Hydatid Cyst Protoscoleces Research Articles

Effect of Temperature and Salinity on Survival of Protoscoleces of Hydatid Cyst in Liver In Vitro.

Hydatid cyst is the metacestode stage of Echinococcus granulosus that occurs in herbivores and humans as intermediate hosts by consuming parasite eggs through forage and vegetables. Carnivores, as definitive hosts, become infected by consuming infected vesicles of herbivores. The most effective treatment for a hydatid cyst is surgical operation. Inactivating E. granulosus protoscoleces through heating, cooling, or chemicals such as sodium chloride can be considered an effective method for controlling hydatidosis in both humans and animals. The main objective of this study was to evaluate the effect of different temperatures and salinity conditions on the survival of Echinococcus granulosus protoscoleces. For this purpose, 50 g of infected liver (in triplicate) was separately treated with different temperatures (+10°C, +50°C, +60°C, +72°C, and -20°C) and concentrations of sodium chloride (5%, 10%, 15%, and 20%) for 3, 6, 12, 24, 48, and 72 h. Additionally, 50 g of infected liver was stored separately in the refrigerator (+4°C) as a control group. The survival rate of the protoscoleces was evaluated by staining with 1% eosin under a light microscope. The results showed that the protoscoleces were significantly affected, with 100% mortality at -20°C after 0.5 h, and complete death at +72°C, +60°C, +50°C, and +10°C after 1, 1.5, 3, and 24 h, respectively (p < 0.005). Similarly, the protoscoleces in the liver mass survived at 5% NaCl after 3 h but died at 10% after 24 h, at 15% after 12 h, and at 20% after 6 h. It is concluded that exposing the liver infected with protoscoleces hydatid cyst to a temperature of -20°C and a sodium chloride concentration of 10% for 24 h is suitable for inactivating the protoscoleces.

Synthesis of Silver Nanoparticles Using Haplophyllum robustum Bge. Extract: Antibacterial, Antifungal, and Scolicidal activity against Echinococcus granulosus Protoscolices

Background: Silver nanoparticles (AgNPs) biosynthesized via the deployment of plant extractives have garnered much attention, especially due to their antimicrobial properties. Herein, the green synthesis of silver nanoparticles has been accomplished using the aqueous extract of Haplophyllum robustum, which includes a study of its antibacterial, antifungal, and scolicidal activity. Methods: The preparative process was followed by characterization using UV-Vis spectroscopy, and the ensuing spherical AgNPs of average size 7-25 nm were identified by Dynamic Light Scattering (DLS), X-Ray Diffraction (XRD), Fourier Transform Infrared Spectroscopy (FTIR), Scanning Electron Microscopy (SEM), and Transmission Electron Microscopy (TEM). The antibacterial, antifungal, and scolicidal activities of AgNPs were assessed by deploying disc diffusion and microdilution methods against four standard bacteria and four typical Candida species and liver hydatid cyst protoscoleces, where they exhibited good biological activity. Results: The results showed that the greener synthesis of silver nanoparticles using the aqueous extract of renewable and abundant H. robustum plant is a simple, inexpensive, and safer alternative that does not use any toxic or harmful substances. Conclusion: Thus, with minimal or no side effects, this approach to AgNPs bodes well for their appliances as antibacterial, antifungal, and scolicidal agents.

High potency of linalool-zinc oxide nanocomposite as a new agent for cystic echinococcosis treatment.

This current in vitro, ex vivo, and in vivo research aims to evaluate and analyze the linalool-zinc oxide nanocomposite (Lin-ZNP) for treating cystic echinococcosis. Lin-ZNP was synthesized using an ethanolic solution of polyvinyl alcohol. The protoscolicidal effects of Lin-ZNP were tested on hydatid cyst protoscoleces (PTS) in both in vitro and ex vivo by eosin exclusion test. The study also examined the impact on caspase-3 gene expression and the external structure of PTS. The in vivo effect was measured by examining hydatid cysts' quantity, dimensions, and weight in mice intraperitoneally infected with 0.5 mL of PTS solution containing 1,000 PTS. The antioxidant and inflammatory cytokine gene expression levels were examined using real-time PCR. Lin-ZNP significantly (P < 0.001) killed the PTS in both in vitro and ex vivo in a dose- and time-dependent manner. The treated PTS exhibited creases and protrusions as a result of bleb formation and upregulation in the gene expression of caspase-3. Upon treatment with Lin-ZNP, there was a significant (P < 0.001) reduction in the number, diameter, and weight of the hydatid cysts. Treatment with Lin-ZNP nanocomposite led to a significant increase in the expression of antioxidant genes and a notable decrease in oxidative stress markers, and the expression levels of IL-4 and IL-10. Lin-ZNP has the potential to act as a scolicidal agent and demonstrates promise in controlling hydatid cysts in a mouse model, attributed to its antioxidant and anti-inflammatory properties. However, additional studies in clinical trials are needed to confirm the use of Lin-ZNP for treating hydatidosis.

Human and camel cystic echinococcosis – a polyclonal antibody-based sandwich ELISA for its serodiagnosis with molecular identification

Cystic echinococcosis (CE) is an emergent neglected disease affecting human and animals in Egypt with a wide distribution and incidence. This study aimed to evaluate the use of a polyclonal antibody-based sandwich ELISA in the detection of Echinococcus granulosus antigen in human and camel sera. Hydatid cyst protoscoleces antigen (PsAg) was isolated from hydatid cysts collected from naturally infected camel livers and lungs. PsAg was used for immunization of rabbits to raise IgG polyclonal antibodies (IgG PsAb). IgG PsAb were then precipitated, purified using Protein-A Sepharose gel and labeled with horseradish peroxidase enzyme. We assayed the purity of the IgG PsAb, and the two prepared E. granulosus antigens CPsAg from camel cysts and HPsAg from human cysts by Sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The resulted protein bands of the prepared CPsAg appeared at different molecular weights: 180, 90, 68, 54, 42 and 22 kDa while, HPsAg shared with it in 4 common bands at 68, 54, 42, and 22 kDa. The purified IgG PsAb had been resolved at two bands at 52 kDa and at 32 kDa. Sandwich ELISA were performed for the detection of circulating E. granulosus antigens in sera of human (n = 183) and camels (n = 190). The purified IgG PsAb showed strong reactivity against E. granulosus infected human and camel samples and no cross reactivity neither with free-healthy negative sera nor with others parasitic diseases (Schistosomiasis, Fascioliasis, Toxoplasmosis, Ancylostomiasis for human samples and Fascioliasis, ticks’ infestation, Eimeriosis, Cryptosporidiosis, Nasal myiasis, Toxoplasmosis for camel samples). The sensitivity of the assay was 98.25% (56/57) and 96.9% (31/32) against human and camel samples, respectively. Specificity was 100% in both human and camel samples. Sandwich ELISA detected CE in 33.3% (24/72) and 55.6% (50/90) random human and camel samples, respectively. Indirect ELISA, using CPsAg, was used for detection of antibodies in positive human and camels’ sera and detected 96.5% (55/57) and 93.8% (30/32) of human and camel samples, respectively. In our study, Genomic DNA was extracted from protoscoleces fluid of human liver hydatid cysts to identify the Echinococcus sp. isolate based on NADH dehydrogenase subunit 1 (NAD1) gene by Polymerase Chain Reaction (PCR) and the isolate (GenBank: OP785689.1) were identified as E. granulosus sensu lato genotype. In conclusion, Sandwich ELISA technique was found to be a potent and sensitive assay for detection of hydatid antigen in both human and camel samples.

Larvicidal effect of zirconium oxide nanoparticles in protoscoleces of hydatid cysts in the laboratory and mice

Larvicidal effect of zirconium oxide nanoparticles in protoscoleces of hydatid cysts in the laboratory and mice

Zirconium oxide nanoparticles: Protocolicidal effect against hydatid cyst protoscoleces with interleukins evaluation in mice

Zirconium oxide nanoparticles: Protocolicidal effect against hydatid cyst protoscoleces with interleukins evaluation in mice

The co-infection of pulmonary hydatid cyst, lophomoniasis and tuberculosis in a patient with resistant respiratory symptoms; a case report study

BackgroundLophomonas blattarum is a rare protozoan that causes opportunistic infections, and the co-infection of lophomonas with tuberculosis and human hydatidosis is a serious public problem in the co-endemic areas of developing countries.Case reportWe presented a 58-year-old female with fever, losing weight, and cough with whitish-yellow sputum that started one month ago. Increasing inflammatory markers and hypereosinophilia in laboratory tests, and a cavity with thick, regular walls and undulating air-fluid levels measuring 43 × 30, evident in the upper segment of the right lower lobe (RLL), along with consolidation and the ground glass opacity of the upper segment and posterior basal of the RLL is apparent in CT scan were reported. Then, a bronchoscopy was requested, and the BAL specimen reported a negative fungal and bacterial infection in the samples. Several live and oval flagellated lophomonas protozoa, hydatid cyst protoscoleces (the larval forms of the parasites), and M. tuberculosis were observed in microscopic evaluation. The patient was treated with metronidazole, oral albendazole, and a combination of TB regimen.ConclusionPhysicians should always consider the possibility of co-infections of lophomonas with tuberculosis and human hydatidosis and investigate patients with risk factors such as immunodeficiency conditions or treated with immunosuppressive medications.

Toxicity of geranium oil, geraniol and their nanoemulsions on protoscoleces of hydatid cyst under in vitro conditions

Surgery is the main efficacious treatment for many cases of cystic echinococcosis by removing Echinococcus granulosus cysts. However, to reduce risk of cyst spillage and insemination of the content, using a scolicidal agent is crucial. Considering side effects of available scolicidals, and growing nano-technological approaches in novel pharmaceuticals, the present study aimed to find out the scolicidal activity of geraniol (GL), geranium oil (GM), and their developed nanoemulsions (Nano-GL and Nano-GM) on the protoscoleces of E. granulosus. Nanoemulsions were developed by ultrasonication emulsification and characterised by dynamic light scattering method (DLS). The scolicidal effect of GL, GM, Nano-GL, and Nano-GM at different concentrations of 1, 2.5, 5, 10, 25, and 50 µg/mL were determined after 0.5, 1, and 2 hours of incubation. Mortality rates were measured by eosin exclusion test. The average droplet size for Nano-GM and Nano-GL were 124.8 and 88.59 nm, respectively. After one hour of exposure, all tested concentrations of GL and Nano-GL resulted in higher than 90% mortality rates, while GM and Nano-GM killed 70.66% and 90.33% of protoscoleces, respectively. Based on 50% lethal concentration, Nano-GL was significantly more potent than Nano-GM (LC50: 4.52 vs 102.95 μg/mL). Results of scanning electron microscopy revealed tegumental disruption in the treated protoscoleces. This study described an easily applicable and eco-friendly procedure of nano-formulating functional phytochemicals, showing promising scolicidal activity in vitro. The developed formulations, especially Nano-GL, showed the characteristics of an ideal scolicidal agent. Further studies are needed to assess in vivo efficacy and safety of this formulation

Scolicidal and apoptotic effects of phyto- and chemically synthesized silver/boehmite nanocomposites on Echinococcus granulosus protoscoleces

Cystic hydatid disease (CHD) is a zoonotic disease caused by the larval stage of Echinococcus granulosus (E. granulosus). This study aimed to synthesize silver nanoparticles (Ag NPs), silver boehmite nanocomposite (Ag/Bhm NC), and silver boehmite nanocomposite modified with chitosan (Ag/Bhm/Chit NC) using Rosmarinus officinalis (R. officinalis) extract and chemical method, and to evaluate their scolicidal and apoptotic effects on protoscoleces (PSCs) in vitro. The nanomaterials (NMs) were characterized by XRD, FTIR, FESEM, EDS, DLS, PDI, and zeta potential (ZP). The NMs were tested against PSCs at different concentrations (0.2–1.6 mg/mL) and exposure times (10–60 min). The size of Ag NPs, phytosynthesized Ag/Bhm NC, Ag/Bhm/Chit NC, and chemically synthesized Ag/Bhm NC were 25.55, 43, 72.3, and 60.8 nm, respectively. Ag NPs and phytosynthesized Ag/Bhm NC showed the highest scolicidal effect, with 65.34 % and 51.60 % mortality rate at 1.6 mg/mL and 60 min, respectively. Caspase-3 mRNA expression was higher in PSCs treated with Ag NPs and Ag/Bhm NC than in control groups (P < 0.05). Phytosynthesized Ag/Bhm NC had stronger scolicidal and apoptotic effect than chemically synthesized Ag/Bhm NC. Ag/Bhm/Chit NC had a weaker scolicidal effect but higher gene expression than Ag/Bhm NC. In conclusion, this study demonstrates the potential of phytosynthesized Ag NPs and Ag/Bhm NC as effective scolicidal and apoptotic agents against PSCs of hydatid cysts, which may be useful for the treatment of this disease.

In vitro and in silico scolicidal effect of sanguinarine on the hydatid cyst protoscoleces.

We aimed to investigate the scolicidal effects of sanguinarine on hydatid cyst protoscoleces (PSCs) in vitro and in silico. Different targets were docked into the active sites of sanguinarine. Molecular docking processes and visualization of interactions were performed using AutoDock Vina and Discovery Studio Visualizer. Binding energy was calculated and compared (kcal/mol). PSCs were aspirated from the hydatid cysts and washed. The sediments of PSCs were then exposed to various concentrations (50, 25, 12, 6, 3, and 1 μg/mL) of sanguinarine. The viability test was finally evaluated by the Trypan blue solution 4%. Levels of malondialdehyde (MDA), superoxide dismutase (SOD), glutathione (GSH), glutathione peroxidase (GPX), and catalase were analyzed to assess the level of oxidative stress-treated PSCs. Caspase-3 activity rate was determined to evaluate cell apoptosis in treated PSCs. Among the receptors, acetylcholinesterase was identified as the excellent target, with Vina score of -11.8. Sanguinarine showed high scolicidal effects after 12, 24, and 48 h. Also, in the first hour of exposure to the drug, caspase-3 activity and MDA level significantly increased, but the levels of GSH and GPx had a significant reduction after 12, 24, and 48 h (P < 0.05). The findings of this study revealed that sanguinarine have potent scolicidal effects in vitro and in silico and could be considered an opportunity for the introduction of a novel and safe therapeutic agent for the treatment of cystic echinococcosis. However, supplementary studies will be desired to prove the current findings by examining sanguinarine in a clinical setting.

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.

Protoscolicidal effects of curcumin nanoemulsion against protoscoleces of Echinococcus granulosus

BackgroundThe aim of the present study was to assess in vitro protoscolicidal effects of curcumin nanoemulsion (CUR-NE) against protoscoleces of cystic echinococcosis (CE)/hydatid cysts.MethodsThe CUR-NE was prepared via spontaneous emulsification of soybean as the oil phase, a mixture of Tween 80 and Tween 85 as the surfactant, ethanol as the co-surfactant and distilled water. Various concentrations of CUR-NE (156, 312, 625 and 1250 µg/ml) were exposed to collected protoscoleces of infected sheep liver hydatid cysts for 10, 20, 30, 60 and 120 min. Viability of the protoscoleces were assessed using eosin exclusion test. Morphological changes of the protoscoleces were observed using differential interference contrast (DIC) microscopy.ResultsThe mean particle size and zeta potential of CUR-NE included 60.4 ± 14.8 nm and − 16.1 ± 1.1 mV, respectively. Results showed that the viability of the protoscoleces decreased significantly with increases in CUR-NE concentrations (p < 0.001). The mortality rates of protoscoleces with exposure to concentrations of 1250 and 625 µg/ml of CUR-NE for 60 min were 94 and 73.33%, respectively. Mortality of the protoscoleces was 100% after 120 min of exposure to 1250 and 625 µg/ml concentrations of CUR-NE. Using NIC microscopy, extensively altered tegumental surface protoscoleces was observed after protoscoleces exposure to CUR-NE.ConclusionThe findings of the present study revealed the in vitro protoscolicidal potential of CUR-NE. Therefore, CUR-NEs are addressed as novel protoscolicidal agents, which can be used as an alternative natural medicine to kill the protoscoleces, owing to their low toxicity and significant inhibition potency. However, further studies are necessary to investigate pharmacologic and pharmacokinetics of CUR-NEs.

Antiparasitic Effects and Cellular Mechanisms of Formononetin (a Natural Isoflavone) Against Hydatid Cyst Protoscoleces

Background: The chemical agents applied to reduce the complications of hydatid cyst surgery are not free of side effects. Formononetin (FMN), as a natural isoflavone, has been shown to have various therapeutic benefits. Objectives: We studied the in vitro and ex vivo protoscolicidal activity and cellular mechanisms of FMN against hydatid cyst protoscoleces. Methods: Formononetin at 75, 150, and 300 μg/mL was mixed with hydatid cyst protoscoleces (103/mL), and the viability was determined in 5 to 60 min through eosin staining assay. The effect of FMN on caspase-3 activity was tested through a commercial colorimetric protease kit. In addition, the penetrability of the plasma membrane of protoscoleces after exposure to FMN was also determined by the SYTOX assay. Results: Formononetin at 300 μg/mL completely destroyed hydatid cyst protoscoleces after 30 minutes. While at 150 μg/mL, after 60 minutes of contact, 100% of protoscoleces were eliminated. By ex vivo assay, FMN showed its antiparasitic effect for longer periods. Based on these results, FMN at 300 μg/mL concentration completely eliminated hydatid cyst protoscoleces after 60 minutes of exposure. While at a concentration of 150 μg/mL after 60 minutes, 89.6% of protoscoleces were destroyed. Formononetin noticeably increased (P &lt; 0.001) the activity of caspase-3 and the permeability of protoscoleces dose-dependently. Conclusions: Formononetin as a natural product showed promising effects on the protoscoleces of hydatid cysts, indicating that it can be considered a valuable scolicidal agent. However, additional investigations are necessary to evaluate its efficacy in animal models and human subjects.

Comparison of the Scolicidal Activity of Pulicaria gnaphalodes and Alhagi maurorum Extracts against Protoscoleces of Echinococcus granulosus Sensu Lato In Vitro

Background: Cystic echinococcosis (CE), caused by the larval stage of the Echinococcus granulosus, is a zoonotic disease and has a global distribution. Today, herbal compounds are highly regarded in order to inactivate hydatid cyst protoscoleces. This study aimed to compare the scolicidal activity of hydroalcoholic extract of Pulicaria gnaphalodes and Alhagi maurorum against hydatid cyst protoscoleces in vitro. Methods: The scolicidal activity of P. gnaphalodes and A. maurorum extracts were evaluated at 50, 100, 150, and 200 mg/mL concentrations following 15, 30, and 60 minutes of exposure. Then, they were compared with Albendazole (5 g/100 mL) as positive control and distilled water as negative one in similar doses. The viability of protoscoleces was confirmed with a 0.1% eosin stain test under a light microscope. The experiments were performed twice, and data were analyzed by GraphPad software version 5.0. Results: The results of this study indicated that P. gnaphalodes extract killed 100% of the protoscoleces at a concentration of 200 mg/mL after 30 minutes of exposure, but the hydroalcoholic extract of A. maurorum at the same concentration and time could kill 90% of protoscoleces. Conclusion: The findings of the present study confirmed that P. gnaphalodes had a strong scolicidal effect; however; in vivo studies are needed to evaluate the effectiveness of P. gnaphalodes plant.

High potency of magnetic iron oxide nanoparticles covered by piroctone olamine against cystic echinococcosis

This study examined the synthesis of magnetic iron oxide nanoparticles coated with PO (FOMNPsP) and assessed their in vitro, ex vivo, and in vivo effects against cystic echinococcosis. The FOMNPsP was synthesized through the alkalization of iron ions in a deoxygenated form. In vitro and ex vivo protoscolicidal effects of FOMNPsP (100–400 μg/mL) were evaluated on hydatid cyst protoscoleces by the eosin exclusion test for 10–60 min. The effect of FOMNPsP on caspase-3 gene expression and exterior ultra-structural of protoscoleces was assessed by real-time PCR and scanning electron microscopy (SEM), respectively. In vivo effects were assessed by evaluating the number, size, and weight of hydatid cysts among infected mice. The FOMNPsP size was < 55 nm, and the most frequent particles were in the 15–20 nm range. In vitro and ex vivo assays revealed that the highest protoscolicidal effect was observed at 400 µg/mL with 100% lethality. After exposure of protoscoleces with FOMNPsP, the level of gene expression of caspase-3 was dose-dependently increased (p < 0.05). By SEM, the FOMNPsP-treated protoscoleces showed wrinkles and bulges resulting from the formation of blebs. FOMNPsP significantly decreased (p < 0.01) the mean number, size, and weight of the hydatid cyst. FOMNPsP revealed the potent protoscolicidal traits through disrupting the cell wall and apoptosis induction. The results also indicated the promising effect of FOMNPsP in controlling hydatid cysts in the animal model. Although FOMNPsP is safe for human normal cells, more investigations are required to clarify its toxicity and precise mechanisms of action.

Cytotoxic, Scolicidal, and Insecticidal Activities of Lavandula stoechas Essential Oil

Essential oils (EOs) have recently attracted more interest due to their insecticidal activities, low harmfulness, and rapid degradation in the environment. Therefore, Lavandula steochas (L. steochas) essential oil was assessed for its chemical constituents, in vitro cytotoxicity, and scolicidal, acaricidal, and insecticidal activities. Using spectrometry and gas chromatography, the components of L. steochas EOs were detected. Additionally, different oil concentrations were tested for their anticancer activities when applied to human embryonic kidney cells (HEK-293 cells) and the human breast cancer cell line MCF-7. The oil’s scolicidal activity against protoscolices of hydatid cysts was evaluated at various concentrations and exposure times. The oil’s adulticidal, larvicidal, and repelling effects on R. annulatus ticks were also investigated at various concentrations, ranging from 0.625 to 10%. Likewise, the larvicidal and pupicidal activities of L. steochas against Musca domestica were estimated at different concentrations. The analyses of L. steochas oil identified camphor as the predominant compound (58.38%). L. steochas oil showed significant cytotoxicity against cancer cells. All of the tested oil concentrations demonstrated significant scolicidal activities against the protoscoleces of hydatid cysts. L. steochas EO (essential oil) showed 100% adulticidal activity against R. annulatus at a 10% concentration with an LC50 of 2.34%, whereas the larvicidal activity was 86.67% and the LC50 was 9.11%. On the other hand, the oil showed no repellent activity against this tick’s larva. Furthermore, L. steochas EO achieved 100% larvicidal and pupicidal effects against M. domestica at a 10% concentration with LC50 values of 1.79% and 1.51%, respectively. In conclusion, the current work suggests that L. steochas EO could serve as a potential source of scolicidal, acaricidal, insecticidal, and anticancer agents.

Antiparasitic activity of Astragalus brachycalyx subsp. brachycalyx extract against hydatid cyst protoscoleces and its effect on induction of apoptosis: an in vitro and ex vivo study

Introduction: This study aims to evaluate in vitro and ex vivo antiparasitic activity of Astragalus brachycalyx subsp. brachycalyx. root chloroformic extract against hydatid cyst protoscoleces and its effect on induction of apoptosis. Methods: Various concentrations of the A. brachycalyx root chloroformic extract (56.25, 112.5, 225, and 450 mg/mL) were treated with hydatid cyst protoscoleces collected from the liver of infected sheep for 5-60minutes in vitro and ex vivo. Eosin exclusion test was also utilized to measure the mortality of protoscoleces. Moreover, the extract effect was assessed on apoptosis induction in hydatid cyst protoscoleces by caspase-3 activity measurement. Results: The mortality rate of protoscoleces in in vitro was 100% after being exposed to 450 and 225 mg/mL of A. brachycalyx extract for 20 and 30 minutes and in ex vivo for 30 and 60 minutes, respectively. Following 48 h treatment of protoscoleces, A. brachycalyx chloroformic extract at the doses of 56.25, 112.5, 225, and 450 mg/mL, dose-dependently motivated the caspase-3 enzyme ranging from 8.8% to 29.6% Conclusion: A. brachycalyx root chloroformic extract had a significant protoscolicidal effect; however, extra surveys are required to assess its efficacy and safety as a promising protoscolicidal agent in clinical settings.

Effects of green synthesized zinc nanoparticles alone and along with albendazole against hydatid cyst protoscoleces

BackgroundThe current investigation aims to green synthesized the zinc nanoparticles (ZnNPs) using Lavandula angustifolia extract by microwave technique and its protoscolicidal effects alone and combined with albendazole against hydatid cyst protoscoleces. MethodsDifferent concentrations of the ZnNPs (50, 100, and 200 μg/ml) alone and combined with albendazole (ALZ, 100 μg/ml) were treated with hydatid cyst protoscoleces obtained from liver of infected sheep for 5–60 min in vitro and ex vivo. Eosin exclusion examination was used to assess the viability of protoscoleces. The induction of apoptosis in hydatid cyst protoscoleces was assessed by measurement of the Caspase-3 activity of protoscoleces treated with various concentrations of ZnNPs. ResultsThe size of green synthesized ZnNPs was ranged from 30 to 80 nm, most of these nanoparticles were between 50 and 60 nm in size. In vitro, the highest scolicidal effect of ZnNPs was observed at the concentration of 200 μg/ml, where it killed 81.6% of protoscolices. While the combination of these nanoparticles with ALZ, especially at the concentration of 200 μg/ml, completely killed the protoscolices after 10 min’ exposure. However, compared to in vitro assay, the drugs tested took longer to show their protoscolicidal effect. ConclusionBased on the obtained results, ZnNPs particularly in combination with albendazole displayed the potent protoscolicidal in vitro and ex vivo as an intraperitoneal model of administration of agents to hydatid cyst treatment; nevertheless, additional investigations are mandatory to evaluate the efficacy and safety Zn NPs as a favorable protoscolicidal agent in clinical setting.

In vitro toxicity of cinnamaldehyde and nanoemulsion of cinnamaldehyde on protoscoleces of hydatid cyst.

Cystic echinococcosis is a major parasitic and zoonotic disease and surgery is the most common treatment of this disease which carries the risk of intraoperative leakage and recurrence. Using scolicidal agent to inactivate cyst contents reduces the risk of recurrence. Considering side effects of available scolicidals and growing interests on natural pharmaceuticals, the present study aimed to evaluate toxicity of cinnamaldehyde (CA), the main component of cinnamon essential oil, and a developed nanoemulsion of cinnamaldehyde (nano-CA) on protoscoleces of hydatid cyst. Nanoemulsion was prepared by the low energy system and characterized by dynamic light scattering to confirm dimensions. For evaluation of scolicidal effects, serial dilutions of CA and nano-CA were mixed with protoscolices suspension and mortality were recorded at 10, 30, and 60 minutes by eosin exclusion test. Albendazole was used as the positive control. The mean diameter of nano-CA was characterized as equal to 88.5 nm, and poly dispersity index was 0.09. After 30 min of treatment, nano-CA, at 50 μg/ml, killed 99.33% of protoscoleces. At the same time point and concentration, CA only caused mortality rate of 26.18%. 30 min-LC50 value of 369.39 μg/ml was obtained for CA, while after 30 min of exposure, nano-CA showed promising rapid activity with LC50 value of 3.22 μg/ml. Nano formulation significantly increased scolicidal activity of CA probably by increasing penetration and tegumental disorganization of protoscoleces. Further in vivo safety studies are needed to introduce nano-CA as a clinically applicable scolicidal agent.

In vitro scolicidal activity of clove oil and its nano-emulsion on hydatid cyst protoscoleces.

In vitro scolicidal activity of clove oil and its nano-emulsion on hydatid cyst protoscoleces.