Rapid Detection and Identification of Fasciola spp. and Dicrocoelium spp. Isolated from the Ruminant Livestock of Northwest Iran Using High-Resolution Melting Analysis (HRM).

Background: The liver flukes of the Fasciola species and Dicrocoelium spp. are recognised as parasites of domestic and wild herbivores. Both species of F. hepatica and F. gigantica as well as D. dendriticum are distributed in Iran. The present study aimed to identify Fasciola spp. and Dicrocoelium spp. using mitochondrial Cox1 (cytochrome c oxidase I) gene by HRM method. Methods: Totally, thirty infected liver specimens were collected from the sheep (n:23) and cattle (n:7) at the abattoirs of Qazvin Province, northwest Iran in 2022. DNA extraction and PCR amplification of Cox1 gene were conducted by HRM technique. DnaSP v.5.0 was used for compression of diversity indices of ribosomal 28S rDNA and mitochondrial Cox1 markers of Dicrocoelium spp. The taxonomic status of Dicrocoelium spp. was performed by sequencing and phylogenetic analysis. Results: Overall, 26 and 4 isolates were identified as F. hepatica and F. gigantica, respectively. D. dendriticum was the sole infecting species of Dicrocoelium revealed by HRM analysis. Genomic analysis showed a moderate (28S rDNA genes: 0.600±0.215) to high (Cox1: 0.733±0.155) haplotype diversity for D. dendriticum. Conclusion: The parasite-dependent mitochondrial gene (Cox1) could identify a higher genetic diversity of D. dendriticum compared to nuclear 28S rDNA gene. HRM technique in the present study found to be a reliable technique for identification and genetic diversity of liver flukes but more comprehensive and in-depth studies in different parts of the country are needed.

In vitro anti-Toxoplasma gondii activity of Ganoderma lucidum extracts.

OBJECTIVE: Ganoderma extracts have the potential to be used as anti-cancer, anti-inflammatory, immunomodulator, and antimicrobial agents, as evaluated in numerous studies. This study was aimed to determine the lethal and inhibitory effects of aqueous, hydroalcoholic, and alcoholic extracts of Ganoderma lucidum on Toxoplasma gondii RH strain tachyzoites, in vitro. RESULTS: All three types of extracts showed toxoplasmacidal effects. The highest percentage of mortality was related to hydroalcoholic extract. The EC50 of Ganoderma extracts for tachyzoites were 76.32, 3.274, and 40.18 for aqueous, hydroalcoholic and alcoholic extracts, respectively. The selectivity index obtained for hydroalcoholic extract was 71.22, showing the highest activity compared to other extracts. According to our findings, the hydroalcoholic part was the most effective substance among the extracts. This basic study showed obvious anti-toxoplasma effect of Ganoderma lucidum extracts. These extracts can be used as candidates for further in-depth and comprehensive studies especially In vivo experiments to prevent toxoplasmosis.

Isolation and Identification of Free Living Amoeba from Patients and Contact Lens Users in Iran.

Background: Free-living amoebae (FLA) such as Acanthamoeba spp., are considered as opportunistic and pathogenic protozoans. Acanthamoeba granulomatous encephalitis (AGE) is a serious threat for immunodeficient patients and Acanthamoeba keratitis (AK) for contact lens users. We aimed to identify the presence of free living amoebae in nasal swabs of patients and contact lens users in Qazvin, Iran. Methods: During 2019, 251 nasal and oral swabs (including the pharynx and mouth) were collected from patients with diabetes, AIDS and those under periodic dialysis in Qazvin, Iran. In addition, 27 soft contact lenses were collected from the participants. Following DNA extraction, PCR and sequencing were conducted to identify the genotypes of the amoeba. Phylogenetic analysis of the identified sequences was performed using MEGA 7 software. Results: A strain of Acanthamoeba belonging to the T3 genotype was isolated from hemodialysis patients. Two specimens of Acanthamoeba with T3 genotype were isolated from keratitis patients. Conclusion: The clinicians should pay attention to the possible complication of this organism because this amoeba is potentially pathogenic for immunocompromised patients. Since the amoeba is present in environmental resources, the use of contact lenses should be accompanied by considering proper hygiene.

Molecular identification and phylogenetic analysis of free-living amoeba in the water resources of Arak, Iran.

The aim of the present study was to detect free-living amoeba (FLA) in the water resources of Arak, Iran using molecular tools. A total of 154 samples were collected from different water supplies. Molecular analyses, sequencing, and phylogenetic study were conducted to confirm the species and genotypes of FLA. Fisher exact test was used to determine the significance. Of 154 water samples, 19 (12.3%) samples were tested positive for FLA. Three genotypes of Acanthamoeba including T4, subtype D, and T5 were identified among the isolates. The pathogenicity assay showed that the isolate of Acanthamoeba in drinking water was highly pathogenic. Three species of Naegleria, including N. australiensis, N. pagei, and N. gruberi were found among the samples. Six isolates of Vermamoeba were identified as V. vermiformis. Meanwhile, three other species including Vannella sp., Vahlkampfia avara, and Stenamoeba polymorpha were also recovered from the water samples. Statistical analysis showed a significant difference between the various water resources contaminated with FLA. This is the first study to reveal the presence of S. polymorpha in water sources in Iran. According to the findings of the present study, health officials should be beware of potential public health impacts of FLA in water resources.

Cutaneous leishmaniasis as an increasing threat for Iranian travellers attending religious ceremonies.

BACKGROUND: The Islamic Republic of Iran is one of the most important endemic foci of cutaneous leishmaniasis in the world. Annually, a large number of Iranian Shia pilgrims travel to Iraq from this area in order to participate in one of their most important religious ceremonies. This trip has coincided with the seasonal activity of sand flies in recent years. So, cutaneous leishmaniasis could be a serious threat for pilgrims on these trips. AIMS: To report cases of cutaneous leishmaniasis among Iranian Shia pilgrims attending a religious ceremony in Iraq during 2017. METHODS: Sixteen patients were referred to our laboratory in the Department of Parasitology and Mycology at Qazvin University of Medical Sciences. Dermal scrapings and stained slides prepared of skin lesions were used to morphological diagnosis. DNA extraction and PCR amplification were optimized to identification of Leishmania species. RESULTS: All of the patients were infected with cutaneous leishmaniasis in microscopic survey. L. major was detected by molecular approach. The number of lesions observed in patients were 1 (31%), 2 (25%), and ≥ 3 (44%). CONCLUSIONS: Since a large number of Shia Muslims participate in the annual religious ceremonies, serious measures must be taken to prevent the disease.

Molecular Characterization of Echinococcus granulosus Sensu Stricto Isolated from the Livestock of Qazvin, Iran.

INTRODUCTION: Hydatidiosis is a serious parasitic disease in humans and livestock, worldwide. Echinococcus granulosus shows notable genetic variation among intermediate hosts. Several genotypes of the worm have been reported from different parts of Iran, but no information on the parasite genotypes status in the study region is available. The current study investigated the presence of different genotypes of E. granulosus in the livestock of Qazvin, Iran, by sequencing the mitochondrial Cox1 genes. METHODOLOGY: One hundred twenty E. granulosus isolates, including 30 from goats, 40 from cattle and 50 from sheep, were collected from the slaughterhouses in Qazvin province. Mitochondrial Cox1 gene region was amplified by PCR and 30 isolates were sequenced. Phylogenetic analysis was performed by using the MEGA 7.0 software. Morphological analysis was performed on rostellar hook length of protoscoleces. RESULTS: All isolates were identified as E. granulosus sensu stricto (G1-G3 complex) among 17% of the isolates clarified as G3 genotypes. G1 was the predominant genotype among the specimens. No significant difference between the rostellar hooks measurements of different genotypes was observed. CONCLUSION: Our findings confirmed the presence of E. granulosus sensu stricto in the region, although further studies are required to determine the haplotype diversity of E. granulosus using different mitochondrial and nuclear genes.

Isolation and Molecular Identification of Acanthamoeba and Naegleria from Agricultural Water Canal in Qazvin, Iran.

BACKGROUND: Free-living amoeba (FLA) are widely distributed in different environmental sources. The most genera of the amoeba are Acanthamoeba, Naegleria and Vermamoeba. The most common consequences of the infections in immune-deficient and immuno-competent persons are amoebic encephalitis and keratitis. The aim of this study was to investigate the presence of Acanthamoeba spp. and Naegleria spp., isolated from the main agricultural water canal in Qazvin. METHODS: Totally, 120 water specimens were collected and later the specimens were cultured and cloned to identify positive samples. PCR amplification and sequencing were carried out to identify the isolated species as well as the genotypes of amoeba. RESULTS: According to morphological surveys, 41.7% (50/120) of water specimens were positive for FLA. Molecular analysis revealed that 68.6% and 31.4% of Acanthamoeba specimens were identified as T3 and T4 genotypes, respectively. Also, two species of Naegleria named as N. lovaniensis (57.1%) and Naegleria sp. (42.8%) were identified. The results of pathogenicity assays demonstrated that 38.5% of T3 and 61.5% of T4 genotypes of Acanthamoeba were highly pathogenic parasites. CONCLUSION: The water flowing in the agricultural canal of the area is contaminated with potential pathogenic FLA, therefore, it is recommended that more attention to be paid towards proper treatment of water sources to prevent possible risk of the disease.

Prevalence of Intestinal Parasites among Rural Inhabitants of Fouman, Guilan Province, Northern Iran with Emphasis on Strongyloides stercoralis.

BACKGROUND: Intestinal parasitic infections (IPIs) are among the most important etiologies of gastrointestinal disorders in developing countries. The present study was performed to determine the prevalence of IPIs in rural inhabitants of Fouman, northern Iran. METHODS: Overall, 31 villages were randomly selected during 2015-2016. Stool samples were collected from 1500 inhabitants aged 2-87. The samples were examined by direct wet smear, formalin ethyl-acetate concentration and agar plate culture. Trichrome staining and modified acid-fast staining were used as confirmatory tests for intestinal amoeba and flagellates and cryptosporidium spp., respectively. Data were analyzed with Chi-Square and Fisher exact tests using SPSS. RESULTS: 8.06% of participants were positive for at least one intestinal parasite. The prevalence of mixed parasitic infections was 0.87%. The most prevalent IPIs were caused by Trichostrongylus spp. (3.13%), followed by Strongyloides stercoralis (1.5%), Giardia lamblia (1.3%), and Entamoeba coli (1.0%), Blastocystis hominis (0.86%), E. histolytica/dispar (0.53%), Endolimax nana (0.26%), Iodamoeba butschlii (0.13%), Trichuris trichiura (0.07%), Enterobius vermicularis (0.07%), Hook worm (0.07%) and E. hartmani (0.07%). Statistically, the prevalence of IPIs showed significant differences regarding the age groups, education status, occupation (P<0.001), and the habit of eating raw vegetables (P<0.007), whereas, the differences were insignificant with regard to sex (P=0.924) and water supply (P=0.088). CONCLUSION: The prevalence of IPIs, especially soil-transmitted helminthes (STHs) has sharply decreased in northern Iran. Excluding Trichostrongylus spp. and S. stercoralis, other intestinal parasites only produce a marginal and unnoticeable health problem in this area, today.

Isolation and identification of Acanthamoeba genotypes and Naegleria spp. from the water samples of public swimming pools in Qazvin, Iran.

Free-living amoeba (FLA), including Acanthamoeba and Naegleria are facultative parasites in humans. The amoeba have widespread distribution in various water sources. The aim of this study was isolation and molecular identification of Acanthamoeba and Naegleria isolated from swimming pools and also hot and cold tub waters in Qazvin province. The samples (166 water samples) were cultured to isolate and identify positive specimens. PCR (polymerase chain reaction) amplification, sequencing and phylogenetic analysis were conducted to confirm the isolated species and genotypes of amoeba. According to morphological characterizations, 18.6% of specimens were identified as FLA, which in 71% were Acanthamoeba by PCR method. Molecular analysis revealed that 36.3%, 18.1% and 4.5% of Acanthamoeba specimens were identified as T3, T4 and T11 Acanthamoeba genotypes, respectively. Protacanthamoeba bohemica (27.2%) and Acanthamoeba sp. (4.5%) were found among the specimens. The results of osmo-tolerance and thermo-tolerance assays demonstrated that 50% of T3 and 25% of T4 genotypes of Acanthamoeba were highly pathogenic parasites. The molecular approach showed the presence of Naegleria lovaniensis (9%) in hot tub water of swimming pools. This study demonstrated that the swimming pools and hot tub water in Qazvin province were contaminated with Acanthamoeba and Naegleria species.

Molecular epidemiology of Blastocystis spp. in children referred to Qods hospital in northwest of Iran.

Blastocystis sp. is a polymorphic intestinal parasite in humans and animals. The parasite has a worldwide distribution, especially in developing countries with poor sanitation, exposure to animals, and improper disposal systems. The aim of this study was to identify the subtypes of Blastocystis sp. among children of Qazvin, northwest Iran. Totally, 864 stool samples were collected from the children referred to Qods hospital in Qazvin, Iran. Fecal specimens were investigated by formalin-ethyl acetate concentration method and trichrome staining as well as cultivation of all samples in clotted fetal bovine medium. DNA extraction of culture-positive specimens and PCR amplification of 18S ribosomal RNA gene region was performed. The sequences detected were compared with reference genes in the GenBank, and the sequences further deposited in the GenBank database. Data analysis was performed by Chi square test while a p value of < 0.05 was considered as significant. Of 864 isolates, 4.1% (36/864) were positive for Blastocystis sp. with infection rate insignificantly higher among the females than males. The highest infection rate was estimated at 6.8% in 6-9 years old age group with abdominal pain as the most common (33%) gastrointestinal sign. No statistically significant difference was found between the variables and Blastocystis infection. Molecular analysis clarified the presence of three subtypes of Blastocystis including ST1 (56%), ST2 (28%), and ST3 (16%) of among specimens with ST1 as the predominant subtype. A significant association between intestinal signs and the subtypes was not found. Considering ST1 as the predominant subtype, it seems that zoonotic transmission is a main route of human infections with Blastocystis sp. in the area studied.

Distribution and molecular analysis of Blastocystis subtypes from gastrointestinal symptomatic and asymptomatic patients in Iran.

INTRODUCTION: Blastocystis is a common intestinal parasite of human and animal hosts. The parasite has 17 subtypes, and among those at least nine subtypes (ST1-ST9) are found in human hosts. OBJECTIVE: The aim of the present study was to investigate the presence of different subtypes of Blastocystis spp. among the patients referred to Velayat hospital of Qazvin province, Iran. METHODS: Overall, 864 stool samples were examined by using formalin-ethyl acetate concentration method and Trichrome staining. All specimens were cultured in clotted fetal bovine medium. Later, DNA extraction and PCR amplification of 18S ribosomal RNA gene region was conducted and phylogenetic tree constructed. RESULTS: The results revealed 7.9% (68/864) of the study population were infected with Blastocystis. Intestinal symptoms were observed in 61% (36/59) of individuals positive for Blastocystis, with abdominal pain in 58% (21/36) of cases which was more frequent than other intestinal signs. No significant relationship was observed among the study variables. By molecular and phylogenetic analysis, three subtypes ST1 (45%), ST2 (30%) and ST3 (23%) of parasite were identified. CONCLUSION: This study showed ST1 subtype was the predominant subtype among the positive specimens, meanwhile the highest haplotype and nucleotide diversity were clarified in ST3 subtype.

Molecular Characterization of Fasciola and Dicrocoelium Species Isolated from Ruminant Livestock in Qazvin, Iran.

INTRODUCTION: Fascioliasis and dicrocoeliasis are the most frequent zoonotic diseases with increasing human health problems in different parts of Iran. Two species, Fasciola hepatica (F. hepatica) and Fasciola gigantica (F. gigantica), are spread in the country. Molecular approaches have a decisive role in identifying both the species. The aim of this study was to detect Fasciola spp. and Dicrocoelium spp. by amplifying the ITS-2 and 28S rDNA gene sequence. METHODS: Overall, 30 infected liver samples were collected from the livestock of Qazvin, Iran. The adult flukes were collected from different livestock. DNA extraction and PCR amplification of ribosomal RNA gene region (ITS2) and 28S rDNA gene fragment were conducted and a phylogenetic tree was constructed. RESULT: All the isolates obtained from the cattle (No: 7) and 82.6% (No: 19) of sheep isolates were infected with F. hepatica species, whereas 17.4% (No: 4) of sheep isolates were infected with F. gigantica. It was also shown that F. hepatica was the predominant species of Fasciola present in the region. All the specimens were infected with Dicrocoelium dendriticum (D. dendriticum). CONCLUSION: Both the species of Fasciola were found in Qazvin. D. dendriticum was the sole infecting species of the Dicrocoelium genus in the livestock of the city of Qazvin. Further research studies are needed to determine the intermediate host of the parasites in the region.

Molecular characterization of Hymenolepis nana based on nuclear rDNA ITS2 gene marker.

INTRODUCTION: Hymenolepis nana is a zoonotic tapeworm with widespread distribution. The goal of the present study was to identify the parasite in the specimens collected from NorthWestern regions of Iran using PCR-sequencing method. METHODS: A total of 1521 stool samples were collected from the study individuals. Initially, the identification of hymenolepis nana was confirmed by parasitological method including direct wet-mount and formalin-ethyl acetate concentration methods. Afterward, PCR-sequencing analysis of ribosomal ITS2 fragment was targeted to investigate the molecular identification of the parasite. RESULTS: Overall, 0.65% (10/1521) of the isolates were contaminated with H. nana in formalin-ethyl acetate concentration. All ten isolates were succefully amplified by PCR and further sequenced. The determined sequences were deposited in GenBank under the accession numbers MH337810 -MH337819. CONCLUSION: Our results clarified the presence of H. nana among the patients in the study areas. In addition, the molecular technique could be accessible when the human eggs are the only sources available to identify and diagnose the parasite.

Prevalence of Parasitic Intestinal Infections Among Food Handlers in Qazvin, Iran

Objective: Food handlers play an important role in the production and distribution of parasitic contaminations in these societies. The aim of this study was to assess the prevalence of intestinal parasitic infections among the food handlers in Qazvin, Iran. Methods: Totally, 1530 stool samples were collected from food handlers in Qazvin, Iran. Direct feces examination and formalinethyl acetate concentration method followed by Trichrome staining were performed to detect the parasites. Results: The prevalence of intestinal parasites was 82 (5.4%) among food handlers. The species of protozoan parasites found were Blastocystis hominis, Giardia lamblia, Entamoeba coli, and Endolimax nana. The outbreaks of Blastocystis hominis (2.9%), and Endolimax nana (0.3%) were the most and the least protozoan infections observed among the specimens, respectively. Also, there was no helminthic infection detected among the samples. The prevalence of intestinal parasites among the food handlers, based on variables including sex, age, job, and education, did not differ, whereas the prevalence of intestinal symptoms was shown to be higher in infected food handlers (p<0.05). Conclusions: Despite a decrease in the incidence of intestinal parasites, the protozoan infections are still among the health concerns in these areas. Therefore, continuous monitoring and training of food handlers by local health authorities should be performed on a regular basis.

Prevalence of intestinal parasites in food handlers of the city of Saqqez in 2016.

This study was aimed at determining the prevalence of parasitic infections among the food handlers of Saqqez County in 2016. Stool samples collected from 1530 food handlers. All samples were examined by both direct technique and sedimentation technique using formalin-ethyl acetate for detecting the intestinal parasites. Of total participants, 1462 (95.6%) were male and 62 (4.4%) female. Intestinal parasitic infections were found in 92 (6.01%) individuals. The highest infection rate was due to Giardia lamblia (4.1%) and the lowest associated with Endolimax nana, Iodamoeba butschlii, and Hymenolepis nana (0.1%). No significant correlation between the intestinal parasitic infections and age, gender, education level, gastrointestinal signs and symptoms, and the type of profession, was established. The results of the present study revealed that only a small number of food handlers in Saqqez County were infected with intestinal parasites with G. lamblia as the most common cause of infection. More strict health control for those working in food industries is of vital importance and continuous education to improve the level of general health in these people is undoubtedly considered as a key determinant towards lower number of parasitic infections.

A serologic study on Toxoplasma gondii infection in slaughtered sheep and goats in Qazvin Province, Iran.

BACKGROUND: Toxoplasma gondii is a common protozoan parasite among all mammals, in particular small ruminants, worldwide. Traditional husbandry can be a major risk factor for infection of sheep and goats with this parasite. OBJECTIVES: The present study aimed to determine the current status of the prevalence for T. gondii in livestock of Qazvin Province. METHODS: In this cross-sectional study, the sera of 455 sheep and 375 goats were examined to detect anti-Toxoplasma IgG antibodies by using in-house indirect ELISA. RESULTS: Overall, 33.62% (153/455) of sheep and 36.41% (130/375) of goats were positive for anti-Toxoplasma IgG antibodies with no statistically significant difference. The prevalence rate of T. gondii among the sheep of Qazvin County was significantly higher than in Abyek and Abhar counties (p < 0.001). CONCLUSIONS: The results of the present study indicate that the prevalence of T. gondii in sheep and goats of the study area is high. Therefore, the meat of the animals reared in this area can be a potential source of human infections by this parasite.

Infection with Hymenolepis diminuta (Rodolphi, 1819) in a Child from North of Iran: Case Report.

We report a human case of Hymenolepis diminuta infection in Guilan Province, northern part of Iran in 2017. The patient was a 15-month-old boy with gastrointestinal symptoms. In stool examination, eggs of H. diminuta was found based on morphological characteristic. The infant was successfully treated with a single oral dose of praziquantel, and then completely recovered. For the first time, we report human infection with this species in north of Iran.

Investigation of Giardia intestinalis Genotypes among the Food Handlers of Qazvin, Iran.

BACKGROUND: We aimed to investigate the genotypes of Giardia intestinalis among the food handlers in Qazvin, Iran. METHODS: Overall, 1530 stool specimens were collected from the food handlers who visited Shahid Bolandian Health Center, Qazvin, Iran during 2016. Specimens were evaluated by microscopic and concentration methods. Twenty specimens with appropriate number of giardia cysts were selected followed by DNA extraction. Determination of giardia genotypes was achieved through PCR and sequencing the glutamate dehydrogenase gene. The phylogenetic tree was drawn using the MEGA7 software. Finally, the data were analyzed statistically with a P-value<0.05 was considered as significant. RESULTS: Twenty stool samples (1.3%) were positive for Giardia cyst. All positive specimens were obtained from male participants with abdominal cramp being their most common symptoms. The mean age for infected individuals was 32 yr. Molecular characterization was successfully performed for 17 isolates and two genotypes A (AII, 65%) and B (BIII, 35%) were identified. CONCLUSION: The most prevalent giardia genotypes among the food handlers in Qazvin were A (AII) and B (BIII) genotypes with A (AII) genotype as the dominant one in the region. Considering the direct association between the food handlers and public health as well as the impact of geographical and host conditions on dispersion and pathogenicity of various genotypes and their zoonotic aspects, further investigations are necessary.

Prevalence of Intestinal Parasites among Rural Residents of Takestan in North-West of Iran.

BACKGROUND: Intestinal parasites are one of the health challenges in developing countries. Decreasing the prevalence of intestinal parasitic infections (IPIs) is one of the main aims of health services in these countries. This study was designed to determine the current status of IPIs in rural residents of Takestan a town located in North West of Iran. METHODS: A total of 2280 rural residents of Takestan were randomly selected. Data were collected through questionnaire by interviews and laboratory findings obtained by microscopic examination of stool sample including wet smear and formalin ethyl-acetate concentration. A P<0.05 was considered significant, statistically. RESULTS: In total, 8.7% (199/2280) of participants were positive for at least one intestinal parasite. The prevalence of polyparasitism was 0.7% in study population. Hymenolepis nana was the only helminthic infection which was detected (1/2280). Blastocystis, Entamoeba coli, and Giardia lamblia were the most common IPIs with prevalence of 3.6%, 2.9%, and 1.6%, respectively. Statistically, the prevalence of IPIs showed significant differences among villages (P<0.01) and age groups (P<0.001), and also habit of eating raw vegetables (P<0.005), whereas, the difference was insignificant in terms of sex, education level, and occupation. CONCLUSION: The prevalence of IPIs in rural residents of the study area is considerably low and this reduction was very impressive about helminthic infections.

Evaluating the effect of Myrtus communis on programmed cell death in hydatid cyst protoscolices.

OBJECTIVE: To evaluate the possible involvement of programmed cell death strategy in hydatid cyst protoscolices following treatment with Myrtus communis (M. communis) as an herbal medicine. METHODS: Protoscolices were aseptically collected from sheep liver hydatid cysts. Evaluating the effect of M. communis extract on programmed cell death and increased activity of caspases 3, 8, and 9 in hydatid cyst protoscolices was conducted by treating the protoscolices with different concentration (5, 50, and 100 mg/mL) of M. communis extract at 37 °C and 5% CO2 for 4 h by using the Bradford test and ELISA commercial kits. RESULTS: The extract of M. communis at all concentrations led to initiation of programmed cell death in protoscolices and this effect, was only significant at 50 and 100 mg/mL concentrations, compared to the negative control (P < 0.05). Also, the activity of caspases 3, 8, and 9 in hydatid cyst protoscolices, was shown that the extract at all 3 concentrations could only increase the activity of caspases 3 and 9. Moreover, a significant increase in the activity of caspase 3 was only observed at concentrations 50 and 100 mg/mL by 37.00% and 66.19% while a significant increase in the activity of caspase 9 at the same concentrations was observed by 20.89% and 63.67%, respectively (P < 0.05). CONCLUSIONS: The extract of M. communis at different concentrations could increase the activity of caspases 3 and 9 and caused programmed cell death in hydatid cyst protoscolices however, this effect was significant at high concentrations of the extract.