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PubMed1986

Praziquantel adversely affects protoscoleces of Echinococcus granulosus in vitro.

Praziquantel was shown to have adverse effects on protoscoleces of Echinococcus granulosus in vitro. Exposure to one dose of praziquantel at a concentration of 10 micrograms/ml caused protoscoleces to die within 12 to 15 days. Protoscolecidal effects were more marked when cultures were exposed to the drug continuously by the addition of multiple doses. On the basis of these observations, there is a need to reappraise the metacestocidal potential of praziquantel, particularly with regard to the development of new drug regimes employing multiple dose or sustained release schedules. Possible reasons for the reduced susceptibility of protoscoleces and juvenile worms to praziquantel observed in this study are discussed.

Animals↗
PubMed1986

Echinococcus granulosus infection in dogs in Tambool, Sudan.

In the Tambool area (Central eastern Sudan) a survey was made of the infection rate of Echinococcus granulosus in dogs. From the total of 49 dogs shot by the police authority, 25 (51%) had E. granulosus in their intestines. The range of worm recovery was 7 to 28,400. These high infection and recovery rates could be explained by the rate of infection in the intermediate host (camel) slaughtered in the area.

Age Factors↗
PubMed1987

A comparison of two procedures for labelling the surface of the hydatid disease organism, Echinococcus granulosus, with 125I.

Living, intact protoscoleces of the British horse and sheep strains of Echinococcus granulosus were subjected to surface radioiodination procedures using 125I and Iodogen and 125I-Bolton Hunter reagent. Subsequent combined electron microscopy and autoradiography revealed specific surface membrane labelling with the Iodogen procedure, but significant tegumental labelling with the Bolton-Hunter reagent. The two parasite strains yielded different profiles of electrophoretically separated labelled proteins; the Iodogen method, not surprisingly, resulted in a less complex pattern of labelled polypeptides than the Bolton and Hunter reagent.

Animals↗
PubMed1988

Determination of the minimum time of praziquantel therapy required for the in vitro treatment of protoscoleces of Echinococcus granulosus.

Ovine and equine protoscoleces of Echinococcus granulosus were cultured for 26 days with our without praziquantel and viability assessed, by eosin exclusion, for cultures in various drug concentrations (50, 250 and 500 micrograms/l) and periods of exposure (1, 3 or 7 days (d] before removing/'rescuing' to drug-free medium. Drug efficacy was proportional to drug concentration and to length of exposure. At higher drug concentrations shorter exposures were required to produce the effect of continuous drug treatment, 1d therapy at 500 micrograms/l killing 96% ovine protoscoleces by day 14 whereas 7d therapy at 50 micrograms/l was required to produce a similar effect. Equine protoscoleces appeared marginally less susceptible than those of ovine origin. The relevance of the results in the need for peri-operative prophylaxis against spilled protoscoleces in man is discussed.

Animals↗
PubMed1990

Echinococcus granulosus: development of resistance to albendazole in an animal model.

Gerbils with well developed peritoneal cysts of Echinococcus granulosus were randomized to albendazole 50 mg/kg/day or untreated control. Treated animals had less disease at post mortem after 3 months of treatment. Cysts were then taken from both albendazole-treated and control animals and cultured in vitro either with or without albendazole sulphoxide (Alb Sx) 500 micrograms/L for 14 days. Viability of cysts was then established by implantation of whole cysts into gerbils. Whilst naive cysts were affected by Alb Sx (only 2 cysts developed/gerbil) cysts from animals treated with albendazole were not sensitive to further therapy (6.4 cysts/gerbil).

Albendazole↗
PubMed1990

An experimental in vitro model for evaluating drugs against protoscoleces of Echinococcus granulosus.

Pharmacological studies carried out on protoscoleces in vitro to standardize conditions that would permit a preliminary estimate of the efficacy of drugs with potential activity against Echinococcus granulosus are reported. Media such as PBS and Hanks solution, maintenance temperature, different pH values and concentrations of various solvents have been tested to check the effects on protoscolex survival in tubes in vitro. Mebendazole has been used as the pharmacological standard reference. Changes in the viability of protoscoleces have been used to demonstrate pharmacological activity. Best conditions were obtained employing Hanks solution and propylene glycol at low concentrations. Mebendazole was not completely effective at the concentrations achievable in human therapy. Linear, reproducible results demonstrated that Hanks solution provides an ideal medium for pharmacological studies. Among tested solvents, propylene glycol and dimethyl sulphoxide showed no lethal activity at low concentrations. At concentrations similar to those normally obtained in human sera, mebendazole, as in vivo, demonstrated only partial lethality for protoscoleces. The present study represents a new experimental approach to chemotherapy of hydatid disease.

Animals↗
PubMed1992

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.

Animals↗
PubMed1993

Distribution of some elements in hydatid cysts of Echinococcus granulosus from buffalo (Bubalus bubalis).

Distribution of some elements during development of endogenous daughter (hydatid) cysts of Echinococcus granulosus of buffalo origin was determined by atomic absorption spectrophotometry. These analyses showed that the distribution of copper and cobalt was highest in the smallest cysts. These elements gradually decreased in the cysts as they enlarged. The distribution of zinc, iron and manganese was very high in the smallest cysts in comparison to copper and cobalt, but these three elements decreased greatly in larger cysts. Iron and manganese were the only two elements found in very high concentrations in "thin-walled" cysts. Like all other elements both calcium and magnesium decreased as the cysts increased in size. In the thin-walled cysts magnesium increased by four times initially and this was followed by a two-fold "influx" of calcium in the "white-spots" in the above cysts. The significance of these findings is discussed in relation to earlier works.

Animals↗
PubMed1995

Echinococcus granulosus: observations on strobilar development in in vitro monophasic culture.

The in vitro cultivation technique of Echinococcus granulosus protoscoleces usually states the necessity of a biphasic medium with a solid protein substrate for strobilar development to take place; otherwise, in a monophasic medium, protoscoleces follow a vesicular development. However, in some monophasic cultures, the development of several strobilate individuals (in different quantities and stages of development, depending on the culture) were observed. The only known difference from cultures made previously and since, where the development was vesicular, was the batch of foetal calf serum used in the constitution of the liquid medium, and this is presumed to be the cause of this unexpected strobilar development.

Animals↗
PubMed1994

Antibody responses in the wild vole, Clethrionomys rufocanus bedfordiae, naturally infected with Echinococcus multilocularis by western blotting.

Antibody responses against crude antigens and the two (Em18 and Em16) epitopes of Echinococcus multilocularis were analysed by Western blotting using sera from the wild vole, Clethrionomys rufocanus bedfordiae (Crb), which were captured in Hokkaido, Japan and found to have been naturally infected with eggs, and compared with those in patients of AHD and mice experimentally infected with protoscoleces of this parasite. Antibody responses in the wild were demonstrated most clearly with anti-Crb-IgG antiserum but more faintly with anti-rat-IgG antiserum and poorly with anti-mouse-IgG or anti-human-IgG antisera or Protein G. Although only two serum samples of the wild vole found naturally infected were analysed, antibody responses against Em18 and Em16 in these voles appeared to be similar to those in AHD patients but differed from those in mice.

Animals↗
PubMed1996

An improved test system for PCR-based specific detection of Echinococcus multilocularis eggs.

For the sensitive detection of eggs of Echinococcus multilocularis in fox faeces by PCR we have evaluated a method based on the previous concentration of helminth eggs by a combination of sequential sieving of faecal samples and flotation of the eggs in zinc chloride solution. The eggs were microscopically detected in the fractions retained in 40 and 20 microns mesh sieves. DNA of the taeniid eggs retained in the 20 microns sieve was obtained after alkaline lysis and PCR was performed using E. multilocularis species-specific primers. Compared to the parasitological findings after examination of the small intestines of the foxes, the specificity of the PCR was 100% (no false-positive result with 20 foxes free of E. multilocularis) and the sensitivity was 94% (33 positive results from total 35 foxes proven to be infected with E. multilocularis). Both false-negative results were obtained with faeces from foxes harbouring immature worms. Using faecal volumes between 2 and 20 ml, no inhibition of PCR was observed as was demonstrated by the amplification of size-modified target in parallel reactions. The tests were undertaken with fresh faeces stored in 70% ethanol, but egg detection by PCR was also possible after inactivation of eggs by freezing the faeces at -80 degrees C for one week or by incubation at +70 degrees C for 2 h.

Animals↗
PubMed1996

Echinococcus granulosus in Spain: strain differentiation by SDS-PAGE of somatic and excretory/secretory proteins.

A comparison was made, by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), of excretory/secretory (ES)-crude and immunopurified (with the corresponding anti-host serum) hydatid fluids-and somatic (S)-protoscoleces-proteins, from several ovine, equine, swine, bovine and human Echinococcus granulosus Spanish isolates. Likewise, the host influence on parasitic ES protein expression was studied, comparing purified hydatid fluids from ovine and equine cysts obtained from natural hosts and in RNMI mice. Purified hydatid fluids patterns, under reducing conditions, yielded the most precise differentiation of Spanish strains of E. granulosus into three groups (ovine-bovine-human, equine and swine), the finding of a characteristic 82 kDa band in equine isolates, and an unusual arrangement of bands between 50 and 6 kDa in swine samples. In addition, differences were found amongst crude and purified hydatid fluids, especially in bovine and swine isolates. The total protein patterns of protoscoleces were most complex, and therefore could not be used for strain differentiation. Finally, the purified hydatid fluids from cysts developed in natural and experimental hosts showed similar protein patterns, suggesting the lack of host influence, under our experimental conditions, on the expression of parasitic ES proteins.

Animals↗
PubMed1996

Antibody responses against Echinococcus multilocularis antigens in naturally infected Rattus norvegicus.

Two Norway rats, Rattus norvegicus, were found to be naturally infected with Echinococcus multilocularis in Japan. One of them was simultaneously infected with at least three different sized metacestodes of Taenia taeniaeformis. These two R. norvegicus rats and another R. norvegicus naturally infected with T. taeniaeformis and Capillaria hepatica were examined to see if they showed any antibody responses against these two cestode parasites with the view to obtaining more information on the importance of rats as the intermediate host for E. multilocularis. These R. norvegicus showed very poor antibody responses against the two cestode species, although the Wistar rats, R. rattus, experimentally infected with a single smaller sized metacestode of T. taeniaeformis showed stronger responses not only against T. taeniaeformis but also against E. multilocularis. Therefore the three R. norvegicus naturally infected with E. multilocularis and/or T. taeniaeformis demonstrated virtually no immune response, at least against these cestodes.

Animals↗
PubMed1997

Presence of a conserved domain of GATA transcription factors in Echinococcus granulosus.

In order to characterize GATA transcription factors in Echinococcus granulosus, a PCR-based cloning strategy was developed. Degenerate oligonucleotides were designed for the most conserved sequence in GATA proteins that include 20 amino acids of the zinc domain. A 60 bp fragment was isolated that had high homology among this sequence and those reported in other species. An analogous sequence was obtained by performing the same procedure with DNA from the free living platyhelminth Dugesia tigrina. High stringency Southern blotting experiments confirmed the presence of this sequence in the parasite genome.

Amino Acid Sequence↗
PubMed1998

An epidemiological survey of Echinococcus granulosus and other helminths in animal populations in northern Israel.

In a survey carried out during the period May 1995 to November 1996, in communities of various ethnic groups in northern Israel, 206 dogs were examined for Echinococcus granulosus and other intestinal helminth parasites by arecoline hydrobromide purges and the coproantigen-ELISA. The arecoline test was performed close to the owners' homes, using plastic sheets secured to the ground. From 56 dogs examined in the Muslim town of Tamra, six (10.7%) were found to be infected with E. granulosus. Four of them also had a mixed infection of Taenia hydatigena and Dipylidium caninum (two dogs), and the remaining two dogs were infected with either D. caninum or Taenia pisiformis. An additional 18 dogs were infected with either T. pisiformis (eight dogs), D. Caninum (seven dogs), or T. hydatigena (three dogs). Two of these dogs harboured mixed infections whereas the remaining 32 dogs were free of helminths. In the Jewish villages, none of the 150 dogs examined were infected with E. granulosus, although 26 (17.3%) were infected with D. caninum, four (2.7%) with Ancylostoma spp. and one (0.7%) with Toxocara canis. Only one of the 22 stray dogs and none of the 15 jackals examined were infected with E. granulosus. However, 21 (95.4%) of the dogs and 12 (80%) of the jackals harboured helminth infections, including: D. caninum (16 dogs and seven jackals), Ancylostoma spp. (five jackals), T. hydatigena (three dogs), and T. canis (one dog). Approximately 18% of the dogs and 33% of the jackals showed mixed infections with two or more of the above helminths. In the abattoirs, 52 (5.9%) of the 874 sheep and 33 (5.3%) of the 616 goats from 17 herds slaughtered in the Muslim and Druze villages were found to be infected with E. granulosus, compared with a 0% infection rate observed in 93 sheep from two herds in Jewish villages.

Animals↗
PubMed1998

Morphological and developmental characteristics of Echinococcus granulosus derived from sheep, cattle and camels in Iran.

Echinococcus granulosus is widely distributed throughout Iran, where a variety of animals act as intermediate hosts. In this study, the development and morphological characteristics of E. granulosus of sheep, cattle and camel origin were compared and, overall, the characteristics of larval and adult isolates from sheep and cattle, although similar, showed significant differences from those of the camel isolates, especially with respect to the size of rostellar hooks. Differences in the fertility rates of hydatid cyst in sheep, cattle and camel were recorded as 88%, 19% and 70% respectively. In dogs experimentally infected with the three 'strains' of E. granulosus, the rate of development was more rapid in the camel than in the sheep and cattle 'strain'. In addition, adult E. granulosus derived from the camel demonstrate differences in worm length, segmentation and size, shape, number and arrangement of the genitalia when compared with worms derived from sheep and cattle. The epidemiological significance of these findings is discussed in relation to human hydatid infections in Iran.

Animals↗