldehydephosphate dehydrogenase in Echinococcus granulosus cyst wall harbored in mice.

AIM: To study effects of antihydatid drugs on glucosephosphate isomerase (GPI) and glyceraldehydephosphate dehydrogenase (GAPDH) in Echinococcus granulosus cyst wall. METHODS: Mice infected with the parasite for 8-10 months were treated i.g. with mebendazole (Meb) or praziquantel (Pra). The activities of GPI and GAPDH in the cysts were measured by the formation of NADH or NADPH. RESULTS: GPI activity in the cyst wall was 197 +/- 103 U, while that of GAPDH was 25 +/- 13 U. When infected mice were treated i.g. with Meb 25-50 mg.kg-1.d-1 for 7-14 d, no apparent effect on the GAPDH activity in the cyst was found. In mice treated i.g. with praziquantel (Pra) 500 mg.kg-1.d-1 for 14 d, the GAPDH activity in the cyst wall was inhibited by 26.5%. As to GPI activity only the group treated i.g. with Meb 25 mg.kg-1.d-1 for 14 d showed 33.2% inhibition of the enzyme in the collapsed cyst wall. CONCLUSION: GPI and GAPDH are not the major targets attacked by the antihydatid drug.

Animals↗

Echinococcus multilocularis coproantigen detection by enzyme-linked immunosorbent assay in fox, dog, and cat populations.

A sandwich enzyme-linked immunosorbent assay (ELISA) for the detection of Echinococcus multilocularis coproantigens (EM-ELISA) was developed with polyclonal rabbit (solid phase) and chicken egg (catching) antibodies that were directed against E. multilocularis coproantigens and somatic worm antigens, respectively. In experimentally infected dogs and cats, coproantigens were first detectable 6-17 days postinfection (PI) in samples of 8 dogs (worm burdens at necropsy: 6,330-43,200) and from 11 days PI onward in samples of 5 cats infected with 20-6,833 worms. After anthelmintic treatment of 4 dogs and 5 cats at day 20 PI, coproantigen excretion disappeared within 3-5 days. The sensitivity of the ELISA was 83.6% in 55 foxes infected with 4-60,000 E. multilocularis, but reached 93.3% in the 45 foxes harboring more than 20 worms. The EM-ELISA was used in surveys of "normal" dog and cat populations in Switzerland. Among 660 dogs and 263 cats, 5 dogs and 2 cats exhibited a positive reaction. In 2 of these dogs (0.30%) and 1 cat (0.38%), intestinal E. multilocularis infections were confirmed by necropsy, polymerase chain reaction PCR, or both. The specificites of the ELISA in these groups were found to be 99.5% and 99.6%, respectively, if positive ELISA results that could not be confirmed by other methods were classified as "false positive" reactions.

Animals↗

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.

Animals↗

Evaluation of coproantigen diagnosis for natural Echinococcus multilocularis infection in red foxes.

The validity of a coproantigen ELISA for Echinococcus multilocularis was evaluated by comparison of three diagnostic methods; autopsy, egg examination and the ELISA. Of 71 foxes, 39 were found to be infected with the cestode at autopsy. The overall mean of worm burdens was 3,451, but the number varied (1-34,522). The ELISA could detect 94.9% (37/39) of the worm positives and there were no false-positives. Two false-negatives were infected with 1 and 4 cestodes, whereas 3 cases with similar worm burdens (2, 4 and 6 worms) were diagnosed as positives. This indicates the detection limit of the assay may be equivalent to less than 10 (in the worm burden). On the other hand, egg examination showed low sensitivity (43.6%, 17/39). These results suggest the ELISA has a potential to replace for the conventional methods.

Animals↗

Dose dependency of prednisolone on the establishment of Echinococcus multilocularis infection in an alternative definitive host, Mongolian gerbil.

This study revealed the dose dependency of prednisolone tertiary-butylacetate (PTBA) treatment on the establishment of Echinococcus multilocularis in the small intestine of Mongolian gerbil (Meriones unguiculatus) and that some of the physiological parameters of host were correlated with the doses of PTBA and establishment of the worm. Twenty Mongolian gerbils were divided into 5 groups, according to the doses of PTBA; 0 mg, 0.5 mg, 2 mg, 5 mg and 10 mg per head. All animals were injected intraperitoneally with PTBA every other day from 6 days before to 6 days after infection. Doses of PTBA and the number of worms recovered at 7 days post-infection showed a positive correlation (r = 0.929, P < 0.0001). The increase of total protein (TP) and the decrease of the percentage of lymphocytes in the peripheral leukocytes were dependent on doses of PTBA (TP: r = 0.811, P < 0.0001, percentage of lymphocyte: r = -0.92, P < 0.0001). The TP and the percentage of lymphocyte also correlated with the number of worms recovered (TP: r = 0.617, P = 0.0049; percentage of lymphocyte: r = -0.800, P < 0.0001).

Animals↗

Molecular evidence for the presence of a G7 genotype of Echinococcus granulosus in Slovakia.

Variability in Echinococcus granulosus is very important epidemiologically since strain characteristics may influence local patterns of transmission of hydatid disease. To classify the genotype presented in pig protoscoleces of the Slovak territory, a DNA-based approach has been used. Nucleotide sequences for a 471 bp region of the mitochondrial NADH dehydrogenase 1 (ND1) gene revealed a substantial affinity of isolates examined to the G7 genotype. Only a 0.9-3.4% sequence variation was recorded for E. granulosus samples compared with the reference G7 variant. To distinguish between G7 and G9 genotypes not differing in ND1 sequences, isolates were additionally examined by PCR-RFLP analysis of the nuclear ITS1 region. The resulting two-banded pattern is characteristic for the G7 strain. The data presented thus provides the first explicit evidence of the G7 genotype in the Slovak region.

Animals↗

[Echinococcus multifocal. Epidemiology, clinical findings and treatment].

Infection with the fox dwarf tapeworm, Echinococcus multilocularis, has for the first time been found in a fox in Denmark, and E. multilocularis infection is therefore now a possible diagnosis in Danish patients. This paper reviews the life cycle, epidemiology, clinical symptoms, diagnosis, and treatment of E. multilocularis infections.

Animals↗