Echinococcus Granulosus Protoscoleces Research Articles

Preparation and evaluation of fast-dissolving albendazole sulfoxide and praziquantel-loaded polyvinylpyrrolidone nanofiber films by electrospinning

Albendazole sulfoxide (ALBSOX) and Praziquantel (PRZ) are broad-spectrum anthelmintic medications used in the treatment of both humans and animals that are challenging to manufacture for geriatric and pediatric populations and have poor oral bioavailability due to their low water solubility and high lipophilicity. To increase the solubility and bioavailability of the drugs, ALBSOX and PRZ-loaded nanofiber films were prepared using polyvinylpyrrolidone K90 (PVP), a non-toxic, inert polymer with high hydrophilic characteristics. SEM analysis of the produced ALBSOX and PRZ-loaded PVP nanofiber films revealed them to be uniform and smooth, with good uniformity and mechanical properties. The PVP nanofiber films of ALBSOX and PRZ disintegrated in phosphate buffer (pH 6.8) in less than 1 s and increased the solubility of ALBSOX and PRZ about five times compared to pure active ingredients. Furthermore, the prepared ALBSOX and PRZ-loaded PVP nanofiber films were found to have no cytotoxic effect in cell viability studies using L929 cells, and increased the protoscolicidal effect of the ALBSOX and PRZ combination approximately 2-fold in studies of Echinococcus granulosus protoscoleces. The findings of the present study reveal that the developed PVP nanofiber films have the potential to improve the dissolution profile and pharmacological efficacy of the ALBSOX and PRZ.

Evaluation of structural changes of Echinococcus granulosus protoscoleces following exposure to different protoscolicidal solutions evaluated by differential interference contrast microscopy.

The online version contains supplementary material available at 10.1007/s12639-023-01632-4.

Effects of protoscoleces excretory-secretory products of Echinococcus granulosus on hepatocyte growth, function, and glucose metabolism

Cystic echinococcosis (CE) is one of the most widespread and harmful zoonotic parasitic diseases, which most commonly affects the liver. In this study, we characterized multiple changes in mouse hepatocytes following treatment with excretory-secretory products (ESPs) of Echinococcus granulosus protoscoleces (Eg-PSCs) by a factorial experiment. The cell counting kit-8 assay (CCK-8), the 5-ethynyl-2′-deoxyuridine (EdU) assay, and flow cytometry were used to detect the growth of hepatocytes. Inverted microscopy, scanning electron microscopy (SEM), and transmission electron microscopy (TEM) were used to observe the morphology and ultrastructure of hepatocytes. An automatic biochemical analyzer and an ELISA detection kit were used to determine six conventional hepatocyte enzymatic indices, the levels of five hepatocyte-synthesized substances, and the contents of glucose and lactate. Western blot analysis was conducted to analyze the protein expression of three apoptosis-related proteins, Bax, Bcl-2, cleaved caspase-3, and six glucose metabolism pathways rate-limiting enzymes in hepatocytes. The results showed that ESPs inhibited hepatocyte proliferation and promoted hepatocyte apoptosis. The cell membrane and microvilli of hepatocytes changed, and the nucleus, mitochondria and rough endoplasmic reticulum were damaged to varying degrees. The contents of iron, albumin (ALB), uric acid (UA) and urea were increased, and the activities of six enzymes in hepatocytes were increased except for the decrease of transferrin (TRF). The expression levels of all six key enzymes in the glucose metabolism pathway in hepatocytes were reduced. Our characterization provides a basis for further research on the pathogenesis, prevention and treatment of CE.

Report of a two-headed protoscolex of Echinococcus granulosus revealed by phase contrast microscopy in Shiraz, Iran.

Echinococcus granulosus sensu lato can produce cystic echinococcosis (CE)/hydatidosis in different hosts. Each CE cyst can produce numerous protoscoleces by its germinal layer in two forms: evaginated and invaginated. Usually, each protoscolex has one head consisting of a rostellum with two rows of hooks and four suckers. During a study of protoscolicidal agents on sheep Echinococcus granulosus protoscoleces, and investigations on their microscopic changes using Phase contrast microscopy, we observed a case of two- headed evaginated protoscolex. Two heads were attached to a unique bigger size of protoscolex body. Morphological observations showed its dimension around two times of usual protoscolex. There was no space between the bodies, hence one fused body was observed which was clearly shown by Phase contrast microscopy. Each head possessed two rows of hooks. Using micrometry all parts of the two-headed protoscolex, especially hooks were measured and photographed showing all aspects of its morphology including tegument, hooks, and two heads. Specific parts of two-headed protoscolex including suckers, hooks, also calcareous corpuscles were measured. The measurements on the two-headed protoscolex showed that the small hook blade length (SBL) and large hook blade length (LBL) were almost langer than one head protoscolex. A total of 120 calcareous corpuscles in two headed-protoscolex is much higher than one head protoscoleces.

Green Synthesis, Characterization, and Antiparasitic Effects of Gold Nanoparticles against Echinococcus granulosus Protoscoleces.

Echinococcosis, or hydatidosis, is one of the most important zoonotic diseases, which is initiated by the larval stage in the clasts of Echinococcus granulosus. For the treatment of hydatidosis, surgery is still the preferred method and the first line of treatment for symptomatic patients. Unfortunately, most of the scolicidal agents that are injected inside cysts during hydatid cyst surgery have side effects, including leaking out of the cyst and adverse effects on the living tissue of the host, such as necrosis of liver cells, which limits their use. This work was carried out to study the lethal effect of green synthesized gold nanoparticles (Au-NCs) against hydatid cyst protoscoleces. Au-NCs were green synthesized using the Saturja khuzestanica extract. Au-NCs were characterized by UV-visible absorbance assay, electron microscopy analysis, X-ray diffraction (XRD), and Fourier transform infrared (FTIR) spectroscopy. Scolicidal properties of Au-NCs (1-5 mg/mL) were studied against protoscoleces for 10-60 min. The effect of Au-NCs on the expression level of the caspase-3 gene as well as the ultrastructural examination was studied by real-time PCR and scanning electron microscopy (SEM). The cytotoxicity of Au-NCs on hepatocellular carcinoma (HepG2) and normal embryonic kidney (HEK293) cell lines was also studied by the cell viability assay. The obtained Au-NCs are cubes and have an average size of 20-30 nm. The highest scolicidal efficacy was observed at 5 mg/mL with 100% mortality after 20 min of treatment for hydatid cyst protoscoleces. In ex vivo, Au-NCs required more incubation time, indicating more protoscolicidal effects. Au-NCs markedly upregulated the gene level of caspase-3 in protoscoleces; whereas they changed the ultra-structure of protoscoleces by weakening and disintegrating the cell wall, wrinkles, and protrusions due to the formation of blebs. We showed the effective in vitro and ex vivo scolicidal effects of Au-NCs against hydatid cyst protoscoleces by provoking the apoptosis process of caspase-3 activation and changing the ultrastructure of protoscoleces with no significant cytotoxicity against human normal cells. However, additional studies should be conducted to determine the possible harmful side effects and accurate efficacy.

Anti-Tumor Effect of Protoscolex Hydatid Cyst Somatic Antigen on Inhibition Cell Growth of K562.

Today, cancer is one of the most important causes of death in the world, and so far, many treatment methods have been used in this field. Immunotherapy is considered one of the newest developments in this science, and it is still being investigated in some forms in different cancers and with a variety of antigens as well. One of the subsets of cancer immunotherapy is its treatment using parasitic antigens. The present study evaluated the effect of using somatic antigens of protoscoleces of Echinococcus granulosus on K562 cancer cells. In this study, hydatid cysts' protoscolex antigens were extracted, purified, and added to K562 cancer cells at three concentrations (0.1, 1, and 2mg/ml) and on three times (24, 48, and 72h). The number of apoptotic cells was compared to the control flask. The antigen concentration of 2mg/ml was used as a control sample to investigate its cytotoxic effect on the growth of healthy HFF3 cells. Annexin V and PI tests were also performed to differentiate apoptosis from necrosis. In flasks treated with hydatid cyst protoscolex antigen, all three concentrations significantly reduced the growth of cancer cells compared with the control flask, and concentration 2 of crude antigen significantly caused the death of cancer cells. Furthermore, more cancer cells underwent apoptosis by increasing the time of exposure to the antigen. On the other hand, flow cytometry results also showed that the amount of apoptosis has increased compared to the control group. In fact, Protoscolex hydatid cyst somatic antigens induce programmed cell death in K562 cancer cells while not having a cytotoxic effect on normal cells. Therefore, it is suggested to do more research on the anti-cancer and therapeutic properties of the antigens of this parasite.

Effect of Some Plants Extracts on the Viability of Protoscoleces in Vitro

This study was conducted to evaluate the protoscolicidal effect of four plants alcoholic extracts on protoscoleces of the parasite Echinococcus granulosus in vitro. The plants that had been used in this study were: Viscum album (Mistletoe) and Prunus armeniaca (Apricot bitter kernels) with concentrations of 375, 750 and 1500 mg/ml, also Syzygium aromaticum (Clove flower buds) and Capsicum frutescence (Chili pepper) with concentrations of 375, 750 and 1500 µg/ml. All extracts were treated at four periods (15, 30, 45 and 60 min.).
 In vitro, the high percentage of protoscoleces killing was observed in V. album alcoholic extract (80.7%) in the high concentration 1500 mg/ml at 60 min., and the lowest percentage of protoscoleces killing (62%) was in the concentration 375 mg/ml at 15 min.
 
 armeniaca alcoholic extract high percentage of killing (70.2%) was also at the high concentration 1500 mg/ml at 60 min., and the lowest percentage of protoscoleces killing (50.4 %) at the concentration 375 mg/ml at 15 min.
 
 The high percentage of protoscoleces killing of alcoholic extract of S. aromaticum (73.7%) was at the concentration 1500 µg/ml at 60 min. and the lowest percentage (58.7%) was at the concentration 375 µg/ml at 15 min.
 
 frutescence alcoholic extract high percentage of protoscoleces killing (71.4 %) was at the concentration 1500 µg/ml at 60 min. and the lowest percentage (40.6 %) was at the concentration 375 µg/ml at 15 min.

Evaluation of scolicidal potential of salicylate coated zinc nanoparticles against Echinococcus granulosus protoscoleces

Echinococcosis is a zoonotic disease caused by larval stages of the Echinococcus genus (metastasis). In this study, salicylate-coated Zinc oxide nanoparticles (SA-ZnO-NPs) were fabricated and characterized by SEM, FTIR and XRD analytical techniques. After that, different doses of SA-ZnO-NPs, SA and ZnO-NPs were taken to assess scolicidal potency. Scanning electron microscopy (SEM) micrographs were also used to evaluate the morphological deformities of treated protoscoleces. Furthermore, Caspase-3&7 inductions were examined in protoscoleces cysts treated with all formulations. Based on SEM and DLS analyses, the size of SA-ZnO-NPs was between 30 and 40 nm, with a spherical shape. The FTIR spectrum verified the presence of SA functional groups on the ZnO coating. At 20 min, SA-ZnO-NPs at 2000 μg/ml exhibited the greatest activity on protoscolices with 100% mortality, followed by ZnO-NPs at 1500 μg/ml at 10 min and SA alone at 2000 μg/ml at 30 min. The activation of Caspase-3&7 apoptotic enzyme was determined for 2000 μg/ml of SA-ZnO-NPs, ZnO-NPs and SA to be 16.4, 31.4, and 35.7%, respectively. The SEM image revealed apoptogenic alterations and the induction of tegument surface wrinkles, as well as abnormalities in rostellum protoscolices. According to the current study, SA-ZnO-NPs have a high mortality rate against hydatid cyst protoscolices. As a result, further studies on the qualitative assessment of these nanoformulations in vivo and preclinical animal trials seem to be required. Furthermore, the adoption of nano-drugs potentially offers alternative therapeutic approaches to combat hydatid cysts.

Effect of selenium nanoparticles against protoscoleces of Echinococcus granulosus in vitro and hydatid cysts in mice

This study determined the influence of selenium nanoparticles on the vitality of the protoscoleces of Echinococcus granulosus in vitro; seven concentrations were used: 50, 100, 150, 200, 250, 350, and 500 µg/ml for different exposure times: ten, twenty, thirty, and sixty minutes, respectively. Albino mice Mus musculus were injected with protoscoleces exposed to nanoparticles at 100, 150, and 200 µg/ml concentrations for 60 minutes. In contrast, control groups were injected with non-exposed protoscoleces. Mice were dissected three, four-, and five-months post infestation. Many criteria were relied on: numbers, weights, diameters of growing hydatid cysts, and their reduction proportion. The outcomes uncovered an apparent influence of selenium nanoparticles on the viability of protoscoleces of Echinococcus granlosus by the increase in exposure time in vitro, as well as diminish in the numbers of the larvae in processed mice versus the unprocessed collection; no cysts evolved inside processed mice at the concentration 200 µg/ml, three- and four-months post-infection. In contrast, the reduction rate was 90% in mice injected with exposed protoscoleces at the concentration of 150 µg/ml next 4 and 5 months of infection.

Application of microwave against Protoscoleces of Echinococcus granulosus in vitro and Secondary Hydatid Disease in BALB/c Mice

Application of microwave against Protoscoleces of Echinococcus granulosus in vitro and Secondary Hydatid Disease in BALB/c Mice

Anti-helminthic effects and cellular mechanisms of Astragalus ecbatanus extract against Echinococcus granulosus protoscoleces.

We aimed to investigate the in vitro and ex vivo anti-helminthic effects of Astragalus ecbatanus chloroform extract (AECE) as well as its cellular mechanisms against Echinococcus granulosus protoscoleces. In vitro and ex vivo anti-helminthic effects of AECE on E. granulosus protoscoleces were evaluated through eosin exclusion test. Effects of AECE on induction of the caspase-3 like activity, the permeability of plasma membrane were also determined. The effects of AECE against the human hepatocyte carcinoma (HepG2) and human embryonic kidney (HEK293T) cells were determined by MTT ([3-(4.5-dimethylthiazol-2-yl)-2.5-diphenyl tetrazolium bromide)]) test. The findings revealed that AECE especially at 45mg/mL displayed potent in vitro and ex vivo anti-helminthic effects against E. granulosus protoscoleces (p < 0.001). After treatment of protoscoleces with AECE, the caspase-3 enzyme activity and the rate of permeability of plasma membrane was dose dependently (P < 0.001) increased. The 50% cytotoxic concentration (CC50) value of AECE for normal and cancer cells lines 4.62mg/mL and 1.89mg/mL, respectively. The findings of the current study revealed the favorable in vitro and ex vivo anti-helminthic effects of AECE against protoscoleces of E. granulosus. However, further surveys in animal model are required to elucidate different aspects of this extract before use in clinical phases.

2-Deoxy-D-glucose and combined 2-Deoxy-D-glucose/albendazole exhibit therapeutic efficacy against Echinococcus granulosus protoscoleces and experimental alveolar echinococcosis.

2-Deoxy-D-glucose (2-DG) is a glucose analog used as a promising anticancer agent. It exerts its effects by inhibiting the glycolytic energy metabolism to deplete cells of energy. The larval stage of Echinococcus relies on glycolysis for energy production. Therefore, in this study, we investigated the in vitro and in vivo efficacy of 2-DG against the larval stage of Echinococcus granulosus and E. multilocularis. 2-DG exhibited significant time- and dose-dependent effects against in vitro cultured E. granulosus protoscoleces and E. multilocularis metacestodes. A daily oral administration of 500 mg/kg 2-DG in E. multilocularis-infected mice effectively reduced the weight of metacestodes. Notably, the combination treatment, either 2-DG (500 mg/kg/day) + albendazole (ABZ) (200 mg/kg/day) or 2-DG (500 mg/kg/day) + half-dose of ABZ (100 mg/kg/day), exhibited a potent therapeutic effect against E. multilocularis, significantly promoting the reduction of metacestodes weight compared with the administration of 2-DG or ABZ alone. Furthermore, the combination significantly promoted apoptosis of the cells of metacestodes and inhibited glycolysis in metacestodes, compared with the administration of 2-DG or ABZ alone. In conclusion, 2-DG exerts an effective activity against the larval stage of Echinococcus. Thus, it may be a promising anti-Echinococcus drug, and its combination with ABZ may provide a new strategy for the treatment of echinococcosis in humans.

In vitro, effects of Praziquantel and alcoholic and aqueous crude extracts of Citrullus colocynthis and Eucalyptus globulus on the viability of protoscoleces of Echinococcus granulosus

Iraqi flora is rich in plants and the possibility of finding new antimicrobial agents still widely ahead, therefore, the present work evaluated the effects of alcoholic and aqueous crude extracts of Citrullus colocynthis; Eucalyptus globulus leaves and Praziquantel drug (as scolicidal agents) on the viability of Echinococcus granulosus protoscoleces in vitro. Live protoscoleces obtained from the liver of naturally infected sheep. The viability of protoscoleces was determined by their motility and flame cells activity and staining method using 0.1% eosin solution. The number of viable protoscoleces was counted and distributed into test tubes (4 tubes for each treatment of different concentrations) and 2 tubes were kept as control without treatments. The percentage viability of protoscoleces was ranged between 93.7 to 95.3. Praziquantel drug reduced the survival of protoscoleces during seven sequenced days post incubation. Drug effects showed more influence in both concentrations of 450 μg/ml from day 4 and 600 μg/ml from day 3 post incubation, respectively. The methanolic extract of C. colocynthis reduced the viability of protoscoleces during eight sequenced days post incubation from 96% to zero%. The effects were marked highly in the concentrations of 300 μg/ml from day 3 post incubation. Aqueous fruit extracted decreased in the viability of protoscoleces. In the minimum used concentration (75μg/ml), the viability of protoscoleces reduced from 96% to reach zero% at day eight post incubation, whereas in the maximum concentration used (300 μg/ml), the viability reached to zero% after four days. All concentrations of leaves methanolic and aqueous extracts of E. globulus have an antiparasitic activity against the protoscoleces viability, but the greater effect was to 600 μg/ml that kill all protoscoleces in 6 days after treatment.

Determination of extracellular traps structures from sheep polymorphonuclear leukocytes to Echinococcus granulosus protoscoleces

It was aimed to detect extracellular traps structures from sheep polymorphonuclear leukocytes (PMN) after being confronted with Echinococcus granulosus protoscoleces in vitro. Also, the effect of cyst fluid was examined on the development of extracellular traps. At the end of the incubation for 1 h, the extracellular traps augmented with neutrophil elastase, histone (H3) and myeloperoxidase were visualized in the protoscoleces-PMN co-culture microscopically. Some protoscoleces lysed and the chitinous hooks released were surrounded by the extracellular traps. The other protoscoleces were still intact and the extracellular trap structures were observed around them. The relationship between the extracellular DNA contents and the protoscoleces concentration was not found statistically significant (P > 0.05). The extracellular DNA amount in the co-cultures diluted in RPMI-1640 increased with the incubation time (P < 0.05). However, the time-dependent relationship was not found in the co-cultures diluted in the cyst fluid (P > 0.05). The difference in the extracellular DNA amount was detected as statistically significant (P < 0.05) between the two co-culture groups (diluted in RPMI-1640 or the cyst fluid), except for 30 min incubation. To the Author's knowledge, NETosis reaction was firstly observed in sheep PMN after being confronted with protoscoleces in vitro. The cyst fluid had some negative effects on the development of extracellular traps from sheep PMNs at the 1-h incubation time. It should be investigated which molecules are responsible for NETosis inhibition in hydatid cyst fluid. Future studies may clarify whether neutrophils fight with protoscoleces by using their different mechanisms.

Electrical potentials of protoscoleces of the cestode Echinococcus granulosus from bovine origin

Larval stages of taeniid Echinococcus granulosus are the infective forms of cystic echinococcosis or hydatidosis, a worldwide zoonosis. The protoscolex that develops into the adult form in the definitive host is enveloped by a complex cellular syncytial tegument, where all metabolic interchange takes place. Little information is available as to the electrical activity of the parasite in this developmental stage. To gain insight into the electrical activity of the parasite at the larval stage, we conducted microelectrode impalements of bovine lung protoscoleces (PSCs) of Echinococcus granulosus in standard saline solution. We observed two distinct intra-parasitic potentials, a transient peak potential, and a stable second potential, most likely representing tegumental and intra-parasitic extracellular space electrical potential differences. These values changed on the developmental status of the parasite, its anatomical regions, or time course after harvesting. Changes in electrical potential differences of the parasite provide an accessible and valuable parameter for the study of transport mechanisms and potential targets for developing novel antiparasitic therapeutics.

In vitro protoscolicidal effects of lithocholic acid on protoscoleces of Echinococcus granulosus and its mechanism

Surgery has been found to be the best choice of treatment for hydatidosis. However, leakage of cyst contents during surgery is the foremost reason for recurrence of hydatidosis. In this study, we investigated the in vitro efficacy of lithocholic acid (LCA) against Echinococcus granulosus protoscoleces. The protoscoleces were divided into a control group, an albendazole (ABZ) positive control group and LCA intervention groups at concentrations of 0.5, 1, 2, and 3 mmol/L and stained with 0.1% eosin for observation using an inverted microscope; the protoscolecal ultrastructure was examined with SEM and TEM; the activities of ROS, SOD, and caspase-3 were investigated using an ROS kit, SOD kit, and caspase-3 kit, respectively; the contents of HO-1 and NQO-1 were analyzed by enzyme-linked immunosorbent assay; and the expression level of cytochrome c (Ctyc) was analyzed by western blotting. Results: As the concentration of LCA increased, the survival rate of protoscoleces gradually decreased. The microstructure shows that the external shape and internal structure were gradually deformed and collapse. SOD, GSH, HO-1 and NQO-1 decreased more significantly in the 3 mmol/L LCA group. However, ROS levels gradually increased. LCA treatment for 3 days at all concentrations significantly increased caspase-3 activity and expression in a dose-dependent manner. LCA decreased the level of Ctyc protein in vitro. LCA demonstrated a parasiticidal effect on the protoscoleces of Echinococcus granulosus in vitro. LCA may induce apoptosis of E. granulosus protoscoleces by oxidative stress and mitochondrial pathways.

Expression profiling of exosomal miRNAs derived from different stages of infection in mice infected with Echinococcus granulosus protoscoleces using high-throughput sequencing.

Echinococcosis is a worldwide zoonosis. The mechanism of the establishment, growth, and persistence of parasites in the host has not been fully understood. Exosomes are found to be a way of information exchange between parasites and hosts. They exist in various body fluids widely. There are few studies on host-derived exosomes and their miRNA expression profiles at different infection time points. In this study, BALB/c mice were intraperitoneally infected with protricercariae. Exosomes were extracted from plasma (0, 3, 9, and 20weeks post infection), and the expression profiles of exosome miRNA in the peripheral blood of mice were determined using RNA-sequencing. Compared to the 0week groups, 24, 35, and 22 differentially expressed miRNAs were detected in infected mouse at the three infection stages, respectively. The results showed that there were significant differences in the miRNAs of exosomes at different infection time points. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis were used to annotate the different miRNAs. The results showed that the biological pathways of parasites changed significantly at different stages of infection, with many significant and abundant pathways involved in cell differentiation, inflammation, and immune response, such as MAPK signaling pathway, Th17 cell differentiation, Wnt signaling pathway, FoxO signaling pathway, Notch signaling pathway, etc. These results suggest that miRNA may be an important regulator of interactions between Echinococcus granulosus and host. The data provided here provide valuable information to increase understanding of the regulatory function of microRNAs in the host microenvironment and the mechanism of host-parasite interaction. This may help us to find targets for Echinococcus granulosus to escape host immune attack and control Echinococcus granulosus infection in the future.

Antiparasitic effects of Elettaria cardamomum L. essential oil and its main compounds, 1-8 Cineole alone and in combination with albendazole against Echinococcus granulosus protoscoleces

BackgroundThe present investigation aims to determine the chemical structure and protoscolicidal effects of Elettaria cardamomum L. essential oil (ECEO) and its main compounds 1–8 cineole alone and along with albendazole (ALZ) against Echinococcus granulosus protoscoleces in vitro and ex vivo. We also decided to evaluate some cellular mechanisms such as the apoptotic activity and the permeability of plasma membrane of protoscoleces treated with ECEO and 1–8 cineole. MethodsHydatid cyst protoscoleces were divided into seven groups including protoscoleces treated with ECEO 50 µl/mL (T1), protoscoleces treated with ECEO 100 µl/mL (T2), protoscoleces treated with ECEO 200 µl/mL (T3), protoscoleces treated with 1–8 cineole 100 µg/mL (T4), protoscoleces treated with 1–8 cineole 200 µg/mL (T5), protoscoleces treated with 1–8 cineole 100 µg/mL + albendazole 50 µg/mL (T6), and protoscoleces treated with 1–8 cineole 200 µg/mL + albendazole ALZ-50 µg/mL (T7). The viability of protoscoleces were recorded by eosin staining examination. Moreover, the induction of apoptosis and the plasma membrane permeability of the protoscoleces treated with ECEO and 1–8 cineole were evaluated. ResultsThe highest protoscolicidal effect of ECEO was observed at the dose of 200 µl/ml (T3). 1,8-Cineole alone and combined with ALZ, particularly at the dose of 200 µg/ml (T5 and T7), destroyed the 100% protoscolices after 10 min incubation. The ECEO (T1-T3) and 1–8 cineole alone (T4 and T5) and in combination with ALZ (T6 and T7) took longer to display their protoscolicidal effect ex vivo. The obtained results of relative fuorescent items exhibited that the protoscoleces incubated with ECEO and 1,8-Cineole, alter the permeability of plasma membrane by Sytox Green with increasing the concentration. The findings revealed exhibited that ECEO and 1,8-Cineole increasingly and dose-dependently induced activation of caspase-3 enzyme ranging from 6.8 to 23.3%. ConclusionOur obtained results revealed that ECEO and its main compound, 1,8-Cineole exhibited the potent protoscolicidal in vitro and ex vivo; and if more research is done on their efficacy and toxicity in animal models and even clinical setting, it can be suggested as a protoscolicidal agent to use during hydatid cyst surgery.

Effect of ultrasound on protoscoleces of Echinococcus granulosus in vitro and in vivo

The current study was designed to investigate the effect of ultrasound on the protoscoleces of Echinococcus granulosus by enforcing steady numeral of frequencies during a certain interval (20000 pulse/s) (1.8 w/cm2), using exposure time 30,20,15,12,10 and 5s, individually and respectively. Consequently, six albino mice groups were immunized against cystic echinococcosis, which injected with exposed protoscoleces, to acquire specific cell-mediated immunity, called delayed type-hypersensitivity (DTH) which assessment by measurement the foot pad density. The results displayed significant excess (P≤ 0.001) of DTH by increase of foot pad thickness in injected groups. The results showed maximum thickness of 1.54 mm, 1.4mm, 0.9 mm. after 3h, 24h, and 48h post - injection, respectively, that compared with the thickness of control group 1.072, 0.638, 0.328 mm, respectively, during five months of experiment. The present research exhibited the action of ultrasonication technique on the viability of Echinococcus granulosus protoscoleces in vitro. Conclusion, Ultrasound frequencies used in the present study could be have consequential impact on the cellular immunity in albino mice.

Genome-wide transcriptome analysis of the early developmental stages of Echinococcus granulosus protoscoleces reveals extensive alternative splicing events in the spliceosome pathway

BackgroundThe complex life cycle of Echinococcus granulosus involves numerous environmental conditions within different intermediate and definitive hosts. This requires adaptation at different levels of transcript regulation. Alternative splicing (AS) and the related cellular functions as one of the major fields of post-genomics has been poorly studied in tapeworms. In the present study, we investigated AS events and their potential biological effects in E. granulosus.MethodsWhole transcriptome sequencing data of four groups of protoscoleces were prepared for RNA-seq library construction. Fresh protoscoleces were either used as non-induced controls (NT group) or incubated for 15 min with pepsin (PEP group) and cultivated in a biphasic medium for 12 and 24 h (12 and 24 h groups). The frequency and different types of AS events were identified using rMATS software. Functional annotations and gene ontology of differential AS (DAS) genes were performed using Blast2GO software. AS events were experimentally validated by PCR on the protoscolex cDNAs using specific primers for each gene.ResultsAt least one AS event was found in 38.1% of the genes (3904 out of 10,245) in the protoscoleces during early strobilar development. The genes were associated primarily with cellular and metabolic processes and binding and catalytic activity. KEGG pathway analysis of DAS events revealed a number of genes belonging to different components of the spliceosome complex. These genes tended to belong to common SR proteins, U1-related factors, U2-related factors, complex A-specific factors and other splicing-related proteins.ConclusionsThe high number of AS events in the transcriptome regulatory mechanisms indicates the essential rapid molecular processes required by the parasite for adaptation in different environments.Graphical