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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.

Sarcocystis species is a genus of cyst-forming parasites infecting both humans and animals globally. Some of these species cause clinical and subclinical diseases in the host and may lead to economic losses. This study was carried out to identify the distribution patterns of Sarcocystis spp. in slaughtered sheep based on the digestion method and PCR-RFLP in Isfahan, the center of Iran. In total, 150 fresh muscle samples (30 hearts, 60 esophagi, and 60 diaphragms) were investigated by naked eye observation and then scrutinized based on the digestion method. To this end, pepsin and HCl were used to observe the Sarcocystis parasite via a light microscope. The PCR was carried out to intensify a fragment of the 18S rRNA gene. Afterward, the PCR products were exposed to digestion by endonuclease TaqI, HindII, EcoRI, and AvaI. Consequently, the results of RFLP were confirmed by sequencing, and the phylogenetic placement of all species was analyzed. Through the examination by the naked eye, 5/150 (3.33%) macroscopic cysts were found in the samples. With the tissue digestion and microscopic examination, 116 (77.33%) samples were positive for Sarcocystis spp.; however, 125 (83.33%) samples were positive with PCR. Moreover, the results of sequence analysis on macrocysts and microcysts showed that 4% and 96% of the species belonged to S. gigantea and S. tenella, respectively. According to the results of the current study, sarcocystosis caused by S. tenella are highly prevalent among sheep in the Isfahan region. Due to the high prevalence of Sarcocystis infection in the world and Iran, the development of disease control and prevention policies in sheep would be essential, and changing attitudes in the way of keeping livestock from the traditional type to the industrial method is recommended in this regard.

Background: In a light microscope, image acquisition with different component depths is difficult, and there are various approaches for solving this problem. One of the common approaches is Camera Lucida (CL). This method has some disadvantages such as time-consuming, handed problems in painting, causing user boring, and produce gray scale output images. Aims and Objectives: In this study, we purposed a novel-combined hardware and software method. In this article, we try to present an automated method for our designed microscope. Materials and Methods: We have done a project with designed code number 377,694 to design and implement an upgraded light microscope. That project was about automatic movement of a stage with closed-looped control of a servomotor. Furthermore, automated camera catches images in predefined positions. That project has acceptable results in different parts, which encourage us to work on this study. This study help specialist have good fixative of all components in a sample. It is about trying to have useful Lucida Camera (drawing tube) in an automated scheme. Results: This method is an acceptable usual way for microscopic specialists, but with some disadvantages. It is time-consuming and boring that effect on the accuracy of results. Hence, how can be good if automated similar method could be implemented is exciting and affective. This studies idea comes from the basis of manual drawing tube (CL) method. In this experimental study, we have taken 400 handed an image of microorganisms. Captured images are from its whole body or various organs. They have been captured in different z-axis positions of stage, and hence components with different depths could be focused. Each patch checked for its edge strength to choose highest resolutions sub image and reconstruct focused image like a puzzle. This process has been continued for all areas to merge and complete reconstructed image as output. Conclusion: Comparing edge strength with other images and mean square error with manual focused on confirm our method with pleasure outcomes. Furthermore, independent focusing of an internal component in a sample body has been surveyed. It helps to have better resolution in internal selected component for more analysis and replace in its primitive image. This article presents efficient consequences with good accuracy and saving time in process period, which could be useful in different microscopes types and various samples type.

This study was designed to evaluate the gene expression level of IFN-γ in murine leukocytes treated by Toxoplasma gondii products in vivo using quantitative real time-PCR (Q-PCR) method. The mice considered as test groups were treated separately by Toxoplasma lysate product (TLP), excretory/secretory products (ESPs) obtained from cell free and cell culture media as well as active tachyzoites. The mice in the control group received phosphate buffered saline (PBS). The peritoneal leukocytes of the mice were harvested separately and their total RNA was extracted, converted to cDNA, and the gene expression level of IFN-γ was measured in all of the test groups relative to the control one. The results showed that the gene expression level of IFN-γ was decreased in the groups of the ESP from cell free medium (P=0.03), the TLP (P=0.02), and the active tachyzoite (P=0.16). Moreover, there was no significant difference statistically for the group of the ESP from cell culture medium (P=0.33). The findings revealed for the first time that the TLP and the ESP from cell free medium contain molecules that are capable of downregulating the expression of IFN-γ. Therefore, the question arises as to which molecules in these two products are involved in this process?

Background: Leishmaniasis infection threatens millions of people in under developing and developing countries. Treatment of this neglected disease is very complicated. Subjects and Methods: A novel series of antimony (V) complexes using bidentate ligands of hydroxypyranones and hydroxypyridinones have been designed and synthesized. For the synthesis of the complexes, SbCl5 in water was added to the solution of each ligand at 60°C and the pH of mixture was adjusted to 8 using aqueous NaOH. After 24 h stirring, extraction of produced compound into acetone gave the desired complex. The structure of complexes was achieved by using FTIR, 1HNMR, and electron spin ionization mass spectroscopic techniques. All compounds were evaluated for in vitro anti amastogote form of Leishmania major. Results and Conclusion: The most potent antimony complexes against amastigotes were 5b (after 48 and 72 h) and 5a (after 72 h) with IC50 values of 24.4, 16.3, and 30.1 µg/mL, respectively. Furthermore, antimony and iron complexes were used together for in vitro anti amastigote form of L. major activity. These compounds were toxic for macrophages and destroyed them.

Background: Today, leishmaniasis is a widespread, infectious parasitic disease caused by Leishmania spp. Natural-derived compounds are likely to provide a valuable source of new pharmaceuticals, and among them, quercetin derivatives may have antileishmanial effects. The antileishmanial activity of 3,5,7,3’,4’-pentahydroxyflavonol (quercetin) derivatives is partly attributed to the position and pKa of phenolic or catechol hydroxyl groups. Therefore, to optimize their leishmanicidal effect, the structural features of quercetin and its derivatives were improved by acylation or alkylation of hydroxyl groups and changing their pKa and consequently their activities. Materials and Methods: In this study, during a regioselective method, quercetin derivatives were synthesized. The structures of synthesized compounds were confirmed by mass, IR, 1H-, and 13C-NMR spectral data. The antileishmanial activities of compounds 1–6 were compared with glucantime as the standard drug against promastigotes of Leishmania major using standard cell-based leishmanicidal assay. Results: In this study, during a regioselective method, two 7-O-quercetin derivatives (5 and 6), and three quercetin acetate derivatives (2, 3, and 4) were synthesized. In detail, the IC50 values found against L. major were (1) 2.5 ± 0.92; (2) 2.85 ± 0.99; (3) 15.5 ± 1.95; (4) 13.5 ± 3.5; (5) 2.6 ± 0.57; and (6) 1.3 ± 0.35 μM while IC50 value of glucantime as the standard drug was 88.5 ± 9.47 μM. Conclusions: The present study showed an effective antileishmanial activity of quercetin semisynthetic compounds (1–6) against in vitro promastigotes of L. major. Among them, quercetin analogs with more lipophilic and iron-chelating activity showed more antiparasite activity.

Hydatid cyst is the larval stage of Echinococcus granulosus. In previous studies inhibitory effect of this parasite on cancer cell growth in culture medium has been shown. In this study effect of hydatid cyst antigens on tumor growth in experimental animals has been investigated. Two antigens of hydatid cyst including protoscolices excretory secretory antigen and hydatid fluid absorbed on alum as adjuvant were injected to two groups of C57/black mice as case groups. Control groups were injected with only saline and alum. All mice then were injected with melanoma cells. Both antigens reduced the tumor size in mice in case groups. The difference of tumor size in mice in case groups and control group was statistically significant. In conclusion, anti-tumor effect of hydatid cyst antigens may be related to antigenic similarities which exist between hydatid cyst and cancer cells.

Aim: The present study was aimed to evaluate E. granulosus genotypes isolated from goats using HRM analysis in Isfahan province. Background: Cystic echincoccosis, so-called hydatidosis, is widespread infection caused by the larval stage of Echinococcus granulosus. This is an important zoonotic disease worldwide, especially in the developing countries such as Iran. To date, molecular studies mainly based on the mitochondrial DNA sequences have identified distinct genotypes termed G1-G10 which can differ in some characteristics such as the growth and infectivity to different intermediate hosts or the survival rate in the definitive hosts that are important for the development of control strategies. Methods: From August to December 2014, 1341 goats were investigated and hydatid cysts were collected from the liver and lungs of 43 infected goats in Isfahan province abattoirs, Isfahan, Iran. Total genomic DNA was extracted from each sample, amplified for the presence of polymorphism of mitochondrial gene coding for cytochrome c oxidase subunit 1 (CO1), using high resolution melting curve (HRM) method. Results: the results of HRM analysis using the sequence of CO1 gene for 43 Echinococcus granulosus isolates from goats showed 31, 2 and 10 isolates were identified as G1, G2, and G3 genotypes, respectively. Conclusion: G1 is the predominant genotype in the isolated goat samples in Isfahan province, and the presence of G2 strain was reported for the first time in goat in Iran.

The avian hematozoa, with widespread prevalence worldwide, are able to infect a wide range of birds. There are various types of birds hosting hematozoa in Iran. The current study aimed at finding whether the avian hematozoa exists in Isfahan Province, central Iran. Since the study was the first of its kind in the province, use was made of “judgmental sampling” and, accordingly, Semirom, Chadegan, Fereydunshahr, Khomeyni Shahr, and Isfahan were selected as cities under study. In this study, birds belonging to five poultry families including ducks, turkeys, geese, hens/roosters, and pigeons—considered as native domestic birds—and a group of waterfowl migratory birds were studied. Blood samples were collected from 275 birds between May and June 2013. Then, a separate blood film was prepared for each sample and later, all the blood films were examined under a light microscope. The infection intensity was also determined for each of the infected birds. Two types of hematozoa, namely Aegyptianella (2.9 %) and Plasmodium (3.6 %) were found in the blood smear of the infected birds. The results revealed that the infection intensity was very low in all the infected cases. Further, the highest rate of the infected cases was related to the turkeys (12.2 %). In the geese, of course, no infection was observed. Fereydunshahr, with 14.2 %, was the city with the highest rate of infected cases. In Semirom, however, no infected cases were observed. In conclusion, more studies are necessary to clarify the status of various hematozoa species in other parts of Iran.

Toxoplasma gondii (T. gondii) is a well-known, commonly found pathogen by high proliferative activity in nucleated cells. This parasite has infected nearly one-third of the world’s population. Infection can result in chorioretinitis in immunocompetent hosts, encephalitis in HIV/AIDS positive patients and spontaneous abortion or neonatal death in fetuses infected with congenital toxoplasmosis. Development of vaccines for toxoplasmosis is of major importance for reducing routes of disease transmission among various host species in the world. Live attenuated vaccines resemble natural infectious agents very closely. Therefore, these vaccines are good enhancers of the immune system, often imparting lifelong immunity with only one or two doses. In this study, we review the current status of live attenuated vaccine development for Toxoplasma infection.

Background: Today, leishmaniasis is a widespread, infectious parasitic disease caused by Leishmania spp. Natural-derived compounds are likely to provide a valuable source of new pharmaceuticals, and among them, quercetin derivatives may have antileishmanial effects. The antileishmanial activity of 3,5,7,3',4'-pentahydroxyflavonol (quercetin) derivatives is partly attributed to the position and pKa of phenolic or catechol hydroxyl groups. Therefore, to optimize their leishmanicidal effect, the structural features of quercetin and its derivatives were improved by acylation or alkylation of hydroxyl groups and changing their pKa and consequently their activities. Materials and Methods: In this study, during a regioselective method, quercetin derivatives were synthesized. The structures of synthesized compounds were confirmed by mass, IR, 1H-, and 13C-NMR spectral data. The antileishmanial activities of compounds 1–6 were compared with glucantime as the standard drug against promastigotes of Leishmania major using standard cell-based leishmanicidal assay. Results: In this study, during a regioselective method, two 7-O-quercetin derivatives (5 and 6), and three quercetin acetate derivatives (2, 3, and 4) were synthesized. In detail, the IC50values found against L. major were (1) 2.5 ± 0.92; (2) 2.85 ± 0.99; (3) 15.5 ± 1.95; (4) 13.5 ± 3.5; (5) 2.6 ± 0.57; and (6) 1.3 ± 0.35 μM while IC50value of glucantime as the standard drug was 88.5 ± 9.47 μM. Conclusions: The present study showed an effective antileishmanial activity of quercetin semisynthetic compounds (1–6) against in vitro promastigotes of L. major. Among them, quercetin analogs with more lipophilic and iron-chelating activity showed more antiparasite activity.

Hydatid cyst caused by Echinococcus granulosus is one of the most important parasitic diseases around the world and many countries in Asia, including Iran, are involved with this infection. This disease can cause high mortality in humans as well as economic losses in livestock. To date, several molecular methods have been used to determine the genetic diversity of E. granulosus. So far, identification of E. granulosus using real-time PCR fluorescence-based quantitative assays has not been studied worldwide, also in Iran. Therefore, the aim of this study was to investigate the genetic diversity of E. granulosus from center of Iran using real-time PCR method. A total of 71 hydatid cysts were collected from infected sheep, goat, and cattle slaughtered in Isfahan, Iran during 2013. DNA was extracted from protoscolices and/or germinal layers from each individual cyst and used as template to amplify the mitochondrial cytochrome c oxidase subunit 1 gene (cox1) (420 bp). Five cattle isolates out of 71 isolates were sterile and excluded from further investigation. Overall, of 66 isolates, partial sequences of the cox1 gene of E. granulosus indicated the presence of genotypes G1 in 49 isolates (74.2%), G3 in 15 isolates (22.7%), and G6 in 2 isolates (3.0%) in infected intermediate hosts. Sixteen sequences of G1 genotype had microgenetic variants, and they were compared to the original sequence of cox1. However, isolates identified as G3 and G6 genotypes were completely consistent with original sequences. G1 genotype in livestock was the dominant genotype in Isfahan region, Iran.

Background: Given the fact that bruxism is a prevalent oral habit among children and a potential destructor of oral tissues, the present study aimed to investigate the relationship between intestinal parasitic infections and bruxism among kindergarten children. Methods: Questionnaires were administered among parents of kindergarten children in Isfahan to select 50 children identified by their parents to have the habit of bruxism and 50 without the habit as control group. Informed consent was obtained prior to the investigation. Parents were delivered sampling instruments with proper instructions to collect stool samples from both groups for parasitological tests. The diagnostic parasitological tests involved the direct stool smear, formol-ether concentration, and Scotch tape tests. Comparison for the frequency distribution of intestinal parasitic infections between the two groups was performed using the chi-square test (α = 0.05). Results: Parasitic infections were observed in 19 (11/50 cases and 8/50 controls) children. A statistically significant relationship was observed between infection with pathogenic parasites and bruxism (P < 0.05). Conclusion: Our findings suggest that pathogenic parasites may serve as the cause of initiation of bruxism habits among children.