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Aims: In vitro cultivation of digenea would help the development of effective treatments and studies of the biology of the parasites. The goal of the present study was to optimize culture conditions for the maintenance liver fluke, Dicrocoelium dendriticum. Materials and Methods: Forty fresh D. dendriticum were collected from the sheep liver and washed three times with warm Roswell Park Memorial Institute (RPMI) 1640 Medium. The collected worms were transferred to 24-well Nunc-Immuno plates containing RPMI media supplemented with 50% of fetal bovine serum (FBS), 2% of sheep red blood cells (RBCs), 50 IU/ml of penicillin, and 50 mg/ml streptomycin. The mobility of the live/dead worms was observed by inverted microscope. The mean and median survival time was calculated by Kaplan–Meier model, and survival and hazard function graphs were also analyzed. Results: D. dendriticum was lived in vitro only for long periods of about 25 dyes. The 1st day of maintaining in culture media, one worm was dead and the number of dead worms was raised to 40 after 25 days of incubation. On the one hand, the mean survival time was 392 h with a confidence interval (CI) of 95% (384.8–400.03). On the other hand, the median survival time was 420 h with a CI of 95% (406.9–433.09). D. dendriticum was able to be alive in RPMI 1640 media for at least 25 days. Conclusion: RPMI 1640 supplemented with FBS, and RBCs can be used as short-term maintenance for the in vitro culture of D. dendriticum. The outcomes of the current study could be useful for many aspects of parasitological analysis.

Aims: Hydatidosis is an important zoonotic disease that is caused by a tiny tapeworm, namely Echinococcus granulosus. In this study, three polymerase chain reaction (PCR)-based methods, including, high resolution melting (HRM) analysis, DNA sequencing, and PCR-restriction fragment length polymorphism (RFLP) have been used for genotype the identification of E. granulosus isolates from dogs and camels in Zarinshahr and Najafabad, Isfahan province, Iran. Materials and Methods: A total of 200 adult worms of 40 dogs and 51 samples of camel hydatid cysts were examined. Molecular characterization of isolates was performed using HRM assay, sequencing of DNA, and digestion Rsa1 pattern coding for the mitochondrial cox1 gene. For analysis of the HRM melting curve, we used the Tm within the range of 77.50°C–79.23°C. Results: HRM analysis revealed that 72.5%, 15%, and 12.5% dog's genotypes and 41.17%, 21.56%, and 35.29% camel genotypes were G1, G3, and G6, respectively. PCR-RFLP analysis, spare parts 310 bp and 138 bp of cox1 that shows the G1 genotype in all of the isolates. Sequence analysis as well as HRM assay was confirmed genotypes of G1, G3, and G6 in camels and dogs. Based on three methods of the cox1 gene the dominant genotype was G1. Conclusion: The PCR-RFLP only identified the G1 genotype, whereas the HRM analysis, as well as DNA sequencing, were detected three genotypes G1, G3, G6, therefore, these two methods have enough accuracy for the determination of genotypes of E. granulosus. This information leads to a better understanding of the biological characteristics of E. granulosus genotypes in Iran and shows the camel as a source of human hydatidosis.

Aim: Fasciolosis and dicrocoeliosis are important parasitic diseases worldwide, causing significant financial losses due to decrease in production and viscera condemnation in animals. We performed the current research to assess the epidemiology of these infections and determine their significance from an economic perspective in Arak, Iran. Materials and Methods: In total, we evaluated 118,463 sheep, 207,652 goats, and 43,675 cattle through necropsic analysis at the slaughterhouses. The average weight of sheep, goat, and cattle liver was 1000, 900, and 5000 g, respectively. The average price of liver in the market was 8 USD/kg. Moreover, the elimination of fundamental nutrients and vitamins was evaluated in infected livers. The prevalence of fasciolosis and dicrocoeliosis was determined. Analysis of variance test was applied for the statistical analysis, and the significance level was

Recurrence of hydatidosis may results from cyst fluid spillage during surgery. To inactivate the cyst content, different scolicidal agents have been introduced. Nevertheless, novel and more effective treatments are needed due to the associated complications. In the current study, we examined the scolicidal effects of gold nanoparticles on Echinococcus granulosus protoscoleces. Hydatid cysts of sheep liver were collected in this study. The cyst fluid containing protoscoleces was aspirated aseptically. The protoscoleces were exposed to gold nanoparticles at different concentrations (250, 500, 1000, 2000, and 4000 μg/mL). Eosin staining method was applied to determine the viability of protoscoleces at 5, 10, 20, 30, and 60 min. Moreover, cytotoxicity analysis, scanning electron microscopy (SEM) of ultrastructural changes, DNA fragmentation, were performed, and protoscoleces were studied following treatment. The significant scolicidal effects of gold nanoparticles were observed at all concentrations compared to the control group. A total of 4000 μg/mL of gold nanoparticles could destroy 76% of protoscoleces in 60 min. Cytotoxic effects of gold nanoparticle on J774 macrophage cell line, in minimum and maximum concentration (500 and 4000 μg/mL), were 1 and 11%, respectively. After treatment of protoscoleces with gold nanoparticles in different times and concentrations, considerable alteration in size, ultrastructure changes in tegument and shape of sucker, as well as DNA fragmentation of parasite were seen. Based on the results, gold nanoparticles could exert scolicidal effects on hydatid cyst protoscoleces; therefore, they can be applied in hydatid cyst treatment.

The prevalence of bovine Sarcocystosis is high in the most regions of the world. It can be a human health problem due to consumption of raw or under cooked hamburgers or other bovine meat products. This study was carried out to investigate the prevalence and species identification of Sarcocystis among of hamburgers, using PCR–RFLP methods in Kashan, central Iran. Overall 200 raw industrial hamburgers samples with at least 60% meat were randomly collected from nine different brands in Kashan, central Iran. The genomic DNA was extracted and a PCR–RFLP method was used to amplify an approximately 900 bp fragment at the 18S rRNA(SSU) gene, restriction enzyme BclI was used for species identification. The results showed that 58 (29%) of 200 tested hamburger samples were infected to Sarcocystis spp. The prevalence rate was 31.25 and 26.9% in the hamburgers with 90 and 60–75% meat, respectively. According to PCR–RFLP analysis, 43 (74.1%) of the 58 isolates were Sarcocystis cruzi, 12 (20.7%) showed co-infection to S. cruzi and Sarcocystis hirsuta, 2 (3.5%) was mixed infected to S. cruzi and Sarcocystis hominis, 1 (1.7%) showed the pattern of mix infection to three species. This study revealed one-third of industrial hamburger were infected to S. cruzi or mixed infection of S. cruzi with other bovine sarcocytosis. To prevent cattle infection, the possible ingestion of the disposal sporocyst stage from dogs must be eliminated. Although in this study, the prevalence of S. hominis was low and cannot be considered as a major zoonosis, it should be recommended avoiding eating under cooked hamburger and other bovine meat products to prevent human infection.

Background and Aims: The aim of this study is to clarify nitric oxide (NO)-production by spleen and the importance of spleen in malaria infection in murine model. Materials and Methods: Thirty outbred NMRI female mice were divided into four groups, Group I: No intervention (Healthy control), Group II: With splenectomy (Healthy test), Group III: No intervention, Inoculation of contaminated blood (Infected control), Group IV: With splenectomy, inoculation of contaminated blood (Infected test). The Parasitemia was counted every other day through Giemsa stain examination of animal blood. The parasitemia and survival rates, hepatosplenomegaly and body weight were recorded. After terminal anesthesia, plasma and liver/spleen suspensions were assessed by the Griess micro assay for measurement of NO-levels. Results: At the end of the experiment (on day 16), the parasitemia was 26.99±0.46 % among the group of non-splenectomized animals (Group III) compared with 31.25±0.72% among the group of splenectomized animals (Group IV). The average parasitemia among the groups at the end of the experiment was statistically significant (Group III, Group IV: p= 0.0002). Survival rate was statistically significant (p

Polymerase chain reaction followed by high resolution melting (PCR-HRM) analysis is a simple, rapid and accurate method for molecular detection of various nematode species. The objective of the present study was, for the first time, to develop a PCR-HRM assay for the detection of various animal Trichostrongylus spp. A pair of primers targeting the ITS-2 rDNA region of the Trichostrongylus spp. was designed for the development of the HRM assay. DNA samples were extracted from 30 adult worms of Trichostrongylus spp., the ITS-2-rDNA region was amplified using PCR, and the resultant products were sequenced and characterized. Afterwards, the PCR-HRM analysis was conducted to detect and discriminate Trichostrongylus spp. Molecular sequence analysis revealed that 24, 4, and 1 of the samples were T. colubriformis, T. vitrinus and T. capricola, respectively. Results from PCR-HRM indicated that complete agreement was relatively found between speciation by HRM analysis and DNA sequencing for the detection of Trichostrongylus species. The PCR-HRM analysis method developed in the present study is fast and low-cost; the method can be comparable with other molecular detection techniques, representing a reliable tool for the identification of various species within the Trichostrongylus genus.

In the current era, cystic echinococcosis (CE), as larval stage of Echinococcus granulosus, is considered as a threat to human health. Scolicidal agents used in the surgery of cysts have different side effects. Therefore, the present study aimed to assess the effects of chitosan nanoparticles containing curcumin (Ch–Cu NPs) on the protoscolices of the hydatid cyst in vitro. Ch–Cu NPs were synthesized using a simple co-precipitation method and their structural and morphological properties were characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM), zeta analyzer, and Fourier transform infrared (FT-IR) spectroscopy. Then, the effects of different concentrations of Ch–Cu NPs (0.25, 0.05, 1, 2, and 4 mg/mL) on the fatality rate, and the length and width of protoscolices in different times (5, 10, 20, 30, and 60 min) were investigated. In addition, the SEM technique was used to evaluate the structure of the protoscolices after treatment. Based on the results, the presence of curcumin on the chitosan nanoparticles was confirmed by FT-IR analysis. Further, XRD analysis approved the crystal structure of chitosan NPs. Furthermore, the highest fatality rate was 68% in 4 mg/mL concentration of Ch–Cu NPs. The length and width of protoscolices decreased based on the high concentrations of Ch–Cu NPs, compared to the control group. Finally, Ch–Cu NPs expressed good scolicidal activities, which made them suitable to be considered as an anti-protoscolex agent.

The most important transmission path of intestinal parasites is the consumption of contaminated water and food. This survey was conducted to determine the prevalence of intestinal parasites among food handlers in Kashan, central Iran. This cross-sectional study was carried on 1018 food handlers who referred to the health centers in order to receive health certificate in 2017–2018. Stool samples of food handlers were collected and examined using direct and formalin-ethyl acetate methods. Results were recorded in information form along with some demographic data such as sex, age. The data were analyzed based on the tests of descriptive statistics by SPSS 16. Of the 1018 individuals examined 851 (83.6%) were men and 167 (16.4%) were female. Infection rate was 10.2% among the population and 9 types of parasites were diagnosed totally. Rate of infection to protozoan intestinal parasites were: Blastocystis sp. 7.17%, Giardia duodenalis 1.2%, Entamoeba coli 1.27%, Endolimax nana 1.08%, Dientamoeba fragilis 0.3%, Iodamoeba butschlii 0.3%, Entamoeba histolytica/E. dispar 0.2%, Chilomastix mesnili 0.1%. Hymenolepis nana (0.1%) was the only intestinal worm that observed. In this investigation, 8.9% of the individual were infected by one parasite, 1.08% by two and 0.2% by 3 or more parasites. This study showed that infection to intestinal helminthic is rare among food handlers in Kashan but the prevalence of protozoan intestinal parasites is high that similar to other regions of Iran. To reduce the rate of parasitic infections and inhibition of their transmission, continuing the process of promoting the health level is recommended.

Background: There are scant evidences between giardiasis and enterobiasis with human mal-absorption of micronutrient. Objectives: The aim of the present study was to found out the changes in the serum zinc, copper, magnesium and vitamin B12 levels in schoolchildren infected with Giardia intestinalis and Enterobius vermicularis. Patients and Methods: This case-control study was carried out among 359 children from aged 6 - 12 years old at 8 suburban public schools. Three stool examinations were done using standard techniques for identification parasitic infection. Fifty E. vermicularis and 37 G. lamblia infected individuals were enrolled as study groups. Serum levels of copper, zinc, magnesium were assessed by autoanalyzer and vitamin B12 levels were measured using gamma counter. ANOVA and paired t-test analyses were used to determine the association between infections and trace elements changes. Results: The prevalence of G. intestinalis and E. vermicularis infections were 10.3% and 13.9%, respectively. The serum zinc, copper and magnesium levels showed a significant decrease in individuals infected with G. intestinalis and E. vermicularis (P < 0.05). Mean values of Giardia positive and negative groups for copper 143.65 ± 16.51 and 176.26 ± 17.6 μg/dL, zinc 62.26 ± 16.06 and 80.66 ± 23.58 μg/dL, and magnesium 1.82 ± 0.23 and 2.01 ± 0.16 mg/dL, respectively. Mean values of Enterobius positive and negative groups for copper 145.55 ± 26.84 and 176.26 ± 17.6 μg/dL, zinc 72.7 ± 17.92 and 80.66 ± 23.58 μg/dL and magnesium 1.93 ± 0.11 and 2.01 ± 0.16 mg/dL, respectively. Conclusions: The results showed that giardiasis and enterobiasis decreased serum levels of copper, zinc and magnesium. Further studies are needed to clarify the actual mechanism governing the zinc, copper, magnesium and vitamin B12 giardiasis/enterobiasis interaction.

Trichomoniasis is a sexually transmitted disease (STD) caused by a tiny parasite called Trichomonas vaginalis. Metronidazole is used as routine treatment of disease. Some reports have confirmed the potential carcinogenic and teratogenic effects of this drug on fetus and indication of drug resistance. Verbascum thapsus belongs to the family of Scorphulariaceae. Its antiinflammatory properties, disinfectant and skin healing effects are well known. This plant has been used to treat diarrhea and genitourinary infection in traditional medicine. Effects of different concentrations of the Verbascum thapsus extract were tested on the growth and motility of T. vaginalis trophozoites. To evaluate the toxicity of extract, their effects on mice macrophages were measured by MTT([3-(4,5-dimethyl thiazolyl-2)- 2,5-diphenyle tetrazolium bromide ])assay. In this experimental study the effect of Verbascum thapsus ethanol extract on induction apoptosis in T. vaginalis was determined by Flow Cytometry. Results were analyzed by Flow Jo software and the degree of apoptosis was determined. Toxicity percentage of 25-800 μg/ml concentrations of Verbascum thapsus alcoholic extract for mice macrophages was observed between 0.17-0.25 after 12 hours and they were between 0.25-0.42 and0.45-0.95 after 24 and 48h respectively. IC50 (inhibitory concentration, 50%) of Verbascum thapsus ethanol extract and metronidazole after 24h was 39.17 and 0.0326 μg/ml respectively. Flow cytometry results showed the percent of apoptosis following treatment of trophozoites with different concentrations of Verbascum thapsus ethanol extract (25, 50,100,200,400 μg/ml), were 20.7, 37.04, 47.5, 62.72 and 86.35 respectively, while in control group was 2.9. According to this study, Verbascum thapsus extract induces programmed death in T. vaginalis. It is recommended that Verbascum thapsus extract can be considered as a suitable choice for Medical Studies.

Neospora caninum and Toxoplasma gondii are two closely related protozoan parasites that have been detected from various species of bird hosts. However, little is known about the prevalence of N. caninum and T. gondii in crows. Hence, we examined the molecular frequency of N. caninum and T. gondii in the brain samples of hooded crows (Corvus cornix) that collected from different public parks of Tehran, Iran by nested-PCR method. We used the primers targeting the Nc5 and GRA6 genes for detection of N. caninum and T. gondii, respectively. From a total of 55 brain samples, 5 (9.9%) and 9 (16.36%) samples were positive for N. caninum and T. gondii, respectively. Sequencing of a N. caninum isolate revealed 95%-100% identity with the deposited N. caninum in GenBank. Genotyping of T. gondii isolates by PCR-RFLP analysis of the GRA6 gene revealed type III genotype in 8 isolates. The results of this study indicate that hooded crows may have a putative role in transmission of N. caninum and T. gondii to canines and felines definitive hosts, respectively.

Little is known about latent infection and molecular characterisation of Neospora caninum in sheep (Ovis aries). In this study, 330 sheep samples (180 hearts and 150 brains) were analysed for N. caninum DNA by nested polymerase chain reaction (PCR) targeting the Nc-5 gene. Neospora caninum DNA was detected in 3.9% (13/330) of sheep samples. The parasite’s DNA was detected in 6.7% of heart samples (12/180) and 0.7% (1/150) of brain samples. No clinical signs were recorded from infected or uninfected animals. Sequencing of the genomic DNA revealed 96% – 99% similarity with each other and 95.15% – 100% similarity with N. caninum sequences deposited in GenBank. To our knowledge, this is the first report on the use of PCR to identify latent neosporosis in sheep in Iran. The results of this study have the potential to contribute to our understanding of the role of N. caninum-infected sheep in the epidemiology of neosporosis.

Background: Entamoeba histolytica/Entamoeba dispar and Enterobius vermicularis are the major health problems in the developing countries especially in Iran. The prevalence of infection is variable among different social groups in the world. Objectives: Since elderly and mentally retarded are high risk group, the present survey was carried out in order to determine the prevalence of intestinal parasites especially these parasites in elderly and mentally retarded residence in Golabchi Center, Kashan, Iran. Materials and Methods: In this Cross-sectional study a total of 243 stool samples and 279 Scotch tapes from elderly and mentally retarded people were collected. Intestinal parasitic infections especially E.histolytica/E.dispar was determined by Stool examination. Scotch tape was used for diagnosis of Enterobius vermicularis. The demographic data were recorded by questionnaire and were analyzed by SPSS and X2. Results: The overall infection rate of intestinal parasite was 78.7% (191 out of 243 subjects).The prevalence of E.histolytica/E.dispar and E.vermicularis in elderly were 16.8%, 25.5% and in mentally retarded 15%, 49.1% respectively. Prevalence of pathogenic parasites was: Taenia spp. 1.6%, Hymenolepis nana 0.8%, one case of Strongyloides stercoralis, Blastocystis hominis 33.3%, Giardia lamblia 4.5%, Dientamoeba fragilis 1.6%. The rate of infection in mentally retarded was higher than elderly (P

Background: Congenital toxoplasmosis (CT) is the result of maternal infection if acquired during gestation, an infection that most often is clinically unapparent. Objectives: This study was conducted to determine acute and chronic toxoplamosis in pregnant women and its complications in newborn. Patients and Methods: In this cohort study, 798 pregnant women were tested for Toxoplasma IgG/IgM by ELISA. PCR and IgM were performed on 4 newborns of acute and 5 chronic maternal infections; they were followed for 1 year. Data were analyzed by SPSS-11.5 and χ2 test. Results: Five (0.63%) and 341 (42.7%) were positive for acute and chronic infections. A 400 bp band was seen in 3 newborns of acute maternal infections. The incidence of neonatal toxoplasmosis was 3.7 per 1000. Only one neonate of the acute maternal toxoplasmosis had jaundice. Conclusions: The rate of mate

Neospora caninum is an intracellular protozoan parasite with a wide range of intermediate bird hosts. There is little information describing the prevalence and genetic characterization of N. caninum in bird hosts worldwide and in Iran. In this study, a total of 217 brain samples of house sparrow (Passer domesticus) were examined for N. caninum presence by nested polymerase chain reaction targeting the Nc-5 gene. N. caninum DNA was detected in 3.68% (8/217) of sparrows. Sequencing of the Nc5 genomic DNA revealed 97–99% of similarity with N. caninum sequences deposited in Genbank. To our knowledge, this study is the first molecular evidence of N. caninum DNA in bird hosts in Iran. The results of this study highlight the role of the house sparrow (Passer domesticus) in maintaining and spreading N. caninum infection to canines in the feral and domestic environment.

Toxoplasmosis is one of the most common parasitic diseases worldwide. It is estimated that approximately one-third of the world’s population is latently infected. Infection generally occurs via oral the route and maternal transmission. Damage of the central nervous system is one of the most serious consequences of congenital toxoplasmosis. Moreover, recent investigations proposed that acute and sub-acute congenital toxoplasmosis as well as latent toxoplasmosis during pregnancy; play various roles in the etiology of different neuropsychiatric disorders in mothers and their offspring. This paper reviews new findings about the role of latent toxoplasmosis in the etiology of various neuropsychiatric disorders in mothers and their offspring.

Objective To determine the prevalence and economic losses of Fasciola spp. isolates and costs incurred due to liver condemnation from ruminants in Kashan region, center of Iran from 2012 to 2013. Methods In this cross-sectional study, a total number of 251 325 slaughtered sheep, goats and cattle (88 939 sheep, 151 924 goats and 10 462 cattle) were examined via necropsy for fasciolasis in industrial Kashan abattoirs located in center of Iran. The average weight was calculated as 1 000 g for sheep, 900 g for goat and 5 000 g for cattle in this study region. Based on market value, the average price of a kilo gram of normal liver at Kashan city during the study period was 8 USD. The amount of main nutrients and vitamins elimination in liver contaminated to fascioliasis was determined. Results Overall 3.28% of the livers were found to be infected. For total number of sheep, goats and cattle slaughtered annually in region study, it was estimated that 7 505 livers were infected and total annual economic losses of fascioliasis of studied animals was 41 784 USD (based on market prices in study period). Of this, 23 360 USD, 30 240 USD and 15 400 USD were associated with the fascioliasis of sheep, goats and cattle, respectively. Conclusions This study indicated that the Fasciola sp. clearly remains prevalent and causes considerable economic losses in study region in sheep, goats and cattle and presumably, other areas and hosts of Iran, providing baseline data for the future monitoring of this potentially important parasitic infection in the country.

Background: Trichomoniasis is the most common sexually transmitted protozoan diseases in the worldwide. Metronidazole is the choice drug for trichomoniasis treatment, however, metronidazole resistant Trichomonas vaginalis (T.vaginalis) has been reported. Natural products are the source of most new drugs, and Zingiber officinale (Ginger) is widely used ingredient in the traditional medicine. Objective: The aim of the present study was to determine the effect of different concentrations of the ginger ethanol extract on the growth of T.vaginalis trophozoites in vitro. Materials and Methods: In this experimental study, 970 women who were attend in Kashan health centers were examined for T. vaginalis. Of them, 23 samples were infected with T.vaginalis. Three T. vaginalis isolates were cultured in a TYI-S-33 medium. The effect of ginger ethanol extracts and its toxicity in different concentrations (25, 50, 100, 200, 400, 800 µg/ml) on mouse macrophages were measured in triplicate exam by MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay. The effect of ginger on apoptosis induction was determined by Flow cytometry. Results: The IC50 of ginger and metronidazole were 93.8 and 0.0326 µg/ml, respectively. 12, 24 and 48 hr after adding different concentrations of extract on mouse macrophages, fatality rates in maximum dose (800 µg/ml) were 0.19, 0.26 and 0.31 respectively. Flow cytometry results showed the apoptosis rate following treatment with different concentrations of the extract after 48 hr were 17, 28.5, 42.1, 58.8, 76.3 and 100% respectively, while in the control group was 2.9%. Conclusion: Ginger ethanol extract induces programmed death in T. vaginalis. It is recommended that due to the known teratogenic effect of metronidazole, ginger can be considered as an alternative drug for metronidazole.