Anti-echinococcal effects of sumac, Rhus coriaria, in a murine model of cystic echinococcosis: Parasitological and molecular evaluation.

Sumac has been traditionally used by people as a medicinal plant for the treatment of different disorders. Cystic echinococcosis (CE) is one of the major zoonotic diseases of human with a worldwide distribution. Long term albendazole therapy is usually associated with side effects including impaired liver function and leukopenia. The present study investigated the efficacy of the methanolic extract of Sumac, Rhus coriaria, on the secondary hydatid cyst development in mice and evaluated sumac effects on the expression of a profile of genes with a potential role in parasite development. Thirty-six mice were intraperitoneally inoculated with of with 3000 protoscoleces and six months after induction of infection were divided into three groups that received either oral sumac extract, albendazole or distilled water. The mice were necropsied 45 days later and the volume and weight of cysts were measured. The expression level of five target genes were analyzed using RT-qPCR. The volume and weight of the cysts were significantly lower in the sumac group compared to the controls. Decreased expressions were found in four out of the five genes following sumac administration. While significantly lower expressions in the sumac group were found for the cdk6, b-raf, fgfr and ras genes, no significant difference was found in cdk2 expression as compared with the control groups. Findings of the present study indicate high efficacy of sumac on the size and volume of secondary hydatid cysts in a murine CE model. Further studies are required to explore the most active and effective ingredients of this natural product.

Ruta graveolens, Peganum harmala, and Citrullus colocynthis methanolic extracts have in vitro protoscolocidal effects and act against bacteria isolated from echinococcal hydatid cyst fluid.

Echinococcosis is a common and endemic disease that affects both humans and animals. In this study, the in vitro activities of methanolic extracts of Ruta graveolens, Peganum harmala aerial parts, and Citrullus colocynthis seeds against protoscolosis and isolated bacterial strains from hydatid cysts were assessed using disc diffusion methods and Minimum Inhibitory Concentration (MIC). The chemical composition of three methanolic extracts was studied using LC-MS. After 3 h of exposure to 40 mg/mL R. graveolens extract, a tenfold protoscolocidal effect was seen when compared to the convintional medication (ABZ) for the same duration (P < 0.05). The bacteria listed below were isolated from hydatid cyst fluid collected from a variety of sick locations, including the lung and liver. Micrococcus spp., E. coli, Klebsiella oxytoca, Enterobacter aerogenes, Enterobacter amnigenus, Pseudomonas aeruginosa, Staphylococcus xylosus, and Achromobacter xylosoxidans are among the bacteria that have been identified. The most effective extract was R. graveolens, followed by P. harmala and C. colocynthis, according to the results of antibacterial activity using the disc diffusion method. R. graveolens extract had the lowest MIC values (less than 2 mg/mL) against all microorganisms tested. This shows that the R. graveolens extract has additional properties, such as the ability to be both scolocidal and bactericidal. Because these bacteria are among the most prevalent pathogenic bacteria that increase the risk of secondary infection during hydatid cysts, the results of inhibitory zones and MICs of the R. graveolens methanol extract are considered highly promising.

Effect of Taxus baccata L. Extract on Hydatid Cyst Protoscolices In vitro.

Hydatidosis is the most important global parasitic infectious disease both in humans and animals, which can lodge at different organs of the host, such as liver, lung (even heart), and brain which may lead to death. Surgery is the main method for the treatment of hydatidosis. In surgical therapy of hydatidosis, the use of sporicidal agents is very important since these agents inactivate live protoscolices and prevent recurrence of infection. Presently, numerous scolicidal chemical agents have been administrated to inactivate the hydatid cyst contents. Recently, there has been a high tendency among researchers to evaluate and present herbal plants as alternative option due to inexpensiveness, availability, low side effects, and toxicity. This study aimed to evaluate the scolicidal effect of hydro alcoholic Taxus baccata L. extract in vitro for the first time. The scolicidal activities of the extract were tested in concentrations of 50, 100, and 150 mg/ml following 10, 30, and 60 min of incubation, and the experiments were performed in triplicate. Viability of protoscolices was confirmed by 0.1% eosin vital staining. The data were analyzed in SAS software (version 9.4). The results showed that the hydroalcoholic extract of Taxus baccata L. at the concentration of 150 mg/ml led to killing 66.6% of protoscolices at 60 min. according to the results of this investigation, it is recommended to use this plant as a scolicidal plant. The findings of the present study showed that Taxus baccata L. had potent scolicidal effects. However, further studies are required to evaluate the efficacy of Taxus baccata L. in vivo.

Copper nanoparticles: Biosynthesis, characterization, and protoscolicidal effects alone and combined with albendazole against hydatid cyst protoscoleces.

BACKGROUND: Surgery remains the preferred treatment option for hydatid cyst (cystic echinococcosis); however, recent studies have demonstrated that the current protoscolicidal agents used during surgery are associated with some adverse side effects such as biliary fibrosis, hepatic necrosis, and cirrhosis. The present study aims to evaluate the in vitro and ex vivo anti-parasitic effects of copper nanoparticles (CuNPs) alone and combined with albendazole on hydatid cyst protoscoleces. METHODS: CuNPs was green synthesized using C. spinosa extract. Various concentrations of CuNPs (250, 500, and 750 mg/mL) alone and combined with albendazole (ALZ, 200 mg/mL) were exposed to protoscoleces collected from the liver fertile hydatid cysts of infected sheep for 5-60 min in vitro and ex vivo. Next, the eosin exclusion test was applied to determine the viability of protoscoleces. Caspase-3 like activity of CuNPs-treated protoscoleces was then evaluated using the colorimetric protease assay Sigma Kit based on the manufacturer's instructions. RESULTS: Scanning electron microscopy (SEM) results showed that the particle size of CuNPs was 17 and 41 nm with the maximum peak at the wavelength of 414 nm. The maximum protoscolicidal activity of CuNPs was observed at the concentration of 750 mg/mL in vitro, so that 73.3 % of protoscoleces were killed after 60 min of exposure. Meanwhile, the mortality of protoscoleces was 100 % after 10 min of exposure to 750 mg/mL of CuNPs along with ALZ (200 mg/mL). Nevertheless, the findings proved that CuNPs even in combination with ALZ required a longer time to kill protoscoleces ex vivo. After 48 h of treating protoscoleces, CuNPs in a dose-dependent manner and at doses of 250, 500, and 750 mg/mL induced the caspase enzyme activation by 20.5 %, 32.3 %, and 36.1 %, respectively. CONCLUSION: The findings of the present investigation showed potent protoscolicidal effects of CuNPs, especially combined with albendazole, which entirely eliminated the parasite after 10-20 min of exposure. The results also showed that although the possible protoscolicidal mechanisms of CuNPs are not clearly understood, the inducing apoptosis through caspases is one of the main protoscolicidal mechanisms of CuNPs. However, supplementary studies, especially in animal models and clinical settings, are needed to approve these results.

In vitro potential effect of Pipper longum methanolic extract against protoscolices of hydatid cysts.

This study aimed to evaluate the efficacy of methanolic extract of P. longum (PLM) against protoscolices of hydatid cyst in vitro. Four different concentrations of PLM extract (25, 50, 100 and 150 mg/ml) were used for the experiments. The metabolites in the PLM extract were characterized by Gas chromatography-mass spectrometry (GC-MS). The results showed the highest lethality of PLM extract in 50 mg/ml for 60 min exposure. The IC50 value obtained about 20 mg/ml for 60 min of PLM extract exposure. In this study, valuable findings were obtained for the first time about the scolicidal activity of P. longum, which is expected to conduct further studies in this field in the future.

High Potency of Organic and Inorganic Nanoparticles to Treat Cystic Echinococcosis: An Evidence-Based Review.

Since there is no potential, effective vaccine available, treatment is the only controlling option against hydatid cyst or cystic echinococcosis (CE). This study was designed to systematically review the in vitro, in vivo, and ex vivo effects of nanoparticles against hydatid cyst. The study was carried out based on the 06- PRISMA guideline and registered in the CAMARADES-NC3Rs Preclinical Systematic Review and Meta-analysis Facility (SyRF) database. The search was performed in five English databases, including Scopus, PubMed, Web of Science, EMBASE, and Google Scholar without time limitation for publications around the world about the protoscolicdal effects of all the organic and inorganic nanoparticles without date limitation in order to identify all the published articles (in vitro, in vivo, and ex vivo). The searched words and terms were: "nanoparticles", "hydatid cyst", "protoscoleces", "cystic echinococcosis", "metal nanoparticles", "organic nanoparticles", "inorganic nanoparticles, "in vitro", ex vivo", "in vivo". Out of 925 papers, 29 papers including 15 in vitro (51.7%), 6 in vivo (20.7%), ex vivo 2 (6.9%), and 6 in vitro/in vivo (20.7%) up to 2020 met the inclusion criteria for discussion in this systematic review. The results demonstrated the most widely used nanoparticles in the studies were metal nanoparticles such as selenium, silver, gold, zinc, copper, iron nanoparticles (n = 8, 28.6%), and metal oxide nanoparticles such as zinc oxide, titanium dioxide, cerium oxide, zirconium dioxide, and silicon dioxide (n = 8, 28.6%), followed by polymeric nanoparticles such as chitosan and chitosan-based nanoparticles (n = 7, 25.0%). The results of this review showed the high efficacy of a wide range of organic and inorganic NPs against CE, indicating that nanoparticles could be considered as an alternative and complementary resource for CE treatment. The results demonstrated that the most widely used nanoparticles for hydatid cyst treatment were metal nanoparticles and metal oxide nanoparticles, followed by polymeric nanoparticles. We found that the most compatible drugs with nanoparticles were albendazole, followed by praziquantel and flubendazole, indicating a deeper understanding about the synergistic effects of nanoparticles and the present anti-parasitic drugs for treating hydatid cysts. The important point about using these nanoparticles is their toxicity; therefore, cytotoxicity as well as acute and chronic toxicities of these nanoparticles should be considered in particular. As a limitation, in the present study, although most of the studies have been performed in vitro, more studies are needed to confirm the effect of these nanoparticles as well as their exact mechanisms in the hydatid cyst treatment, especially in animal models and clinical settings.

Subcutaneous Hydatid Cyst in Laboratory Mice: Is it a Suitable Method for Evaluating Therapeutic Agents against Hydatid Cyst?

Hydatid disease is an economic and public health concern in many countries. Currently, surgery is the main treatment option for hydatid disease. In the surgical treatment of hydatidosis, the use of scolicidal agents is very important due to inactivating live protoscoleces and preventing the recurrence of infection. Therefore, it is necessary to investigate newscolicidal agents and novel medications with higher safety and efficacy. In the previous in vitro studies, the scolicidal effects of the methanolic extracts and aromatic water of Zataria multiflora (Z. multiflora) have been demonstrated. Consequently, in this study, the impact of the nanoemulsion of Z. multiflora essential oil on subcutaneous hydatid cysts was compared with albendazole (ABZ). Fifty laboratory male mice were inoculated with 300 viable protoscoleces subcutaneously on the two sides of the abdomen. Following five months of infection, the remaining infected mice (n=42) were allocated into two treatment and one control (without treatment) groups containing fourteen animals each. Group A received ABZ at the dose of 50 mg/kg for 60 days, group B received the nanoemulsions of Z. multiflora at the dose of 50 mg/kg in drinking water for 60 days, and group C was considered as the control group. All the infected mice were euthanized and necropsied two months post-intervention. Afterwards, the cysts were cautiously collected and their number, size, and weight were compared between the mice of different groups. The mean number of hydatid cysts indicated that the nanoemulsion of Z. multiflora essence had a relative superiority to ABZ. On the other hand, the therapeutic effect of ABZ was higher than the nanoemulsion of Z. multiflora essential oil in terms of the mean weight and mean size of hydatid cysts. However, no significant difference was observed between the groups (P&gt;0.5). Overall, the number, weight, and size of cysts were not significantly different between the groups in this investigation. The lack of satisfactory therapeutic results in this study might be due to the location of hydatid cysts in the subcutaneous space.

Zataria Multiflora bois as an auspicious therapeutic approach against Echinococcus granulosus: Current status and future perspectives.

Hydatidosis caused by Echinococcus granulosus is a major zoonotic diseases. In addition to imposing heavy economic losses, the disease is a public health problem worldwide. The larval stage of the parasite (hydatid cyst) is formed in a wide range of domestic, wild and human beings as intermediate hosts. On the other, its recurrence has been reported anywhere as a reemerging disease. Although the cysts have some evading mechanisms, both human TH1 and TH2 cells subsets are stimulated. Because of increasing resistance and adverse effects of medications such as abnormalities of liver and other organs functions and abdominal pain, seeking alternative therapeutic approaches to be inexpensive, easy available, with low side effects and toxicity seems essential. However, the lack of information on the social and economic welfares of herbal medicines for the industrial scale application is a limitation. Zataria Multiflora bois (ZMB) has exhibited huge advantages and tremendous protoscolicidal effects as demonstrated by numerous studies and its combination therapies with anti-parasitic drugs have exerted desirable outcomes in vitro and in vivo. Noticeably, the compound confers negligible side effects or toxicity even at high concentrations. ZMB has exhibited promising inhibitory effects against hydatid cyst, particularly when combined with chemical drugs and in formulations of nanoemulsions. Its immunomodulatory effects include increase of nitric oxide production (NO) and protection of hepatic cells (Kupffer cells, fat-storing cells, and endothelial cells), enhancement of macrophages and T cells and increase of cytokines production. This study aimed at assessment of ZMB traits for application against hydatid cyst protoscolices.

Scolicidal activity of Mesobuthus eupeus venom against the protoscolices of Echinococcus granulosus.

Hydatidosis is an important zoonosis caused by a parasitic tapeworm, namely Echinococcus granulosus. This infection is distributed worldwide and affects the health as well as economic loss in both humans and animals. In most cases, the disease needs chemotherapy with or without surgery. Conventional drugs have some major problems, including drug complications, harmful side effects, and also progressive resistance. According to the importance of biological productions as alternative medicine, a large number of studies confirmed that whole venom and many peptide ingredients of the scorpion venom have various different medical benefits, including antimicrobial properties, due to the mechanism of blocking gated ion channel. In this study, the venom peptides of Mesobuthus eupeus scorpionwere purified using gel filtration chromatography and subsequently ion exchange chromatography, followed by the determination of the molecular weights of the proteins by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) procedure. After collecting the hydatid cysts fluids from the liver of infected sheep, protoscolices were derived, washed, and encountered to the whole venom as well as eight different fractions of toxin 30, 60, 120, and 240 min after the exposure. In the next step, the viability of protoscolices was determined by eosin staining. The obtained results revealed that a venom fraction under 10 kDa killed all protoscolices after 30 min. Moreover, it was found that the scolicidal activity of fractions increases according to the time of exposure. As a result, it can be concluded that M. epeus venom peptides under its LD50 (1/2 LD50) can properly and quickly destroy the protoscolices of hydatid cysts at the level of applied concentrations and such components are good alternatives to treat hydatidosis.

In-vitro evaluation of Nigella sativa and Punica granatum effect on protoscolices of hydatid cysts / Avaliação in vitro do efeito de Nigella sativa e Punica granatum em protoescólices de cistos hidáticos

Abstract Cystic echinococcosis (CE) are commonly found in the liver and lungs of affected hosts. The treatment approach is usually surgical, or giving drugs in conjunction before surgery to kill protoscolices, to avoid anaphylactic shock from leakage of hydatid fluid into the peritoneum and to decrease opportunities for recurrences. The present study was to evaluate the in vitro scolicidal efficacy of hydroalcoholic extract of Punica granatum peel and Nigella sativa, on the protoscolices of CE that collected from the lungs of infected camels. Different concentrations of extracts with different exposure times were used and a viability assay was applied to measure the scolicidal effect. N. sativa showed its highest scolicidal efficacy at 100 mg/mL and 10 mg/mL concentrations after 30 and 60 min. P. granatum peel extract showed its maximum scolicidal efficacy at 100 mg/mL concentration after 120 min. All experiments of the current study revealed that the extracts of both N. sativa and P. granatum had a scolicidal effects on the protoscolices of camel hydatid cysts. It could be concluded that N. sativa extract is more potent than P. granatum peel extract regarding scolicidal effect, but the efficacies of both extracts were of moderate significant correlation to exposure time and concentrations.

Resumo Os cistos hidáticos (equinococose cística, CE) são comumente encontrados no fígado e nos pulmões dos hospedeiros afetados. A abordagem do tratamento geralmente é cirúrgica, e algumas drogas são administradas em conjunto antes da cirurgia para matar protoscólices e evitar choque anafilático devido ao vazamento de fluido hidático no peritônio e diminuir as oportunidades de recorrência. O presente estudo foi avaliar a eficácia in-vitro do extrato hidroalcoólico de casca de Punica granatum e Nigella sativa, sobre os protoescólices de cistos hidáticos, que foram coletados dos pulmões de camelos infectados. Concentrações dos extratos com diferentes tempos de exposição foram utilizados e um ensaio de viabilidade foi aplicado para medir o efeito escolicida. A N. sativa apresentou sua maior eficácia escolicida nas concentrações de 100 mg/mL e 10 mg/mL após 30 e 60 min. O extrato de casca de P. granatum mostrou sua máxima eficácia escolicida na concentração de 100 mg/mL após 120 min. Todos os experimentos do presente estudo revelaram que os extratos de N. sativa e P. granatum tiveram efeito escolicida dependente da dose e do tempo nos protoescólices dos cistos hidáticos de camelo. Pode-se concluir que o extrato de N. sativa é mais potente que o extrato de casca de P. granatum em relação ao efeito escolicida, mas a eficácia de ambos os extratos foi de correlação significativa moderada com o tempo de exposição e as concentrações.

The scolicidal activity of the essential oil obtained from the needles of Pinus nigra Arn. subsp. pallasiana (Lamb.) Holmboe on hydatid cyst.

ETHNOPHARMACOLOGICAL RELEVANCE: This study aimed to evaluate the in vitro activity of the essential oil of Pinus nigra subsp. pallasiana against protoscoleces of Echinococcus granulosus sensu lato. In traditional Turkish folk medicine, the essential oils and resin of P. nigra and P. sylvestris are used externally for wound healing. They are also used in cattle for treating worms and as boiling essence against cold and stomachache. Protoscoleces were aseptically aspirated from sheep livers having hydatid cysts. MATERIALS AND METHODS: Three concentrations (10, 30, and 50 mg/mL) of the essential oil from P. nigra subsp. pallasiana cones were used for 10-60 min. The essential oil was applied to protoscoleces of hydatid cyst to determine the scolicidal activity of the essential oil. Viability of protoscoleces was observed by 0.1% eosin staining. RESULTS: The rate of dead protoscoleces was found to be 2.27% in the control group, which was not treated with essential oil. The rate of dead protoscoleces increased to 61.69%, 69.39%, 75.98% and 71.76% respectively after, 10, 20, 30, and 60 min when the protoscoleces were exposed to the P. nigra subsp. pallasiana essential oil at 10 mg/mL concentration. When protoscoleces were exposed to the essential oil of P. nigra subsp. pallasiana for 10, 20, 30, and 60 min at 30 mg/mL concentration, the mortality rates of protoscoleces were determined as 3.82%, 91.33%, 94.67% and 96.74%, respectively. The mortality rate of 100% was determined at 50 mg/mL concentration after 60 min of exposure. According to statistical analysis, P. nigra subsp. pallasiana essential oil at all concentrations had significant scolicidal effects compared to the control group at all exposure times. CONCLUSIONS: The essential oil of P. nigra subsp. pallasiana needles showed scolicidal activity and it could be a new scolicidal agent for using in hydatid cyst treatment.

In Vitro Study on Protoscolicidal Effect of Methanolic Extract of Allium hirtifolium on Protoscoleces of Cystic Echinococcosis.

BACKGROUND: Cystic echinococcosis/ hydatidosis is one of the most important parasitic zoonoses worldwide. Usually, surgery is used for the treatment of active cysts. Due to the risk of cyst rupture during surgery, a protoscolicidal agent is usually injected into the cyst before surgery to prevent secondary cyst formation in case of cyst rupture. Due to the side effects of chemical protoscolicidal agents attempts have been focused on herbal medicine. In this regard, Allium sativum has already been used; however, the protoscolicidal effect of Allium hirtifolium from Allium family has not been investigated, so far. OBJECTIVE: The present study was conducted in order to assess the protoscolicidal effect of methanolic extract of Allium hirtifolium in different concentrations, temperature and exposure times. Cetrimide as a positive control and DMSO as a negative control were also used in this study. METHODS: The viability of protoscoleces was determined by 0.1% eosin staining method. RESULTS: The protoscolicidal effect of Allium hirtifolium on protoscoleces was as follows: The mean percent and standard deviation (SD) of protoscolicidal activity of 50 mg/ml concentration of Allium hirtifolium extract in room temperature at 10, 20, 30, 40 and 50 minutes of exposure was 5.88±2.39, 8.59±4.54, 11.82±5.81, 12.31±6.16and 16.96±8.42 respectively; while at 37°C it was 9.10±5.01, 11.52±7.36, 18.64±4.36, 27.07±6.57 and 38.15±12.80, respectively. CONCLUSION: In conclusion, although Allium hirtifolium from Allium family is similar to Allium sativum; the protoscolicidal effect of its methanolic extract is not similar and less than A. sativum extract.

Allium Sativum Methanolic Extract (garlic) Improves Therapeutic Efficacy of Albendazole Against Hydatid Cyst: In Vivo Study.

After gas chromatography and mass spectrometry of prepared methanolic extract of Allium sativum, 40 laboratory BALB/c mice were infected intraperitoneally by injection of 1,500 viable protoscoleces. Five months after infection, the infected mice were allocated into four treatment groups, including 1- Albendazole (100 mg/kg); 2- Allium sativum methanolic extract (10 mL/L); 3- A. sativum methanolic extract (10 mL/L) + Albendazole (50 mg /kg); and 4- untreated control group. After 30 days of daily treatment, total number and weight of cysts and size of the largest cyst as well as blood serum bilirubin and liver enzymes were compared between the mice of different groups.The total number and weight of cysts and size of the largest cyst were significantly lower in treated groups A. sativum 10 mL/L + Albendazole 50 and Albendazole 100 in comparison to those of the control group (p < 0.05).The activity of alanine aminotransferase (ALT) enzyme and bilirubin concentration were significantly lower in the mice treated with A. sativum 10 mL/L and A. sativum 10 mL/L + Albendazole 50, when compared to the control group. In addition, bilirubin concentration revealed significant decrease in A. sativum 10 mL/L and A. sativum 10 mL/L + Albendazole 50 groups, when compared to the Albendazole group. In conclusion, administration of A. sativum 10 mL/L improved the anti-hydatidosis activity of Albendazole 50 mg /kg, due to parasitological effects similar to Albendazole 100 mg /kg but less hepatotoxic effects.

In vivo therapeutic efficacy of the Allium sativum ME in experimentally Echinococcus granulosus infected mice.

Plant-based anthelmintics suggest an alternative treatment for cystic echinococcosis. The present study was conducted to evaluate the efficacy of methanolic extract (ME) of A. sativum (garlic) on the treatment of hydatid cysts in the murine model. After gas chromatography and mass spectrometry of prepared ME, sixty laboratory BALB/c mice were infected intraperitoneally by injection of 1500 viable protoscoleces. Five months after infection, the infected mice were allocated into six treatment groups, 1- Albendazole (100 mg/kg); 2- Allium sativum ME (10 mL/L); 3- A. sativum ME (20 mL/L); 4- A. sativum ME (40 mL/L); 5- A. sativum ME (80 mL/L) and 6- untreated control group. After 30 days of daily treatment, the total number of cysts, size and weight of the largest cyst were significantly lower in three treated groups including A. sativum ME 80 mL/L, A. sativum ME 40 mL /L and albendazole in comparison to those of the control group (p < 0.05). The activity of alanine amino transferase (ALT) enzyme, as well as bilirubin concentration were significantly lower in the mice treated with A. sativum ME 80, 40, 20 and 10 mL/L when compared to the control group. In addition, bilirubin concentration revealed significant decrease in A. sativum ME 10, 20 and 80 mL/L groups, when compared to the albendazole group. In conclusions, administration of A. sativum ME used at 40 and 80 mL/L concentrations might be beneficial in the treatment of CE due to anti-parasitic effects similar to albendazole but less hepatotoxic effects.

Natural products applied against hydatid cyst protoscolices: A review of past to present.

Echinococcus granulosus is the causative agent of cystic echinococcosis (CE), which is distributed all around the world. CE is one of the most important global parasitic infectious diseases, both in humans and animals. This parasite causes hydatid cysts that can be lodge at different organs of host such as liver, lung even in heart and brain which may lead to death. Presently, numerous scolicidal chemical agents have been administrated for inactivation of the hydatid cyst contents. Because of increasing resistance and adverse effects of medications include abnormalities of liver function, abdominal pain, diarrhea, nausea, vomiting, dizziness, and headache; there is a need to find alternative therapies either with the least or without side effects. Recently, there is a high tendency among researchers to evaluate and present herbal plants as alternative option due to being inexpensive, easy available, low side effects and toxicity. Till now, many efforts have been conducted on herbal extracts against protoscolices of hydatid cysts throughout the world. Therefore, the current review systematically searched the following electronic databases: PubMed, Science Direct, Scopus, and Google Scholar on published papers according to the keywords. In addition, a comprehensive list of medicinal plants was prepared and some of these herbal plants which showed the best efficacy and promising results are discussed elaborately.

In vitro protoscolicidal effects of various concentrations of Ziziphora tenuior L. extract at different exposure times.

OBJECTIVE: Hydatidosis is one of the most important zoonotic diseases and surgery is still the main treatment for this problem. One of the side effects of hydatid cyst surgery is recurrence, thus, searching and assessment of some new agents such as medicinal plant extracts are very important. In the present study, the scolicidal effect of ethanolic extract of Ziziphora tenuior (Z. tenuior) was investigated. MATERIALS AND METHODS: Protoscolices were aseptically collected from sheep livers containing hydatid cyst and used in the experiments. Z. tenuior extract was used at concentration of 3-100 mg/ml for 10-60 min. Viability of protoscolices was determined by 0.1% eosin staining. RESULTS: Based on our results, Z. tenuior extract at concentration of 10 mg/ml killed all protoscolices after 20 min. However, this medicinal plant at concentration of 25 mg/ml destroyed all protoscolices in a shorter exposure time (10 min). Therefore, the scolicidal activity of the extract at 10 and 25 mg/ml concentrations was considerably effective in lower concentrations and shorter exposure times. CONCLUSION: The findings of this study showed that the ethanolic extract of Z. tenuior produces high scolicidal activity; it may be used as an appropriate and effective scolicidal agent in hydatidosis surgery. This is the first report on the protoscolicidal activity of Z. tenuior.

Chemical composition, efficacy and safety of Pistacia vera (var. Fandoghi) to inactivate protoscoleces during hydatid cyst surgery.

At present, various scolicidal agents have been used for inactivation of protoscoleces during hydatid cyst surgery, however, they are associated with serious adverse side effects including sclerosing colangititis (biliary tract fibrosis), liver necrosis and methaemoglobinaemia. This investigation was designed to evaluate the chemical composition and in vitro scolicidal effects of Pistacia vera (var. Fandoghi) essential oil against protoscoleces of hydatid cysts and also its toxicity in mice model. The components of the P. vera essential oil were identified by gas chromatography/mass spectroscopy (GC/MS) analysis. Protoscoleces were aseptically aspirated from sheep livers having hydatid cysts. Various concentrations of the essential oil (25-200µl/mL) were used for 5-30min. Viability of protoscoleces was confirmed using eosin exclusion test (0.1% eosin staining). In addition, forty male NIH mice were used to determine the acute and sub-acute toxicity of P. vera essential oil for 2 and 14 days, respectively. The main components of P. vera essential oil were limonene (26.21%), α-pinene (18.07%), α-thujene (9.31%) and α-terpinolene (9.28%). Findings of the present study demonstrated that the P. vera essential oil at the concentrations of 100 and 200µl/mL killed 100% protoscoleces after 10 and 5min of exposure, respectively. The LD50 values of intraperitoneal injection of the P. vera essential oil was 2.69ml/kg body weight, and the maximum nonfatal doses were 1.94ml/kg body weight. No significant difference (P>0.05) was observed in the clinical chemistry and hematological parameters following oral administrations of P. vera essential oil at the doses 0.1, 0.2, and 0.4ml/kg for 14 days. The obtained findings demonstrated new chemical composition and promising scolicidal activity of the P. vera with no significant toxicity which might be used as a natural scolicidal agent in hydatid cyst surgery.

Efficacy of Myrtus communis L. to Inactivate the Hydatid Cyst Protoscoleces.

PURPOSE: The present study aims to investigate the scolicidal effects of Myrtus communis L. essential oil against protoscoleces of hydatid cysts and also its toxicity in mice model. MATERIALS AND METHODS: Protoscoleces were aseptically aspirated from sheep livers having hydatid cysts. Various concentrations of the essential oil (12.5-100 µl/ml) were used for 5-30 min. Viability of protoscoleces was confirmed using eosin exclusion test (0.1% eosin staining). Moreover, 48 male NMRI mice were used to determine the acute and sub-acute toxicity of M. communis essential oil. One-way ANOVA with Tukey's post-hoc test was used to assess differences between experimental groups. RESULTS: Findings of the present study demonstrated that the M. communis essential oil at the concentration of 100 µl/ml after 5 min of exposure killed 100% protoscoleces. Similarly, the mean mortality rate of protoscoleces after 10 min of exposure to concentration of 50 µl/ml was 100%. However, lower concentrations (12.5 and 25 µl/ml) of M. communis essential oil provoked a delayed protoscolicidal effects. The LD50 values of intraperitoneal injection of the M. communis essential oil was 2.23 mL/kg body wt. No significant difference (p > .05) was observed in the clinical chemistry and hematological parameters following oral administrations of M. communis essential oil at the doses 0.05, 0.1, 0.2, and 0.4 mL/kg for 14 days. CONCLUSION: The results showed potent scolicidal activity of M. communis with no significant toxicity, which might be used as a natural scolicidal agent in hydatid cyst surgery.

Chemical composition, protoscolicidal effects and acute toxicity of Pistacia atlantica Desf. fruit extract.

The present study was designed to evaluate the chemical composition and scolicidal effects of Pistacia atlantica Desf. extract against protoscoleces of hydatid cysts and its acute toxicity in mice model. Various concentrations of the methanolic extract (5-50 mg/mL) were used for 10-60 min. Viability of protoscoleces was confirmed using eosin exclusion test (0.1%). Acute toxicity was also determined in mice model. The main components were ß-myrcene (41.4%), α-pinene (32.48%) and limonene (4.66%). Findings demonstrated that P. atlantica extract at the concentrations of 25 and 50 mg/mL after 20 and 10 min of exposure killed 100% protoscoleces. The LD50 of the intraperitoneal injection of the P. atlantica methanolic extract was 2.43 g/kg and the maximum non-fatal dose was 1.66 g/kg. Obtained results showed the potential of P. atlantica extract as a natural source with no significant toxicity for the production of new scolicidal agent to use in hydatid cyst surgery.

Preventive and therapeutic effects of Zataria multiflora methanolic extract on hydatid cyst: an in vivo study.

The phenolic compounds of Zataria multiflora extract, were identified by HPLC analysis. Gallic acid, catechin, caffeic acid, and quercetin were found to be the major phenolic compounds. Eighty healthy laboratory Balb/C mice were infected intraperitoneally by injection of 1500 viable protoscoleces and were divided into prevention (40 mice) and therapeutic (40 mice) groups. To prove the preventive effect of Z. multiflora extract on development of hydatid cyst, the 40 infected animals were allocated into three treatment groups including Z. multiflora (4 g/l in drinking water for 8 months), albendazole (150 mg/kg BW/day for 10 days) and untreated (control) group. To estimate the therapeutic effect of Z. multiflora extract on the hydatid cyst, after 8 months of infection, the infected mice were allocated into three experimental treatment groups including Z. multiflora (8 g/l in drinking water for 30 days), albendazole (300 mg/kg BW/day for 20 days) and untreated (control) group. At the end of the treatment period, all mice were euthanized and necropsied, the hydatid cysts were carefully removed, weighed and their size were recorded. Weight and size of the hydatid cysts significantly decreased (p<0.05) upon the treatment with Z. multiflora extract in both prevention and therapeutic groups. The germinal layer of the hydatid cysts recovered from the treated mice, either from the prevention or therapeutic group, were completely damaged at ultrastructural level by scanning electron microscopy.