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BACKGROUND AND OBJECTIVES The current study was done to evaluate the relationship between T. gondii and obsessive-compulsive disorder (OCD) as well as prevalence rate of toxoplasmosis in treatment-resistant patients with OCD in comparison with treatment-sensitive patients with OCD. METHODS A total of 180 subjects were selected, including 90 patients with OCD and 90 control participants. Immunoglobulin G (IgG) and immunoglobulin M (IgM) antibodies against T. gondii were measured by the enzyme-linked immunosorbent assay (ELISA) technique. Detection of acute and chronic toxoplasmosis was carried out using the ELISA IgG avidity. In addition, the presence of this parasite in blood was detected using the nested-polymerase chain reaction (PCR) method. RESULTS Regarding T. gondii IgG antibodies 42 (46.7%) and 17 (18.9%) cases were detected in the patients and controls, respectively (P = 0.000). Also, 16 (17.8%) and 5 (5.6%) cases were positive for B1 gene in patients and controls, respectively (P = 0.018). The antibodies were found to be related to risk of OCD [OR (95% CI) = 3.71 (1.88-7.30); P

This study aimed to screen the natural infection rate of Leishmania major in Phlebotomus papatasi and Phlebotomus alexandri in two counties (Mehran and Dehloran) of Ilam province as cutaneous leishmaniasis endemic areas in the west of Iran. Furthermore, the genetic diversity of parasite species that are isolated from vectors, was investigated. Sandflies were collected by sticky traps from May 2018 to October 2018. Afterward, specimens were prepared for species identification by morphological features. DNA was extracted from female sandflies, and minicircle kDNA was used to identify Leishmania isolates through nested-PCR, followed by genetic diversity between Leishmania isolates was investigated by sequence analysis of the amplified minicircle kDNA. Natural infection of the L. major was shown in all positive specimens using nested-PCR. Analysis of data from 14 isolates displayed a high level of genetic diversity in L. major. In the phylogenetic trees, all of the L. major isolates occurred in six clusters. Clusters I, II, III, and VI contained isolated strains from P. papatasi. While clusters IV and V contained isolated strains from P. alexandri. Genetic diversity of L. major isolated from vectors was investigated in western Iran for the first time. According to the results of this study, probably “various clones of L. major populations are distributed in the study area.

Cutaneous leishmaniasis is a crucial vector-borne disease caused by various species of Leishmania and is transmitted by several species of sandflies. The present study was conducted to describe sand fly fauna on vectors of leishmaniasis and performing molecular identification of Leishmania isolates from them on the Iran–Iraq border. Entomological surveys were done from May to October 2016–2018 in 2 counties (Mehran and Dehloran) of Ilam province, west of Iran. Sandflies were collected by 40 Sticky Traps at each station. Samples were mounted for species identification using morphological characters of the head and abdominal terminalia. DNA was extracted from Phlebotomus papatasi females, and Leishmania isolates were identified through PCR on minicircle kDNA, followed by sequencing. A total of 5592 sandflies including 2 genera of Phlebotomus and Sergentomyia comprising 8 species of sand flies were detected. Leishmania major infection was detected in 3.33% of 300 tested female sandflies. Phlebotomus papatasi was predominant in outdoor and indoor resting places. Phlebotomus papatasi was determined as dominant vector of Leishmania major infection in Mehran and Dehloran counties, West of Iran. It seems the composition of sandfly species in the study area is almost similar to the other parts of Iran. A detailed description of the epidemiology and ecology of Phlebotomine sand flies needs to be established to accomplish effective vector control programs.

Aims Toxoplasma gondii is an obligate intracellular, protozoan that causes a high incidence of serious zoonotic parasitic disease in humans. In the present study the immune-protective efficacy of a DNA vaccine encoding SAG1 in combination with a gene sequence encoding FliC of Salmonella typhimurium (Toll-like receptor 5 agonist) was evaluated against acute T. gondii infection in mice. Methods and results Ninety-nine female inbred BALB/c mice were divided into nine groups of 11 mice and were immunized intramuscularly three times at three-week intervals (days 0, 21 and 42) and challenged with virulent T. gondii RH strain 4 weeks later. The immunization of pVAX1-SAG1 administered with pVAX1-fliC in mice indicated specific humoral responses, with higher IgG antibody titers and a mixed IgG1/IgG2a response than in other groups (with a predominance of IgG2a over IgG2b and IgG1). Also, the cellular immune response elicited high levels of IFN-γ and IL-12 cytokines and low levels of IL-4 production compared to traditional adjuvants. Furthermore, the mice vaccinated with pVAX1-SAG1+pVAX1-fliC survived for slightly longer after the last immunization and challenge with the T. gondii. Conclusion This investigation indicated that cocktail DNA vaccine encoded SAG1 gene of T. gondii and FliC can protect against acute toxoplasmosis.

Background Blastocystis, a common intestinal protozoan of humans and animals, infected more than 1 billion people around the world. This enteric protozoan is frequently reported in both healthy individuals and patients with gastrointestinal (GI) symptoms. Methods Three hundred and forty-five fecal samples including 151 GI patients and 194 healthy individuals were examined by microscopy, culture and PCR-sequencing techniques to determine Blastocystis frequency and subtype (ST) variation. Results The occurrence of Blastocystis was detected 56 (16.2%) and 85 (24.6%) by microscopy, culture and PCR methods, respectively. Out of the 85 positive patients, 60 (70.6%) were asymptomatic and 25 (29.4%) were symptomatic. The results of 41 successfully sequenced isolates identified 8 (19.5%), 8 (19.5%), and 25 (61.0%) ST1, ST2, and ST3, respectively. Conclusion This study has found that Blastocystis was more common in healthy individuals than GI patients. Another finding was that no correlation was found between clinical symptoms and Blastocystis STs.

Cystic Echinococosis is one of the important parasitic diseases that is considered as a problem economics and health in many parts of the world. Many efforts have been performed for controlling the disease in the world. To reach a reliable vaccine against Cystic Echinococosis is one of the important duty of governments. Several antigen of hydatid cyst for vaccine candidate have been evaluated. In this study, P-29 antigen has been used for this purpose. E.g P29 antigen was cloned in Escherichia coli and transfected into the Chinese hamster ovary cell for antigen proliferation and used for vaccination in Balb/c mice. The recombinant antigen E.g-29 was shown using Western blot test. Two dilution of DNA vaccine (pCEgP-29) including 50 µg/100 µl and 100 µg/100 µl were prepared. Twenty four Balb/C male 6–8 week mouse were divided in 4 groups. The groups were included in 2 vaccination groups (pcEg.P29 50 µg/100 µl and 100 µg/100 µl dilution) as immunized groups and 2 groups of plasmid and PBS as control. The mice were injected intramuscularly 3 times with 2 weeks interval. After 3 weeks from last injection, all groups were challenged intraperitonealy with 2000 protoscolices. After 5 months, the mice were euthanized by ketamine/xylasine injection and number, size, and weight of cysts were recorded. Immunization rate was up to 93% in vaccinated group when compared with the control group. The results of this study showed that rEg.P29 could be considered as an effective vaccine for controlling of E. granulosus prevalence in intermediated host.

Introduction: Cystic Echinococcosis (CE) and toxocariasis caused by the larval stages of intestinal dog worms including Echinococcus granulosus and Toxocara spp. are among the most widespread zoonotic diseases. Methodology: Four hundred municipal waste collectors were serologically evaluated for CE and toxocariasis. To identify the seropositive cases of CE, an ELISA test was performed using native AgB. Toxocara IgG detection was carried out using ELISA DRG kit (USA), and the seropositive cases were then examined by a Western blot kit (LDBIO, France) to confirm the positive ELISA results. Results: 15 (3.7%) workers were seropositive for CE according to the ELISA. A significant relationship was observed between being seropositive and having contact with soil and dogs. No significant correlations were observed between education and the prevalence of these diseases. Of the 15 seropositive workers for CE, ten worked in district 5 of Ahvaz. Toxocara IgG was identified in 11 (2.7%) cases using the ELISA; however, none of them were confirmed by Western blotting. Conclusion: The 3.7% rate of seroprevalence for CE in asymptomatic municipal waste collectors living in urban regions of Ahvaz suggests a high rate. The higher rate of infection among workers in district 5 is likely associated with the presence of stray and owned dogs in that area of the city. A prolonged exposure to contaminated soil, the lack of awareness about the risk of diseases that can be transmitted through waste and the lack of general availability of suitable personal protective equipment for waste collectors might cause infectious diseases.

Microsporidia are obligate intracellular parasites that produce spores. The infections caused by these parasites are mostly considered to be opportunistic in immunodeficient patients. Because of the zoonotic nature of microsporidia as well as the increasing prevalence of immunodeficiency diseases, the aim of this study was to evaluate the molecular diagnosis of Enterocytozoon bieneusi (E. bieneusi) and Encephalitozoon spp. in exotic birds in southwestern Iran. Initially, 816 stool specimens were collected and stained by modified trichrome (Weber) staining. The slides were explored using light microscopy. In the next stage, the extracted DNA was amplified using a multiplex/nested PCR method. RFLP with the Mnl1 restriction enzyme was used to differentiate the Encephalitozoon species in the products of the molecular analysis. Out of 816 samples, 138 and 181 cases were found to be positive by the staining and the multiplex/nested-PCR methods, respectively. Of the 181 samples, 103 and 78 samples were positive for E. bieneusi and Encephalitozoon spp., respectively. The Encephalitozoon species were 17 E. cuniculi, 52 E. intestinalis and 9 E. hellem. Of 103 E. bieneusi samples, 57, 39, 2 and 5 cases were detected as genotypes D, M, E and L, respectively. The results showed a relatively high prevalence of microsporidia in exotic birds, and according to the results of the genotyping, these birds can be an important source of microsporidiosis. It is essential that high-risk individuals, including patients with immunodeficiency diseases, receive accurate and valid information about the risk of direct and indirect contact with infected exotic birds. Keywords: Enterocytozoon bieneusi, Encephalitozoon spp., Microsporidia, Exotic bird, PCR, Iran

This study aimed to determine the prevalence and species of Cryptosporidium among HIV/AIDS patients in southwest of Iran. Two hundred fifty faecal samples from HIV patients were examined for the presence of Cryptosporidium oocysts using a conventional coproscopic approach. Such oocysts were detected in 18 (7.2%) out of 250 faecal samples. Genomic DNAs from 250 samples were then subjected to a nested-PCR-RFLP technique targeting different loci of 18S rRNA gene for species identification. Out of 250 samples, 27 (10.8%) were positive for different Cryptosporidium spp; Restriction patterns resulting from the digestion of the nested amplicon with restriction endonucleases VspI and SspI showed that C. parvum (70.38%) was the most prevalent species, followed by C. hominis (25.92%) and C. meleagridis (3.7%), respectively. The mean CD4+ T-cell count was 215 cells/μL. There was a strong association between cryptosporidiosis and CD4+ T-cell count (P = 0.000) with the highest prevalence recorded among patients with CD4+ T-cell count

Objective Contamination of soil and water with infected oocysts have an important role in both animals and humans toxoplasmosis. Cats shed oocytes by their feces that can survive in environment such as moist soil, warm and humid weather for months and even years. The present study designated to molecular detection of Toxoplasma gondii oocysts in stool samples obtained from cats in Ahvaz, southwest of Iran. Methods Initially, 486 stool samples were randomly collected from cats. After sucrose flotation method, the DNA was extracted and PCR was carried out using the amplification of the B1 gene and repeat element sequence (RE). For strain typing purpose, the genetic marker of SAG2 was used in nested-PCR. To perform RFLP procedure, the products of nested-PCR were digested using Sau3aI and HhaI enzymes. Results A total 486 stool samples were examined to the amplification of the 194 bp fragment of B1 gene that 35 samples (7.2%) were positive. All positive samples were confirmed by using another PCR that was amplified the 130 bp repeat element sequence (RE). The identified genotypes were type III (32 cases), mix of type I and III (2 cases) and one of the samples was type I. Conclusion Our findings revealed a relatively high prevalence of T. gondii oocysts in stool samples obtained from cats. It is essential that the high-risk people receive the information about the risk of direct and indirect contact with the animals.

Microsporidia is often considered as an opportunistic infection in patients with impaired immune systems such as patients with acquired immune deficiency syndrome, transplant recipients, children and old people. Due to the ability of the parasite to transmit from animals to human as well as the increasing prevalence of parasitic infections and immune deficiency diseases; therefore, the aim of this study was to evaluate molecular diagnosis of microsporidia strains in slaughtered cows of southwest of Iran. Initially, 256 stool samples of cows were collected from 5 regions of Khuzestan province, southwest of Iran and stained by modified trichrome (weber). Then, the parasite spores were examined by optical microscope. Total DNA was extracted from samples using DNA extraction kit for stool (Bioneer) and evaluated by multiplex/nested PCR method. The products of nested-PCR were explored by RFLP method using restriction enzyme MnlI. For genotyping, positive samples of RFLP were sequenced. Of 256, 21 and 48 samples were found positive by the staining and nested PCR tests, respectively. Of 48, 36 samples were Enterocytozoon bieneusi with genotype frequency D (22), J (9) and M (5). Also 9 were detected as Encephalitozoon species among which 2 were E. cuniculi, 6 were E. intestinalis and 1 was E. hellem Eventually, 3 samples were found positive for both Enterocytozoon and Encephalitozoon. The results showed that cows can be a source of microsporidia infections. Due to the zoonotic importance of this parasite and its ability to transmit from animals to humans; the detection and species determination of the parasite seems essential. The highest risk of infection is for individuals with impaired immune systems.

Blastocystis hominis (B. hominis) is a protozoan zoonosis which clinical signs of infection with this parasite has been reported to be more severe in patients with weakened immune systems than healthy controls. So, the aim of the study was to evaluate genomic analysis of B. hominis isolates obtained from patients with HIV-positive using locus SSU-rDNA. At first, 268 stool samples were randomly collected from patients with HIV-positive referred to health centers of Khuzestan province, southwest of Iran. Formol-ether and direct smear techniques were used for the detection of parasitic agents. After extracting DNA, the samples were analyzed by the PCR method. Finally, the subtypes were determined by the sequencing and PCR methods. New samples were used for the preparation of positive control sample; they were cultured in coagulant-serum biphasic cultivation media. Of 268 stool samples, 33 (12.3%) cases were detected positive for B. hominis using Formol-Ether technique but 51 (19%) cases were positive using molecular method. The most common isolates were related to the subtype III with 29 positive cases (56.8%), then, genotype I with 11 (21.6%) cases, 6 cases (11.8%) with genotype II, 3 (5.9%) combined cases with genotypes I and III as well as 2 cases (3.9%) with genotype VI. There was a significant difference between two groups of HIV-positive patients (infected with the parasite and/or without the parasite) in the term of the mean of TCD4-positive cells. The results indicated a relatively high prevalence of B. hominis in HIV-positive patients as well as our findings may represent that the number reduction of TCD4-positive cells has an effective role in the increased risk of the parasitic infection in HIV-positive patients.

Introduction: The infections caused by Toxocara spp. are considered as one of the most important zoonotic diseases in the world, especially in developing countries. Human toxocariasis, particularly in children, is acquired by playing in public parks. Hence, the aim of the current study was to detect Toxocara spp. in the soils of public parks of the city of Ahvaz, southwest of Iran, using the PCR and loop-mediated isothermal amplification (LAMP) methods. Methods: In this descriptive cross-sectional study, 260 soil samples were randomly collected from the different public parks of the city of Ahvaz. After performing zinc sulfate (ZnSO4) flotation technique, the DNA samples were extracted from the isolated Toxocara spp. eggs. Lastly, the extracted DNA was used for PCR and LAMP-based molecular detection. Results: Out of 260 specimens, 57 (21.9%) samples were found positive for Toxocara spp., using the PCR method, out of that 38 (14.6%) samples were positive for T. canis and 19 (7.3%) samples were positive for T. cati. Also, out of 260 specimens, 81 (31.1%) cases were positive for Toxocara species, using the LAMP method, among them 51 (19.6%) samples were found positive for T. canis and 30 (11.5%) samples were positive for T. cati. Kappa (κ) coefficient between PCR and LAMP showed a strong agreement (0.766, P-value=0.002). Conclusion: The obtained data showed a relatively high outbreak of Toxocara spp. in the public parks’ soils of the city, using the PCR and LAMP methods. Since the parasite can cause human toxocariasis, particularly in children; thus, the health authorities of the city of Ahvaz and other similar cities, especially in developing countries, must pay more attention to the hygiene of the public parks’ soils.

Introduction: The human being is considered a natural host for Trichomonas vaginalis (T. vaginalis), which causes trichomoniasis, the most frequent non-viral sexually transmitted infectious disease in the world. The current study aimed to evaluate the prevalence and sequencing of T. vaginalis in women with vaginitis. Methods: In the current research, 514 vaginal discharge samples were obtained from women with vaginitis. The specimens were evaluated by the direct wet mount examination, Dorset culture medium, and PCR technique. Primers were designed for the detection of TVK3/TVK7, TVA5/TVA6 genes specific for the identification of T. vaginalis. The PCR-positive samples were sequenced and compared with the sequences registered in the GenBank database. Results: Among the collected samples, 30 (5.83%), 45 (8.75%), 90 (17.50%), and 62 (12.06%) cases were positive for T. vaginalis when assayed by the direct wet mount examination, Dorset culture medium, and PCR technique (TVK3/TVK7, TVA5/TVA6 genes), respectively. There was no significant relationship between trichomoniasis and demographic characteristics of women, such as age, occupational status, mode of delivery, number of deliveries, educational level, and contraceptive methods (p˃0.05). The range of vaginal pH was between 5-7 in women with vaginitis, and there was a significant statistical correlation between the pH values and the infection rate (p Conclusion: This study confirmed a relatively high prevalence of T. vaginalis in the southwestern region of Iran. According to our results, the PCR method, especially detecting TVK3/TVK7 genes, was more sensitive than the direct wet mount examination and Dorset culture medium methods.

To detection and genotype of Toxoplasma gondii isolated from soil in Ahvaz, southwest of Iran. Between August 2011 and May 2012 at different sites located in the area of the Ahvaz city south west Iran. A total of 200 soil samples were taken from different points of the region. Oocysts were recovered using the flotation method. Then, PCR reactions targeting the GRA6 gene were performed for specific T. gondii detection. The positive samples were studied by RFLP (random amplified fragment length polymorphism) using MseI enzymes to confirm the parasite linage. Toxoplasma DNA was found in 18 samples. Among them, 12 samples were successfully genotyped as GRA6 type III and 6 as GRA6 Type II. This is the first investigation detecting and genotyping T. gondii oocyst in environmental soil samples of Ahvaz, South west of Iran. The results of this study indicated that soil contaminated with T. gondii oocysts especially in public park may play a role in the epidemiology of human toxoplasmosis in southwest of Iran.

Objective: To evaluate Entamoeba spp. diagnosis in patients with inflammatory diarrhea by staining, copro-antigen ELISA and multiplex PCR methods. Methods: In this descriptive cross-sectional survey, 200 stool samples were randomly collected during 2015–2016. The stool samples were evaluated microscopically for the presence of the parasite using direct and formalin-ether concentration and trichrome staining methods. Then, the stool samples were examined by copro-antigen ELISA (Biomerica Company) and multiplex PCR methods. Results: Of 200 samples, 17, 29 and 23 cases were positive for Entamoeba species by the staining, copro-antigen ELISA and multiplex PCR methods, respectively. Of 23 positive samples in multiplex PCR test, 13 and 10 samples were positive for Entamoeba dispar (E. dispar) and Entamoeba histolytica (E. histolytica), respectively. Conclusions: Our finding indicated a relatively high prevalence of Entamoeba species in patients with inflammatory diarrhea in Ahvaz city. Due to the complications of E. histolytica/ dispar infection, the health authorities of the city must pay more attention to control and prevent the transmission of E. histolytica/dispar to individuals.

Objective: To evaluate microsporidia species in pigeons of Ahvaz city, southwest of Iran by the staining and multiplex/nested PCR methods. Methods: Initially, 131 stool specimens were randomly collected from pigeons. The specimens were stained by the modified trichrome (Weber) and were microscopically examined. The DNA was extracted by DNA stool extraction kit (Bioneer) and was examined by the multiplex/nested PCR method. For differentiating the species of Encephalitozoon, the products of PCR were explored by the restriction fragment length polymorphism (RFLP) using the Mnl1 restriction enzyme. Results: Of 131 specimens, 22 and 18 cases were positive by the staining and the multiplex/ nested-PCR methods, respectively. Of 18, 7 and 11 samples were detected as Enterocytozoon bieneusi and Encephalitozoon spp., respectively. In addition, 2 Encephalitozoon cuniculi and 9 Encephalitozoon intestinalis were Encephalitozoon species. Of 7 Enterocytozoon bieneusi samples, 5 and 2 cases were detected as genotype M and L, respectively. Also, all Encephalitozoon cuniculi samples were identified as genotype II. Conclusions: Our findings showed a relatively high prevalence of microsporidia at pigeons and these birds can be a significant and important source of microsporidiosis. It is essential that the high-risk individuals including patients with immunodeficient diseases should be receiving the accurate and valid information about the risk of direct and indirect contact with the infected pigeons.

Background: Accurate and effective diagnosis of Toxoplasma gondii (T. gondii) infection can make a significant contribution to the control and prevention of the infection, especially in high-risk individuals. Today, polymerase chain reaction (PCR) is frequently applied for the detection of the infection. Objectives: The aim of this study was to evaluate the frequency and genotyping of T. gondii in blood and CSF samples of children with immunodeficiency. Methods: In the current descriptive cross-sectional study, a total of 170 blood samples and 170 cerebrospinal fluid (CSF) samples were randomly taken from 170 children with immunodeficiency. Firstly, the samples were evaluated using the polymerase chain reaction (PCR) method by targeting the RE gene, and then, positive samples were examined by the nested-PCR method, based on the SAG2 genomic target. For genotyping, PCR-restriction fragment length polymorphism (RFLP) assay was by the Hha I and Sau3A I restriction endonucleases. Results: From 340 samples, 23 (seven blood samples and 16 CSF samples) were detected positive by the PCR method. Of 16 CSF samples, all cases were found positive by the nested-PCR method, while all blood samples were negative. After genotyping CSF samples, T. gondii type II was found in 15 samples and one sample was a mix of both types II and III. Conclusions: The findings indicated that the RE target can be considered as a sensitive method for the diagnosis of toxoplasmosis in the CSF samples, especially in children with immunodeficiency. In addition, this study showed that genotype II was the prevailing genotype in the patients.

Giardia duodenalis, one of the most common intestinal protozoa, infects a wide range of vertebrates, including humans and animals. In this study, 84 Giardia duodenalis positive stool samples were collected from 3580 patients attending the Imam Ali Hospital and two public health centers in Andimeshk County, southwestern Iran. Stool samples were examined initially by microscopy, and then G. duodenalis was confirmed by SSU rRNA gene and genotypes were determined by amplification of the gdh and β-giardin genes. The SSU rRNA, gdh, and β-giardin genes were successfully amplified in 89.3%, 58.3%, and 51.2% samples, respectively. Of the positive samples for gdh and β-giardin, 40 isolates were successfully sequenced. Twenty-three isolates belonged to assemblage A, sub-assemblage AII, and 17 belonged to assemblage B. Of the 24 successfully amplified asymptomatic cases, 12 belonged to assemblage A and 12 belonged to assemblage B. The current study found that 64.3% of the patients were asymptomatic. From an epidemiological point of view, the high percentage of asymptomatic patients is important because of their role in the transmission of Giardia. The predominant assemblage was assemblage A, sub-assemblage AII. In general, therefore, it seems that most infections are probably transmitted by anthroponotic pathways in the region.

Objective To investigate the prevalence of intestinal parasitic infections among health card applicants attending Ahvaz East Health Center during 2012–2013. Methods In this descriptive cross-sectional study, stool specimens were collected between 14 614 and 12 444 people referred to Ahvaz East Health Center respectively in 2012 and 2013. Then, stool specimens were examined microscopically for the presence of parasite eggs, cysts and trophozoites using direct and formalin-ether concentration methods. Results Among 14 614 applicants for health cards in 2012, 1 693 cases were diagnosed with parasitic infections: Giardia lamblia 1 445 (85.35%), Entamoeba histolytica/dispar 31 (1.83%), Oxyuris (pin worm) 29 (1.71%), Hymenolepis nana 60 (3.54%), and Entamoeba coli 128 (7.57%). Out of 12 444 applicants for health cards during 2013, 632 cases were diagnosed with parasitic infections: Giardia lamblia 510 (80.69%), Entamoeba histolytica/dispar 20 (3.17%), Oxyuris 19 (3.01%), Hymenolepis nana 46 (7.28%), and Entamoeba coli 33 (5.86%). In both years the most outbreaks have been reported in the spring and autumn, and the lowest outbreak has been reported during the winter. Conclusions The protozoa outbreak, such as Giardia, is more than multi-protozoa whose outbreak may be the result of parasite easy transferred by water and nutrients and the parasite cyst resistance under the worse conditions as well as its simple cell division. The outbreak percentage in 2012 is more than 2013 which may be the result of improving the individual health care level.

Background: Microsporidiosis is considered as an opportunistic infection in immunodeficient patients. Objectives: Due to the increasing prevalence of parasitic infections and immunodeficiency diseases as well as the transmission risk of microsporidia from animals to human, the aim of this study was to evaluate molecular diagnosis of Enterocytozoon bieneusi and Encephalitozoon spp. by the multiplex/nested polymerase chain reaction (PCR) and staining methods in wild rats of Ahvaz city, southwest of Iran. Methods: Initially, 160 stool samples were collected from wild rats in different parts of Ahvaz city. The samples were stained by the modified trichrome staining and explored microscopically. The DNA was extracted using the DNA stool kit and examined by multiplex/nested PCR. For differentiating the species of Encephalitozoon, the multiplex/nested PCR products were explored by the restriction fragment length polymorphism (RFLP) method using the restriction enzyme of Mnl1. Results: Of 160, 26 cases were suspected positive for microsporidia spore by the staining. Also, of 160, 18 cases were positive by the multiplex/nested PCR method, that 14 and 4 samples were detected as E. bieneusi and E. intestinalis, respectively. Of 14 E. bieneusi samples, 12 and 2 cases were detected as genotype D and M, respectively. Conclusions: The findings revealed a relatively high prevalence of microsporidia infection in wild rats of the city and these animals can be a source of microsporidiosis. Due to the zoonotic potential of the microorganisms, high - risk individuals should be receiving the information about the risk of direct and indirect contact with the infected animals. © 2018, Jundishapur Journal of Microbiology.

Background: Cryptosporidium is a protozoan parasite that effects rodents, dogs, calves, humans, and cats. Infection with this parasite is known as cryptosporidiosis. Cryptosporidium spp. may induce clinical or subclinical signs in infected hosts. In the life cycle of this parasite infected dogs freely living in urban and rural areas of Khuzestan province are the definitive hosts that should be considered as a real problem in public health for humans. Objectives: This study aimed at determining the frequency of cryptosporidiosis in dogs in southwest of Iran. Methods: Overall, 350 fresh fecal samples were collected from domestic dogs living in 43 villages, from June 2012 to September 2013. All samples were investigated by Sheather’s concentration method and fecal smears were stained with modified Ziehl-Neelsen followed by light microscope examination, and polymerase chain reaction (PCR). Results: The results revealed that frequency of Cryptosporidium infection was 8% and 12.3%, using direct smear and molecular method, respectively. Conclusions: The present findings indicated that domestic dog feces from southwest of Iranmay contain zoonotic parasites such as Cryptosporidium spp. andmay be a potential risk for humans and other animals, especially when they contaminate the environment. The role of dogs as source of human infection should be investigated by further studies.

Objective To test for immunoglobulin G (IgG) and immunoglobulin M (IgM) antibodies in butchers in the province of Khuzestan in Iran and compare them with a control group from the general population. Methods Blood samples were taken from 110 butchers in five cities of Khuzestan, southwest of Iran to test for the incidence of Toxoplasma . The participants were accepted and tested on the basis of age, work experience, engagement in animal husbandry, and positive or negative Toxoplasma IgM and IgG results. Results The results showed that the seroprevalence of Toxoplasma IgG was 41.8% (56 cases) among the group of butchers in Khuzestan and 28.8% in the control group. The seroprevalence of Toxoplasma IgM was 1 case (2.2%) among butchers, while none in the control group. Conclusions A comparison of results indicated that butchers in Khuzestan were at the risk of toxoplasmosis. The greatest frequency was 17.6% (20 cases) in the group with age ranging form 35 to 44 years. In terms of work experience, the greatest frequency was 16.5% (18 cases) among butchers having 9–17 years of work experience.

Microsporidia are often considered as an opportunistic infection in patients with impaired immune systems such as transplant recipients and patients with acquired immune deficiency syndrome (AIDS). Due to the increasing prevalence of parasitic infections and immunodeficiency diseases; the aim of the study is to evaluate molecular identification of Enterocytozoon bieneusi and Encephalitozoon spp. in immunodeficient patients in Ahvaz, southwest of Iran. At first, 310 stool samples were collected from patients with immunodeficiency. The specimens were stained by modified trichrome (weber) and were examined microscopically. The extracted DNA samples were evaluated by multiplex/nested PCR method. The products of multiplex/nested PCR were explored by RFLP method using the restriction enzyme of Mnl1. Of 310, 93 samples were suspected positive for microsporidia by the staining. Also, of 310, 88 samples were positive by the multiplex/nested-PCR test that 62 samples were positive for E. bieneusi as well as 26 were detected as Encephalitozoon species that including 3 E. cuniculi, 19 E. intestinalis and 4 E. hellem. Of 62 E. bieneusi, 45, 16 and 1 were detected as genotype D, M and WL11, respectively. Also, Of 3 E. cuniculi, 1 and 2 cases were identified as genotype I and II, respectively. All E. hellem samples were included genotype 1A. Our findings revealed a relatively high prevalence of microsporidia species in immunodeficient patients. The highest risk of this infection is at individuals with impaired immune systems that it can be life-threatening in people with immune system dysfunction. It is essential that the high-risk people should be receiving the information about the risk of direct contact with infected individuals and animals.

Background: Toxoplasma gondii is one of the most common causes of latent infections in humans worldwide. Detecting anti-Toxoplasma antibodies in serum using serological tests is a common method to diagnose toxoplasmosis. Objectives: In the present study, an indigenous ELISA kit was prepared using tachyzoites from the RH strain of T. gondii, and its sensitivity and specificity were compared with those of commercial kits. Methods: To produce antigens, 0.02 mL of locally isolated T. gondii RH strain parasites along with 109 tachyzoites were injected into the peritoneal cavities of 50 laboratory mice (BALB/C). Parasites were collected after 4 days. After filtering and washing, the concentration of protein in sonicated tachyzoites was calculated using the Lowry protein assay. The dilution of antigen, serum and alkaline phosphatase conjugate was assessed in designing an indigenous ELISA method; then ELISA was performed based on these dilutions, and its sensitivity was determined using 200 serum samples. In addition, the specificity of the assay was evaluated using 40 serum samples from patients with tuberculosis, leukemia or hydatid cyst. Results: Indigenous ELISA was used to examine 100 serum samples containing anti-T. gondii IgG, with a sensitivity of 98% (commercial kits: 100%). Another 100 serum samples containing anti-T. gondii IgM were also tested, with a sensitivity of 99% (commercial kits: 100%). When 40 serum samples from patients with leukemia, hydatid cyst or tuberculosis were examined using anti-T. gondii IgG, the specificity was 100%, identical to commercial kits. However, the specificity of a similar test with anti-T. gondii IgM was just 28.6% for serum samples from leukemia patients, 21.4% for hydatid cyst and 16.7% for tuberculosis. Conclusions: We found that purified locally isolated soluble crude antigens of the RH strain of T. gondii from the peritoneal cavity of mice may be one of the most promising antigens for detection of human toxoplasmosis in routine screening.

Toxoplasmosis is a common parasitic infection in the world. Since increased free radicals and oxidative stress are reported in many parasitic diseases the purpose of the present study was to evaluate the oxidative stress in acute and chronic toxoplasmosis. RH strains ofToxoplasmatachyzoites were used in the present study. Twenty-five female rats were infected with the parasite while 25 other rats were as the control group that received normal saline. Zero-, 5-, 7-, 10-, and 45-day postinfection (DPI) blood samples were taken. Some parameters related to oxidant and antioxidants such as antioxidant enzymes, malondialdehyde, and total antioxidant capacity were measured. On day 7 after infection, GPX activity and GSH level were significantly increased and in the mentioned day the amount of total antioxidant capacity was significantly reduced. In other cases, there were no significant differences between the groups in different days. Overall, based on the results it seems that, on day 7 after infection, in infected rats responses to oxidative stress were triggered and led to decrease of total antioxidant capacity. Furthermore, glutathione was increased to cope with stress. It seems that probably antioxidant defense system entered the infection to the chronic phase and changed the parasites stage.

Background: Toxoplasmosis is an opportunistic protozoan infection with a high prevalence in a broad range of hosts infecting up to one-third of the world human population. Toxoplasmosis leads to serious medical problems in immunocompromised individuals and fetuses and also induces abortion and mortality in domestic animals. Therefore, there is a huge demand for the development of an effective vaccine. Surface Antigen 1 (SAG1) is one of the important immunodominant surface antigens of Toxoplasma gondii, which interacts with host cells and primarily involved in adhesion, invasion and stimulation of host immune response. Surface antigen 1 is considered as the leading candidate for development of an effective vaccine against toxoplasmosis. Objectives: The purpose of this study was to clone the major surface antigen1 gene (SAG1) from the genotype 1 of T. gondii, RH strain into the eukaryotic expression vector pVAX1 in order to use for a DNA vaccine. Materials and Methods: Genomic DNA was extracted from tachyzoite of the parasite using the QIAamp DNA mini kit. After designing the specific primers, SAG1 gene was amplified by Polymerase Chain Reaction (PCR). The purified PCR products were then cloned into a pPrime plasmid vector. The aforementioned product was subcloned into the pVAX1 eukaryotic expression vector. The recombinant pVAX1-SAG1 was then transfected into Chinese Hamster Ovary (CHO) cells and expression of SAG1 antigen was evaluated using Reverse Transcriptase Polymerase Chain Reaction (RT-PCR), Immunofluorescence Assay (IFA) and Western Blotting (WB). Results: The cloning and subcloning products (pPrime-SAG1 and pVAX1-SAG1 plasmid vectors) of SAG1 gene were verified and confirmed by enzyme digestion and sequencing. A 30 kDa recombinant protein was expressed in CHO cells as shown by IFA and WB methods. Conclusions: The pVAX1 expression vector and CHO cells are a suitable system for high-level recombinant protein production for SAG1 gene from T. gondii parasites and are promising approaches for antigen preparation in vaccine development.

Background: Because of the economic importance of the Arab race horses and also the role of Toxoplasma gondii and Neospora spp. in abortion and reproductive failure of these animals, we decided to perform this study. Objectives: We designed this study to investigate the seroprevalence of anti-Toxoplasma gondii and anti-Neospora spp. antibodies in Arab horses from 12 cities of Khuzestan province in southwest of Iran. Materials and Methods: From October 2009 to March 2011, a total of 235 blood samples were collected from jugular veins of Arab horses of different ages and genders from 12 cities of Khuzestan province. All the sera were tested for anti- Toxoplasma antibodies using the modified agglutination test (MAT) and the existence of anti-Neospora antibodies were tested using N-MAT for Neospora spp. Results: According to the MAT results, antibodies to T. gondii were found in 114 (48.5%) of 235 sera with titers of 1:20 in 84, 1:40 in 19, 1:80 in four, 1:160 in four, and 1:320 in three horses. According to the N-MAT results, antibodies to Neospora spp. were found in 47 (20%) of 235 sera with titers of 1:40 in 39, 1:80 in five, and 1:160 in three horses. We did not observe any statistically significant differences regarding age groups and genders between seropositive and seronegative horses for Neospora spp. using chi-square (χ 2 ) test, but it seemed that anti- Toxoplasma antibodies were more prevalent in older horses (≥ 10 years old). Conclusions: The results indicated that Arab horses are exposed to these parasites in southwest of Iran. Further research is required to determine the genomic structures of these parasites in Arab horses in southwest of Iran.

Microsporidiosisis anopportunistic infection frequently diagnosed in immunocompromised patients. Microsporidiosis causes symptomatic or asymptomatic infections immunocompromised/immunosuppressed patients may experience severe and disseminated microsporidiosis. Fecal samples were taken from 186 patients with human immunodeficiency virus (86) and cancer (100). Microsporidia spp. were detected 27.9% (24/86) and 11.6% (10/86) in HIV + patients, 18% (18/100) and 7% (7/100) of cancer patients using polymerase chain reaction (PCR) and trichrome staining respectively. Enterocytozoon bieneusi was detected in 28(66.7%) patients, while Encephalitozoon intestinalis was detected in 10 (23.8%) patients. Encephalitozoon cuniculi was detected in only one patient, while Encephalitozoon hellem was detected in three patients. Microsporidia was significantly higher in patients with diarrhea than those without diarrhea (P = 0.001). CD4 T cells blood count was significantly lower (