Background:The number of valid of pathogen and non-pathogen species of Entamoeba has continuously increased in human and animals. This review is per-formed to provide an update list and some summarized information on Entamoeba species, which were identified up to the 2014.
Methods:We evaluated the Entamoeba genus with a broad systematic review of the literature, books and electronic databases until February 2014. The synonyms, hosts, pathogenicity and geographical distribution of valid species were considered and recorded. Repeated and unrelated cases were excluded.
Results:Totally 51 defined species of Entamoeba were found and arranged by the number of nuclei in mature cyst according to Levin's grouping. Seven of these spe-cies within the 4 nucleate mature cysts group and 1 species with one nucleate ma-ture cyst are pathogen. E. histolytica, E. invadence, E. rananrum and E. anatis causes lethal infection in human, reptiles, amphibians and brides respectively, four species causes non-lethal mild dysentery. The other species were non-pathogen and are important to differential diagnosis of amoebiasis.
Conclusion:There are some unknown true species of Entamoeba that available information on the morphology, hosts, pathogenicity and distribution of them are still very limited and more considerable investigation will be needed in order to clarify the status of them.

Backgrund:Free-living amoebae belonging to the genus Acanthamoeba have an environmental distribution. Amoebic keratitis due to these protozoan parasites continue to rise in Iran and worldwide. In Iran, there are various researches regard-ing both morphological and molecular identification of Acanthamoeba spp. in envi-ronmental and clinical samples. However, there is no thorough review about Acan-thamoeba genotypes and their distribution in environmental sources such as water, dust and biofilm in Iran. Besides, according to increasing cases of Amoebic kerati-tis in the region awareness regarding the pathogenic potential of these sight-threatening amoebae is of utmost importance.
 Methods:We conducted a thorough review based on the database sources such as MEDLINE, PubMed and Google scholar. No restrictions were placed on study date, study design or language of publication. We searched all valuable and relevant information considering the occurrence of the Acanthamoeba in both environmental and clinical samples.
Results:According to our thorough review Acanthamoeba belonging to T4 geno-type is the most prevalent type strain in environmental and clinical samples in sev-eral regions in Iran and worldwide, however, there are reports regarding Acan-thamoeba belonging to other genotypes such as T2, T3, T5, T6 and T11 and the mentioned point could leads us to more researches with the goal of presenting the real genotype dominance of Acanthamoeba and related disease in the country.
Conclusion:Overall, the present review will focus on present status of genotypes of Acanthamoeba in Iran during recent years.

Background:There are several methods, such as vaccination, to control visceral leishmaniasis. Although there is no efficient vaccine, it seem DNA vaccination with stimulates both cellular and humoral immunity apparently is the best way. The aim of this study was cloning and expression of LACK gene, a 36kD protein, as a can-didate protein for vaccination against Iranian L.infantum.
Methods:Iranian strain of L. infantum [MCAN/IR/07/Moheb-gh] was used as a template for PCR to amplify LACK gene. The LACK gene was cloned in pTZ57R/T vector and after confirmation it was digested by restriction enzymes (BamH1) and cloned in pcDNA3.1 expression vector. Recombinant plasmid was extracted and analyzed by sequencing, restriction digestion analysis and PCR reac-tion. The pc- LACK recombinant plasmid was purified from transformed E.coli (DH5α) and its expression was analyzed by SDS-PAGE and Western blot.
Results:The results of sequencing, restriction digestion analysis and PCR reaction revealed that LACK gene was cloned correctly in pcDNA3.1 vector and the results of SDS PAGE and Western blot emphasized that LACK protein of Iranian L. in-fantum is a well-expressed protein.
 Conclusion:We amplified, cloned and expressed Iranian L. infantum LACK genes successfully.

Background:This study was undertaken to evaluate the viability, infectivity and immunity of Toxoplasma gondii tachyzoites exposed to 2-(naphthalene-2-ylthio)-1H-indole.
 Methods:Tachyzoites of RH strain were incubated in various concentrations of 2- (naphthalene-2-ylthio)-1H-indole (25-800μM) for 1.5 hours. Then, they were stained by PI and analyzed by Fluorescence-activated cell sorting (FACS). To eval-uate the infectivity, the tachyzoites exposed to the different concentrations of the compound were inoculated to 10 BALB/c mice groups. For Control, parasites ex-posed to DMSO (0.2% v/v) were also intraperitoneally inoculated into two groups of mice. The immunity of the exposed tachyzoites was evaluated by inoculation of the naïve parasite to the survived mice.
Results:The LD50 of 2-(naphthalene-2-ylthio)-1H-indole was 57 μmol. The lon-gevity of mice was dose dependent. Five mice out of group 400μmol and 3 out of group 800μmol showed immunization to the parasite.
 Conclusion:Our findings demonstrated the toxoplasmocidal activity of the com-pound. The presence of a well-organized transporter mechanism for indole com-pounds within the parasite in conjunction with several effective mechanisms of these compounds on Toxoplasma viability would open a window for production of new drugs and vaccines.

Background:Acanthamoeba- bacteria interactions enable pathogenic bacteria to tolerate harsh conditions and lead to transmission to the susceptible host. The present study was aimed to address the presence of bacterial endosymbionts of Acanthamoeba isolated from recreational water sources of Tehran, Iran. To the best of our knowledge this is the first study regarding occurrence of bacteria in environmental canthamoeba spp. in Iran.
Methods:A total of 75 samples of recreational water sources were collected. Samples were cultured on non- nutrient agar 1.5% plates. Positive Acanthamoeba  spp. were axenically grown. DNA extraction and PCR reaction was performed using JDP1-2 primers. All positive samples of Acanthamoeba were examined for the presence of endosymbionts using staining and molecular methods. The PCR products were then sequenced in order to determine the genotypes of Acan-thamoeba and bacteria genera.
Results:Out of 75 samples, 16 (21.3%) plates were positive for Acanthamoeba according to the morphological criteria. Molecular analysis revealed that Acan-thamoeba belonged to T4 and T5 genotypes. Five isolates (35.7%) were positive for bacterial endosymbionts using staining method and PCR test. Sequencing of PCR products confirmed the presence of Pseudomonas aeruginosa and Agrobacterium tumefasiens.
Conclusion:The presence of Acanthamoeba bearing pathogenic endosymbionts in water sources leads us to public health issues including improved sanitation and decontamination measures in recreational water sources in order to prevent amoebae-related infection. To the best of our knowledge this is the first report regarding the isolation of A. tumefasiens from Acanthamoeba in Iran and world-wide. 

Background:Haemonchosis has a negative effect on the farming industry throughout the world, especially in the tropic and sub-tropic countries. The present study was carried out to differentiate Haemonchus species from its main hosts in Iran, including sheep, goat and camel.
 Methods:The identification took place based on the morphometrics of the spic-ules and molecular characters. Two hundred seventy adult male nematodes were collected from the abomasums of different ruminants (90 samples from each ani-mal) at the slaughterhouses from different localities in Iran. Samples were morpho-logically identified according to the spicules’ morphometric measurements. In the section on molecular study, 10 samples of each Haemonchus isolates were genetically examined. A simple PCR-restriction fragment length polymorphism (PCR-RFLP) assay of the second internal transcribed spacer of ribosomal DNA (ITS2-rDNA) were described to confirm the PCR results.
Results:PCR-RFLP profile obtained from the restriction enzyme HPa1 in H. con-tortus and H. longistipes indicated 1 (278 bp) and 2 (113 and 135 bp) different frag-ments, respectively. The morphological parameters clearly distinguish H. contortus from H. longistipes. Moreover, regarding the ITS2-rDNA, sequences of 295 bp and 314 bp were obtained from H. contortus and H. longistipes, respectively.
Conclusion:The genotypic results are in agreement with the phenotypic findings of both species.

Background:Haemonchosis has a negative effect on the farming industry throughout the world, especially in the tropic and sub-tropic countries. The present study was carried out to differentiate Haemonchus species from its main hosts in Iran, including sheep, goat and camel.
 Methods:The identification took place based on the morphometrics of the spic-ules and molecular characters. Two hundred seventy adult male nematodes were collected from the abomasums of different ruminants (90 samples from each ani-mal) at the slaughterhouses from different localities in Iran. Samples were morpho-logically identified according to the spicules’ morphometric measurements. In the section on molecular study, 10 samples of each Haemonchus isolates were genetically examined. A simple PCR-restriction fragment length polymorphism (PCR-RFLP) assay of the second internal transcribed spacer of ribosomal DNA (ITS2-rDNA) were described to confirm the PCR results.
Results:PCR-RFLP profile obtained from the restriction enzyme HPa1 in H. con-tortus and H. longistipes indicated 1 (278 bp) and 2 (113 and 135 bp) different frag-ments, respectively. The morphological parameters clearly distinguish H. contortus from H. longistipes. Moreover, regarding the ITS2-rDNA, sequences of 295 bp and 314 bp were obtained from H. contortus and H. longistipes, respectively.
Conclusion:High similarity of cloned PvMSP-142 kDa gene in comparison to reference sequence and other sequences could be beneficial as a remarkable mo-lecular marker for serological diagnostic.

Background:Acanthamoeba castellanii forms a resistant cyst that protects the para-site against the host’s immune response. Acanthamoeba Type-I metacaspase (Acmcp) is a caspase-like protein that has been found to be expressed during the encysta-tions. Dictyostelium discoideum is an organism closely related to Acanthamoeba useful for studying the molecular function of this protozoan caspase-like protein.
Methods:The full length of Acmcp and a mutated version of the same gene, which lacks the proline rich N-terminal region (Acmcp-dpr), were cloned into the pDneo2a-GFP vector separately. The pDneo2a-GFP-Acmcp and pDneo2a-GFP-Acmcp-dpr were electro-transfected into wild type D. discoideum cells to create cell lines that over-expressed Acmcp or Acmcp-dpr.
Results:Both cell lines that over-expressed Acmcp and Acmcp-dpr showed a sig-nificant increase in the fluid phase internalization and phagocytosis rate compared to the control cells. Additionally, the cells expressing the Acmcp-dpr mutant were unable to initiate early development and failed to aggregate or form fruiting bodies under starvation conditions, whereas Acmcp over-expressing cells showed the op-posite phenomena. Quantitative cell death analysis provided additional support for these findings.
 Conclusion:Acmcp is involved in the processes of endocytosis and phagocytosis. In addition, the proline rich region in Acmcp is important for cellular development in Dictyostelium. Given its important role in the development process, metacaspase protein is proposed as a candidate drug target against infections caused by A. castel-lanii.

Background:We have previously reported that a 31 kDa protein was screened from the excretory-secretory (ES) proteins of Tichinella spiralis muscle larvae (ML) by immunoproteomics using early infection sera, and the gene encoding a 31 kDa protein from T. spiralis was cloned and expressed in an E. coli expression system. In this study, the recombinant 31 kDa antigens were used for detection of anti- Trichinella antibodies in serum of experimentally infected mice by ELISA.
 Methods:Anti-Trichinella IgG antibodies in sera of mice infected with Trichinella were assayed by ELISA with recombinant 31 kDa antigens, and its sensitivity and specificity were compared with ELISA with ES antigen.
Results:The sensitivity and specificity of ELISA with recombinant antigens was 96.67% (29/30) and 96.87% (62/64), compared with 100% (30/30) and 98.44% (63/64) of ELISA with ES antigens was (P＞0.05). In heavily, moderately and lightly infected mice (500, 300 and 100 larvae/mouse), anti-Trichinella antibodies were firstly detected by ELISA with recombinant antigens at 8, 12 and 14 dpi, re-spectively; then increased rapidly with a detection rate of 100% respectively at 28, 22 and 30 dpi. While the antibodies were firstly detected by ELISA with ES anti-gens at 10, 8 and 10 dpi, respectively, the antibody positive rate reached 100% at 14, 12 and 22 dpi, respectively.
Conclusion:The recombinant 31 kDa antigens of T. spirali had a good sensitivity and specificity for detecting anti-Trichinella antibodies and might be the potential diagnostic antigen for trichinellosis.

Background:The aim of this study was to evaluate the effects of conjugated lino-leic acid (CLA) on apoptosis of tachyzoites of T. gondii, RH strain (type I) and the cyst-forming Tehran strain (type II) in vitro.
Methods:Toxoplasma strains were injected into the peritoneal cavity of BALB/c mice. The Tehran strain forms cysts in the brain of mice. Bradyzoites within the cysts are reactivated to proliferative tachyzoites, by dexamethasone. Tachyzoites were aspirated from the peritoneum of infected mice, and the percentage of viable parasites was estimated with trypan blue staining. Tachyzoites were inoculated into HeLa cells cultivated in DMEM medium. Different concentrations of CLA were evaluated on T. gondii in HeLa cells by the tetrazolium (MTT) colorimetric assay. Differentiation between apoptosis and cell death was determined by flow cytome-try using Annexin V and propidium iodide (PI) double staining. The statistical anal-ysis performed by GraphPad Prism version 6.00.
Results:CLA induces apoptosis in virulent (RH) and avirulent (Tehran) strains of T. gondii. The results of MTT indicated that CLA could decrease the proliferation of tachyzoites of both strains in HeLa cells.
Conclusion:Conjugated linoleic acid has anti-toxoplasmacidal activity on tachyzo-ites of T. gondii. Therefore, we recommended further studies on this component in order to achieve a new drug against the parasite.

Present paper is the second publication introducing the paleoparasitological findings from animal coprolites obtained from archeological site of Chehrabad salt mine in northwest-ern Iran. The current archeological site is located in northwest of Iran , dated to the Sas-sanian Era (4th/5th century CE).In the summer 2012 the carnivore coprolite was obtained within the layers in the mine and were thoroughly analyzed for parasites using TSP re-hydration technique. Eggs of Macracanthorhynchus hirudinaceuswere successfully retrieved from the examined coprolite and were confidently identified based on reliable references. Identifying of M.hirudinaceuseggs in paleofeces with clear appearance as demonstrated herein, is much due to appropriate preservation condition has been existed in the salt mine .The present finding could be regarded as the oldest acanthocephalan infection in Iran.

Background:Free-living amoebae such as Acanthamoeba species may act as carriers of Cryptosporidium and Toxoplasma oocysts, thus, may play an important role in the water-borne transmission of these parasites. In the present study, a loop mediated isothermal amplification (LAMP) method for detection of Toxoplasma and a PCR assay were developed for investigation of Acanthamoeba in environmental water samples.
Methods:A total of 34 samples were collected from the surface water in Guilan Province. Water samples were filtrated with membrane filters and followed by DNA extraction. PCR and LAMP methods used for detection of the protozoan parasites Acanthamoeba and Toxoplasma respectively.
Results: Totally 30 and 2 of 34 samples were positive for Acanthamoeba and Toxoplasma oocysts respectively. Two samples were positive for both investigated parasites.
Conclusion:The investigated water supplies, are contaminated by Toxoplasma and Acanthamoeba (oo)cystes. Acanthamoeba may play an important role in water-borne transmission of Toxoplasma in the study area. For the first time in Iran, protocol of LAMP method was used effectively for the detection of Toxoplasma in surface water samples in Iran.

Background: In leishmaniasis, some drugs prescribed for treatment have toxic ef-fects and there are reports about drug resistance in some countries. Due to this fact, using herbal drugs such as artemisinin with good efficacy and low toxic effect might be suitable.
Methods: We evaluated the apoptotic effect of artemisinin on Leishmania major in vitro and the antileishmanial activities of artemisinin on leishmaniasis in BALB/c mice and at the end INF-γ and IL-4 cytokines levels were detected by ELISA in spleen cell culture supernatants. During treatment the lesion size and survival rate were measured each four and ten days, respectively.
Results: Percentage of early and late apoptosis in promastigotes of control group and promastigotes treated with 10, 25, 50 and 100 μg/ml of artemisinin after 48 h were 0.13, 16.04, 41.23, 49.03 and 81.83, respectively. The IFN-γ in ointment treated group were higher than those of other groups (P<0.05). The in vivo results showed that ointment compounds healed the lesions more effectively rather than intraperito-neal injection method (P<0.05). The survival rate of mice 150 days after challenge in treated group with ointment of artemisinin was 66% while all mice in control groups were died.
Conclusion: All of in vitro results represented that this drug had antileishmanial ef-fects and these results were confirmed by evaluation effects in vivo condition of leishmaniasis. Interestingly, according to these results it can be concluded that this drug has antileishmanial effects in vitro and in vivo conditions. Artemisinin induces cytotoxic effect on L. major via apoptosis-related mechanism.

 Background: Parasitological investigations on paramphistomosis were carried out over a 12-month period in the southeast of Iran to determine the prevalence and intensity of this disease.
Methods: A total of 1000 cattle, Sistani breed (n= 450) and Brahman breed (n= 550) of all sex and age groups were inspected at random for the presence of paramphistomidae flukes in Zabol slaughterhouse from December 2012 to Octo-ber 2013.
Results: Paramphistomes were found in 369 of 1000 necropsied cows (36.9%; 95% CI: 30.1-41.9%), with significant higher prevalence of infection in Brahman breed than in Sistani breed (51% vs 19.3%). No significant correlation between prevalence, intensity of infection, sex and age of cattle was noted. Despite the dif-ference in the seasonal variations of prevalence, and the relation between the inten-sity of infection and season, these were not statistically significant. The mean inten-sity of infection in Brahman breed was higher (652.66 ± 281.5) than Sistani breed (123.32 ± 32.2). The identification of stained trematodes to the species revealed 40, 20, 20, 15 and 5% Gastrothylax crumenifer, Cotylophoron cotylophorom, Paramphistomum cervi, Carmyerius spatiosus, Explanatum explanatum, respectively.
Conclusion: The present results will contribute to our understanding of the epi-demiology of paramphistomumosis in southeastern Iran.

Background:Congenital toxoplasmosis is one cause of abortion. Infection can disrupt ovarian cycles and because toxoplasmosis is an infectious disease may have a similar effect on the ovaries. The purpose of this study was to investigate the pathological changes in the ovaries due to toxoplasmosis.
 Methods:Tachyzoites of Toxoplasma gondii were harvested from peritoneal fluid of mice, experimentally infected. Two females and one male mouse were housed per cage for mating in the overnight. The pregnant mice were divided into experi-mental and control groups. Experimental group were infected by parasite but the control group received the normal saline. The experimental and control mice were euthanized. Ovaries and uterine horns of animals were removed and prepared for light microscopy.
Results:Ovaries of infected pregnant mice presented gross morphological differ-ences compared to the control groups. In ovaries of experimental groups, changes of corpus luteum were observed. The comparison of experimental and control groups revealed that the number of primary follicles, secondary follicle, atretic pri-mary follicles and atretic secondary follicles had significant differences (P≤0.001).
Conclusion:Toxoplasma gondii alters ovarian follicular growth and development in mice. In addition, it alters number of different phases of follicles and corpus lu-teum in ovaries of mice.

Background: As a zoonotic pathogen, Encephalitozoon cuniculi is a cause of serious disease in animals and people. The present study was to evaluate the health status examination of this seropositive animal care worker in our pre-vious study.
Methods:Blood samples were taken from five workers. CIA test was ap-plied to detect antibodies against E. cuniculi in blood serum. The indirect immunofluorescence antibody test was used as confirmation test. Seroposi-tive worker had a complete medical examination.
Results:Only one worker was found to be seropositive according to the results of the serological test. Sera positive to E. cuniculi was confirmed with IFAT and spores were detected in the urine sample of the worker. The worker was treated with albendazole.
Conclusion:Rabbits should be examined routinely for the presence of anti-E. cuniculi antibody. People working with laboratory animal should avoid contact with urine and faeces of infected or pay attention to personal hy-giene.

Background: Toxocariasis is an important disease caused by the larvae of parasitic worms such as Toxocara canis and T. cati. Public parks can be the source of toxocariasis for small children. This survey was conducted to de-termine the prevalence of Toxocara spp. ova in parks of Mashhad and Khaf northeastern Iran.
Methods:In this descriptive cross-sectional study, performed in November 2011 to June 2012, overall, 340 soil samples were collected from 39 parks of Mashhad and 29 parks in Khaf city. Flotation method and direct smear were used, and the samples were evaluated using a light microscope. The results were analyzed using SPSS version 19 and Chi-square test.
Results:In the evaluation of 195 and 145 soil samples, 18 (9.2%) and 16 cases (11.3%) of contamination with Toxocara spp. eggs were detected, re-spectively.
Conclusion:Although the prevalence of Toxocara eggs in soil samples was low, parks can be a source of Toxocara infection of children in these areas.

Background: Encephalitozoon cuniculi is a microsporidian parasite commonly found in rabbits that can infect humans, causing encephalitozoonosis. Our objective in this study was to evaluate the seroprevalence of this parasite in rabbits and humans in China.
Methods: Overall, 300 serum samples each from clinically healthy rabbit and hu-man were collected from three regions of China (Sichuan Province, Chongqing Municipality and Jilin Province) from January to September 2013 and tested for anti- E. Cuniculi antibodies using an ELISA.
Results: An overall seroprevalence of E. cuniculi was recorded as 56/300 (18.76%) and 29/300 (9.76%) in rabbit and human sera, respectively. The seropositivity of rabbit samples collected from Jilin province was 41%, which was significantly higher (P<0.01) than Sichuan Province (9%) and Chongqing Municipality (6%). Three breeds of rabbit were used in the present study and antibody detection in Rex Rabbit was significantly (P<0.01) higher than Japanese White and New Zea-land Rabbit. In human, Jilin province was more prevalent (18%) followed by Si-chuan Province (6%) and Chongqing Municipality (5%).
Conclusions The E. cuniculi was present and widespread among healthy rabbits and humans in China.

Herein, a 28-year-old man with hoarseness, skin and oral lesions is presented. At the time of admission, the patient had an erythematous plaque on his chin near his lower lip and an erythematous-violaceous plaque on his palate near the open-ing of the pharynx and 20 kg weight lost in last one year. The biopsy of his skin lesions by hematoxylin and eosin staining revealed an infiltration of the dermis by lymphoplasma and histiocytic cells with a loose granuloma formation sugges-tive of leishmaniasis. Biopsy of mucosal lesions revealed Leishman bodies in dermis. PCR was performed on the specimens of skin, bone marrow, mucosa, and saliva, the results were positive. The pathogenic agent was identified as Leishmania major by the nested PCR. Serologic tests including direct agglutination test (DAT) and indirect immunofluorescence test (IFAT) were positive with high titers of anti-L. infantum antibodies (1:102400 versus 1:800, respectively), indicative of visceral involvement. The patient responded to a combination of miltefosine and meglumine antimoniate (Glucantime®). Visceral involvement due to L. major is rarely reported. To the best of our knowledge, probably hoarseness due to L. major has not been previously reported from Iran.

Coenurosis is a disease of the central nervous system in sheep, caused by Coenurus cerebralis, the larval stage of Multiceps multiceps, which inhabits the small intestine of Canidae. A case of regurgitations in a 2.5 month old lamb with acute coenurosis is being reported. The lamb was presented with a sudden onset of ataxia and re-gurgitations for 10 days. The post-mortem examination revealed 4 immature C. cerebralis cysts between 0.5 and 1.5 cm in diameter located in the brainstem and cerebellum, and histopathological examination revealed multifocal pyogranuloma-tous meningoencephalitis, so a diagnosis of acute coenurosis was established. Thus, acute coenurosis should be included in the differential diagnosis of regurgi-tations in lambs.