Analysis of Protoscoleces-specific Antigens from Echinococcus Granulosus with Proteomics Combined with Western Blot

Objective To establish and optimize the proteomic analysis of protoscoleces-specific antigens from Echinococcus granulosus. To provide a foundation for identifying specific antigens in the soluble proteins of E. granulosus protoscoleces for further research. Methods Brood capsules were collected aseptically from fertile E. granulosus cysts from the livers of an infected patient. The fertile E. granulosus cysts were fractured, and protoscoleces were collected by centrifugation. The soluble proteins of protoscoleces were acquired using the 2D Quant kit according to the manufacturer＇s instructions. We employed two-dimensional electrophoresis （2-DE） combined with immunoblot assay （Western blot） to analyze the soluble components of E. granulosus protoscoleces antigens. The 2-DE and immunoblot maps obtained were analyzed with PDQuest 8.0 image analysis software. Results About 233 soluble protein spots were identified with Coomassie-stained gels. Most of the proteins had a molecular weight of 16 000 Da to 117 000 Da, and an isoelectric point value of 3.0 to 10.0. 2-DE immunoblot was conducted and 57 specific antigen spots were observed, among which 23 spots were identified. Conclusion 2-DE combined with Western blot is the key to successful proteomic analysis and presents a new possibility for searching the specific E. granulosus protoscoleces antigens.

<i>In Vivo</i>and <i>In Vitro</i>Survival Rates of Protoscoleces Kept at Different Constant Temperature

An experiment was conducted on the survival rate of protoscoleces derived from hydatid cyst brought from infected camels in Tambool area (North Eastern Sudan). The survival rate was studied in vivo and in vitro through exposure to different constant temperatures to measure their viability. The results revealed that the protoscoleces remained viable in vitro for 20 days at 4°C, 7 and 5 days at 25°C and 37°C, respectively, while at 45°C the protoscoleces were immediately dead. This means that the infection rate of hydatid cyst becomes high in relatively cold seasons (winter and autumn). In the in vivo study, each mouse (five groups, four mice in each) was inoculated intraperitoneally with 2000 protoscoleces exposed to the same different constant temperature, as previously used. 6 months later, the mice in different groups were sacrificed and necropsied to study the development of hydatid cyst inside. The results showed that the rate of development inside the mice was clear in the first and the second groups (4°C and 25°C), however, in the other 2 groups (37°C and 45°C), no sign of development of the cyst was observed, with the presence of few number of hooks compared to the control group, which showed significant difference in the development of hydatid cyst.

Experiment of Hydatid Cyst in Two Strains (Camels and Goats) in Saudi Arabia

Hydatid cyst disease is one of the most parasitic diseases transmitted from carnivores animals (such as dogs) to humans and causes a deterioration in the general health. Its transmission to herbivores animals led to a substantial economic loss in the meat productivity and reduced its quality. The increase of acquisition of dogs at homes nowadays led to an increase in the risk of infection with many parasitic diseases such as hydatid cyst disease. The present study was conducted to provide a modern view on the current status of hydatid cyst disease among slaughtered animals in Jeddah through periodic visiting to a slaughter house in North Jeddah. The mice prepared have been infected in the peritoneal membrane with 0.2 ml of hydatid cyst fluid containing ±2000 protoscoleces from fertile cysts obtained from an infected goat’s liver which resulted in a 75% infection rate. Mice infected with hydatid fluid from fertile cysts obtained from an infected camel’s lungs showed a result of 11.1% infection rate. The cysts appeared in the peritoneal and abdominal cavity.

Anti-parasitic effects of water-soluble alkaloid fractions from ethanolic extracts of Sophora moorcroftiana seeds in Caenorhabditis elegans

Parasite infections of humans and animals remain a major global health problem, with limited choice of drugs being available to the treatment of parasitosis in the clinic. Sophora moorcroftiana(S. moorcroftiana) is a shrub that grows in Tibet Plateau of China. Decoction of the seeds has been used as a traditional Tibetan medicine to treat parasitosis for years. But the anti-parasitic effects of water-soluble fractions in the seeds need further investigation. In the present study, the water-soluble alkaloid fractions(E2) were obtained from S. moorcroftiana seeds by refluxing extraction with 60% ethanol and low polarity fraction(E2-a) and high polarity fraction(E2-b) were subsequently isolated from E2 using column chromatography. As a parasite model, Caenorhabditis elegans(C. elegans) were treated with different fractions and their survivals were recorded. The results showed that that E2-a induced a lower survival rate in C. elegans than E2-b and E2. The protoscoleces of Echinococcus granulosus(E. granulosus) were cultured in the presence of E2-a. Compared with E2-b and E2, protoscoleces exhibited decreased survival rate following E2-a treatment. Furtherly, the effects of E2-a on the behavior, brood size, and lifespan of the worms were investigated. Body bend frequencies of the worms treated with the high concentration of E2-a were reduced by two-thirds compared with the control group(P < 0.01). Compared with non-E2-a-treated group, exposure of nematodes to E2-a led to a decrease in head thrashes and pharyngeal pumps frequency(P < 0.01). E2-a treatment resulted in a significantly lower brood size(P < 0.01). Additional E2-a treatment induced a significantly shortened lifespan, compared with the control(P < 0.05). These findings indicated that water-soluble fraction E2-a from S. moorcroftiana seeds was a potential helminthic agent.