Iranian Journal of Parasitology Tehran University of Medical Sciences en-US Iranian Journal of Parasitology 1735-7020 Immunodiagnosis of Visceral Leishmaniasis: Current Status and Challenges: A Review Article <p><strong><em>Background:</em></strong> Diagnosis of Visceral Leishmaniasis (VL) is still challenging. This review highlighted current status and challenges in the serological diagnosis of VL. Furthermore, the drawback of currently available serological tests and the most recent advancement in the designing and application of these assays for the diagnosis of VL are addressed.</p> <p><strong><em>Methods:</em></strong> All the published literature cited within PubMed, ISI Web of Science, Google Scholar, Scopus, and IranMedex, regarding the immunodiagnosis of VL in human were sought from 2000 till Mar 2017. The search terms were “visceral leishmaniasis”, or “kala-azar" subsequently combined with the search terms "diagnosis", "serodiagnosis", "human", "serological", "antigen detection" or "antibody detection". Data were extracted from literature which fulfilled our eligibility criteria.</p> <p><strong><em>Results:</em></strong> Direct agglutination test (DAT) and rk39 dipstick have made a great improvement in the serological diagnosis of VL. Besides, other kinesin-related protein including K26, K28, and KE16 provided promisingly diagnostic accuracy in the diagnosis of VL. The Latex Agglutination Test for the diagnosis of VL (KAtex), with moderate sensitivity but high specificity, made a substantial contribution to the field. Moreover, a range of protein antigens has recently been detected in the urine of VL patients with encouraging diagnostic value.</p> <p><strong><em>Conclusion: </em></strong>The suboptimal diagnostic accuracy of the currently available serological assays for the diagnosis of human VL necessitates further research and development in this field. Outcomes of immunodiagnostic tests based on recombinant antigens are favorable. These proteins might be the most appropriate antigens to be further evaluated and utilized for the diagnosis of human VL.</p> Bahador shahriari Rad Zahra Rezaei Mehdi MOHEBALI ##submission.copyrightStatement## 2018-09-23 2018-09-23 13 3 331 341 Epidemiology and Control of Leishmaniasis in the Former USSR: A Review Article <p><span style="font-family: Calibri;"><span style="font-size: medium;"><span style="text-decoration: underline;">BACKGROUND</span>. All types of the Old World’s leishmaniasis were endemic on the territoru of the ex-Soviet Republics in the Central Asia and Transcaucasia. Epidemiological situation was well under control during the USSR era, due to implementation of complex anti-leismaniasis measures. These interventions were dramatically stopped as a result of the collapse of the USSR in 1991. Within next years the incidence of leishmaniasis returned to the level of the 1950s threatening to reach epidemic proportions unless urgent measures are undertaking. </span></span></p> <p><span style="font-family: Calibri;"><span style="font-size: medium;"><span style="text-decoration: underline;">METHODS</span>. Most relevant publications on epidemiology and control of leishmaniases in the Republics of Central Asia&nbsp; and Transcaucasia&nbsp; of the ex-USSR were screened. Data from foreign publications, especially from countries with similar or close to that epidemiological situation in respect of leishmaniases were also included.</span></span></p> <p><span style="font-family: Calibri;"><span style="text-decoration: underline;"><span style="font-size: medium;">RESULTS</span></span><span style="font-size: medium;">. Epidemiological analysis of spatial distribution of leishmaniases in the ex-USSR revealed that the northern borderline of these diseases is determined by the distribution of the vectors (42-46</span><sup><span style="font-size: small;">0 </span></sup><span style="font-size: medium;">North latitude). Within the endemic area, the foci of different kinds of leishmaniasis are often overlapped thus calling for deployment of integrated measures. The anthroponotic cutaneous leishmaniasis (ACL) was reported in settlements and towns of Central Asia and Transcaucasia of the ex-USSR. The natural foci of cutaneous leishmaniasis were widespread in the desert of Turkmenistan, Uzbekistan, southern Kazakhstan, and southern Tajikistan. The northern boundary of the zoonotic cutaneous leishmaniasis (ZCL) area coincided with the northern boundary of the distribution of great gerbils – the main reservoir of this infection in the ex-USSR. Visceral leishmaniasis (VL) occurred in the Central Asian Republics and in the republics of the Transcaucasia. The strategies of control and prevention of leishmaniases in the ex-USSR were based on good knowledge of epidemiology of infections. Holistic approach was adopted by the programs targeting the source of infection, vector(s) and man. </span></span></p> <p><span style="font-family: Calibri;"><span style="font-size: medium;"><span style="text-decoration: underline;">CONCLUSION</span>. The presence rise in the number of cases of different types of leishmaniasis in the ex-USSR strongly necessitates that health authorities should consider these diseases as an important public health problem. The immediate task would be re-building a comprehensive surveillance system consisting of active and passive case detection mechanism along with immediate treatment of the patients. </span></span></p> Vladimir SERGIEV Anatoly KONDRASHIN Sergei LITVINOV Lola MOROZOVA Natalia TURBABINA Ekaterina STEPANOVA Maria MAKSIMOVA Sergei SHEVCHENKO Evgeny MOROZOV ##submission.copyrightStatement## 2018-09-23 2018-09-23 13 3 342 350 Molecular Identification and Phylogenetic Classification of Leishmania spp. Isolated from Human Cutaneous Leishmani-asis in Iran: A Cross-sectional Study <p><strong><em>Background:</em></strong> In Iran, both forms of cutaneous (CL) and visceral leishmaniasis (VL) have been reported; so the accurate species identification of the parasite(s) and the analysis of genetic diversity are necessary.</p> <p><strong><em>Methods:</em></strong> The smears were collected from lesions samples of 654 patients with CL, who attended local health centers in 12 provinces of Iran during 2013-2015. The smears were checked for the presence of amastigotes by light microscopy. DNA of 648 <em>Leishmania</em> isolates, amplified by targeting a partial sequence of ITS (18S rRNA–ITS1–5.8S rRNA–ITS2) gene. Twenty-five of all the amplicons were sequenced and analyzed with restriction fragment length polymorphism (RFLP) using the <em>Taq</em>1 enzyme.</p> <p><strong><em>Results:</em></strong> All the smears were positive microscopically. The PCR-RFLP analysis revealed that 176 (27%) CL patients were infected with <em>L. tropica </em>and, 478 (73%) with <em>L. major</em>. The dominant species in all over Iran is <em>L. major</em>. The sequencing results of all CL patients and RFLP analysis confirmed each other. Based on our phylogenetic tree, 25 ITS DNA sequences were grouped into two clusters representing <em>L. major </em>and<em> L. tropica</em> species. Phylogenetic tree derived from the ITS sequences supports a clear divergence between <em>L. major</em> from the other species.</p> <p><strong><em>Conclusion:</em></strong> Discrimination of Iranian Leishmania isolates using ITS gene gives us this opportunity to detect, identify, and construct the phylogenetic relationship of Iranian isolates.</p> Anita MOHAMMADIHA Abdolhossein DALIMI Mehdi MOHEBALI Iraj SHARIFI Mohammadreza MAHMOUDI Asad MIRZAEI Adel SPOTIN Mahmoodreza BEHRAVAN Mehdi KARIMI Mohsen ARBABI Shahram NEKOEIAN Reza KALANTARI Behzad GHORBANZADEH ##submission.copyrightStatement## 2018-09-23 2018-09-23 13 3 351 361 New Nodule Type Found in the Lungs of Pomacea canaliculata, an Intermediate Host of Angiostrongylus cantonensis <p><strong><em>Background:</em></strong> <em>Pomacea canaliculata</em> (<em>P</em><em>.</em><em>canaliculata</em>) lung nodules, were commonly caused by <em>Angiostrongylus cantonensis </em>infection. Here, we found a new nodule type without any parasites.</p> <p><strong><em>Methods: </em></strong>Overall, 447 P. canaliculata snails were collected in Ning Bo, Zhe Jiang, China in 2018. In order to exhibit the similarities and differences between two nodules types (2018, Huzhou Zhejiang, China), both types were collected in formalin for tissue pathological sectioning. Besides, to obtain the microbial community of the new nodule, the 18S ribosomal RNA (rRNA) gene of it was amplified and analyzed using the Illumina second-generation sequencing platform.</p> <p><strong><em>Results: </em></strong>Although two nodules were found in the lungs of <em>P</em><em>. </em><em>canaliculata</em>, they were different in shape and pathology. Illumina sequencing indicated <em>Poterioochromonas </em>sp., a species of golden algae, might be the causing agent of the new nodule.</p> <p><strong><em>Conclusion: </em></strong>We firstly found a new pathological nodule type in the lungs of <em>P</em><em>. </em><em>canaliculata</em>, and this nodule might be induced by golden algae infection, however, the direct link between the golden algae and the new nodules, as well as the nodules’ impact on the snails’ physiology and <em>A</em><em>. </em><em>cantonensis</em> infection require further study.</p> Yue GUO Hong Chang ZHOU Ying DONG Ting ZHANG Yu Yang SUN Jian Feng ZHONG Yu Liang CAO Sheng Wen SHAO Yong Liang PAN Hai Yan DONG ##submission.copyrightStatement## 2018-09-23 2018-09-23 13 3 362 368 An Experimental Model of Primary Amoebic Meningoence phalitis Due to Naegleria australiensis in Iran <p><strong><em>Background:</em></strong> The main aim of the present research was to develop the experimental meningo encephalitis due to <em>Naegleria australiensis</em> isolated from geothermal water sources in mice model, November 2017 in Iran.</p> <p><strong><em>Methods:</em></strong> <em>Naegleria australiensis</em> was isolated from geothermal water sources in northern Iran. The number of amoebae was adjusted to be 1×10<sup>4</sup>/ml amoebae. The experimental infection was done using 3 wk old male (BALB/c) mice. Seven animals were used for experimental amebic infection and one animal was selected for the control. Intranasal (IN) and intracerebral (IC) inoculation of amoebae were done. The mice were then monitored on daily observation and as soon as they present any brain involvement they sacrificed. The brain of all animals was then dislocated and passaged in non-nutrient agar.</p> <p><strong><em>Results:</em></strong> One mouse out of seven infected mice were showed clinical symptoms of meningoencephalitis. Within few hours of culture of the brain, many vegetative forms of amoebae were detected in plate culture. The other infected animals and control mice showed no clinical symptoms until day 14. After 14 d all the animals sacrificed. The culture was negative up to one month.</p> <p><strong><em>Conclusion:</em></strong> The lack of brain involvement of other animals in the present study could be due to animal immune system or it may be possible that the amoebae did not reach to olfactory bulb of nostrils.</p> Alireza LATIFI Maryam NIYYATI Seyyed Javad SEYYED TABAEI Farid TAHVILDAR BIDEROUNI Ali HAGHIGHI Zohreh LASJERDI ##submission.copyrightStatement## 2018-09-23 2018-09-23 13 3 369 372 Detection of Toxoplasma gondii in Acute and Chronic Phases of Infection in Immunocompromised Patients and Pregnant Women with Real-time PCR Assay Using TaqMan Fluorescent Probe <p><strong><em>Background:</em></strong> <em>Toxoplasma gondii</em>, cause severe medical complications in infants and immune-compromised individuals. As using early, sensitive and rapid technique has major in diagnosis of toxoplasmosis, the present study was aimed to detect parasite by using from repetitive element (RE) and B1genes, in blood samples of seropositive immunocompromised patients and pregnant women.</p> <p><strong><em>Methods:</em></strong> A total of 110 peripheral blood samples were collected from seropositive cases with anti-<em>T. gondii</em> antibodies, including immunocompromised patients and pregnant women. DNA was extracted by a commercial kit and subjected to TaqMan probe-based real-time PCR assay by using primers and probes specific for RE and B1 genes, separately. The data were analyzed by Kappa test and SPSS-22 software.</p> <p><strong><em>Results:</em></strong> In the pregnant women, 17 (68%) and 14 (56%) samples from 25 IgM+/ IgG+ cases and, 7 (25%) and 6 (21.4%) samples from 28 IgG+/IgM- cases were positive by RE and B1 real time PCR, respectively. Likewise, in immunocompromised group, 20 (66.6%) and 17 (56.6%) samples from 30 IgM+/ IgG+ cases and 2 (7.4%) and 2 (7.4%) samples from 27 IgG+/ IgM- cases were positive by RE and B1 real time PCR, respectively.</p> <p><strong><em>Conclusion:</em></strong> Probe-based real time PCR assay is a quantitative approach for early diagnosis of <em>T. gondii</em> infection in clinical samples. Moreover, this method can be more appropriate in diagnosis of acute and reactivated toxoplasmosis. In addition our results indicated that RE gene is more sensitive than B1 gene.</p> Parisa MOUSAVI Hossein MIRHENDI Mehdi MOHEBALI Saeedeh SHOJAEE Hossein KESHAVARZ VALIAN Shirzad FALLAHI Setareh MAMISHI ##submission.copyrightStatement## 2018-09-23 2018-09-23 13 3 373 381 The First survey of isolation and molecular typing of Toxoplasma gondii by bioassay and PCR method in BALB/C mice in camels from eastern Iran <p><strong><em>Background</em></strong><strong>:</strong>This study was designed to isolation of Toxoplasma gondii from camels by bioassay method in mice model and detect parasitic DNA in brain mice by molecular methods.</p> <p><strong><em>Methods</em></strong><strong>:</strong>A total of 100 tissue samples including heart(n=50),and diaphragm(n=50) were obtained from camels(n=50) slaughtered in abattoirs from three provinces located in eastern Iran.Bioassay method was done in positive MAT camels and Nested PCR performed in tissue samples to amplify the B1 and GRA6 genes. The existence of polymorphic restriction sites for endonuclease MseI was used with PCR-RFLP method and Sequencing analysis to evaluate the prevalence of type strains (I, II and III)</p> <p><strong><em>Results:</em></strong> Totally,13(26 %) tissue samples of camels were found positive for the T. gondii B1 gene, including 7(14%) diaphragm,6(12%) heart. Moreover,3(6%) tissue samples of camels were found positive with GRA6 gene for T. gondii. There are three genotypes and mix genotype using MseI enzyme among all positive samples.</p> <p><strong><em>Conclusion:</em></strong> The obtained results for the first time demonstrated the presence of T. gondii DNA in the tissues of camels from east of Iran. Since consumption of meat camels are raising in Iran, there may be a high risk of contamination through consumption of products from these hosts due to their susceptibility to the infection.</p> <p><strong>&nbsp;</strong></p> Amir TAVAKOLI KARESHK Razieh TAVAKOLI OLIAEE Hossein MAHMOUDVAND Amir KEYHANI Hossein MAHMOUDVAND Amir KEYHANI Mohammad Ali MOHAMMADI Hossein MAHMOUDVAND Mehdi BAMOROVAT Mohammad Ali HAJHOSSEINI Naser ZIA-ALI ##submission.copyrightStatement## 2018-09-23 2018-09-23 13 3 382 391 Expression of Plasmid Encoded GRA4 Gene of Toxoplasma gondii RH Strain in CHO Eukaryotic Cells <p><strong><em>Background:</em></strong> Toxoplasmosis is a common infection all around the world. During pregnancy; it may lead to congenital disorders or abortion in human and animals. Severe damage of toxoplasmosis indicates to require effective vaccine. One of dense granules antigen is GRA4 that secrete from tachyzoite and bradyzoite. GRA4 genome is unique without intron and is one of the major immunogenic proteins from <em>Toxoplasma gondii. </em></p> <p><strong><em>Methods:</em></strong> We confirmed the cloning of GRA4 gene into pcDNA3 by restriction enzyme and PCR of GRA4 gene with pcGRA4 plasmids as template. Then with using calcium- phosphate method we transfected the pcGRA4 into CHO (Chinesehamster ovary) cells. The yielded protein was separated by SDS-PAGE and moved by electroblotting to nitrocellulose paper.</p> <p><strong><em>Results:</em></strong> Result of SDS-PAGE analysis showed the appearance of band approximately 42 kDa which was absent in the negative control, that was able to identify toxoplasmosis antibody IgM<sup>+</sup> serum in western blot analysis.</p> <p><strong><em>Conclusion: </em></strong>pcGRA4 plasmid is able to synthesis of antigenic protein in CHO cells. The ability of pcGRA4 for induction of protective immune response against toxoplasmosis will be evaluated in mouse model.</p> Marjaneh AGHDASI Fatemeh GHAFFARIFAR Fatemeh FOROOGHI Abdol Hossein DALIMI ASL Zohre SHARIFI Nahid MASPI ##submission.copyrightStatement## 2018-09-26 2018-09-26 13 3 392 398 Comparative Efficacy of Diethylcarbamazine, Nitazoxanide and Nanocomposite of Nitazoxanide and Silver Nanoparticles on the Dehydrogenases of TCA Cycle in Setaria cervi, in Vitro <p><strong><em>Background:</em></strong> Bovine filariid, <em>Setaria cervi </em>may cause serious pathological condition such as cerebrospinal nematodiasis in sheep, goat and horses. Since TCA cycle enzymes have certain biological functions that make them essential for the survival of parasite and therefore, efficacy of diethylcarbamazine (DEC), nitazoxanide (NTZ) and a nanocomposite of nitazoxanide and silver nanoparticles (NTZ+AgNPs) was assessed on succinate, malate and isocitrate dehydrogenases in the microfilariae (mf) and adult <em>S. cervi</em> worms.</p> <p><strong><em>Methods: </em></strong>This study was conducted in the Department of Zoology, Aligarh Muslim University, Aligarh, India during 2015-2016. Adult and microfilariae of <em>S. cervi</em> were incubated in 100 mg/ml of DEC, NTZ, and NTZ+AgNPs for 24 and 6 h, respectively at 37 °C. Succinate, malate and isocitrate dehydrogenases were localized by putting the mf and adult worms in the incubating medium containing their respective substrates at 37 °C for 2 h followed by counterstaining in 2% methylene green for 15 min.</p> <p><strong><em>Results:</em></strong> Maximum inhibition of TCA cycle enzymes was observed in both microfilariae and adult worms treated with nanocomposite of NTZ-AgNPs. Ruptured sheath along with nanoparticles sticking to the body surface was noticed in NTZ+AgNPs treated microfilariae.</p> <p><strong><em>Conclusion: </em></strong>NTZ+AgNPs proved most effective synergistic combination against TCA cycle enzymes which blocked the isocitrate and malate dehydrogenase almost completely, and succinate dehydrogenase to large extent in both microfilariae as well as adult worms of <em>S. cervi</em>. AgNPs ruptured the sheath and allowed the NTZ to attach and penetrate the main body to exert maximum effect on the enzymes.</p> Sharba KAUSAR Wajihullah KHAN ##submission.copyrightStatement## 2018-09-23 2018-09-23 13 3 399 405 Induction of Apoptosis in Toxoplasma gondii Infected Hela Cells by Cisplatin and Sodium Azide and Isolation of Apoptotic Bodies and Potential Use for Vaccination against Toxoplasma gondii <p><strong><em>Background:</em></strong> <em>Toxoplasma gondii </em>can infect a wide range of mammalians, especially humans. It controls several intracellular signals for the inhibition of apoptosis. This study aimed to investigate the apoptogenic effect of cisplatin and sodium azide on <em>T. gondii </em>infected HeLa cells and isolate apoptotic bodies (blebs) as a potent stimulator of the immune system.</p> <p><strong><em>Methods:</em></strong><em> T</em>he cytotoxic properties of cisplatin and sodium azide (NaN3) on HeLa cells were evaluated by MTT assay. Moreover, the apoptogenic activity of cisplatin and NaN3 was studied using flow cytometry (Annexin V/PI double staining) and scanning electron microscopy (SEM). Finally, apoptotic bodies were separated by centrifugation<em>. </em></p> <h4><em>Results:</em> MTT assay data showed that the survival rate of cells treated with different concentration of NaN3 was significantly reduced, compared to negative control groups. Concerning cisplatin, only concentration of 20 μM had not a significant impact on the cell viability; however, the other concentration of cisplatin significantly reduced cell viability, compared to negative control groups. The level of early apoptosis in uninfected HeLa cells was higher compared to infected HeLa cells treated with cisplatin and NaN3. Finally, apoptotic bodies were separated from T. gondii infected HeLa cells treated with cisplatin.</h4> <h4><em>Conclusion:</em> Apoptosis was induced in both uninfected and infected HeLa cells with T. gondii and apoptotic bodies were isolated from infected cells. Therefore, further studies on apoptotic bodies are required in order to find a proper candidate for vaccine preparation against <em>T. gondii</em> infections.</h4> Kourosh CHERAGHIPOUR Laleh SHARIATI Hossein KHANAHMAD Mazdak GANJALIKHANI-HAKEMI Abbas MORIDNIA Mina MIRIAN Nader PESTEHCHIAN ##submission.copyrightStatement## 2018-09-23 2018-09-23 13 3 406 415 Parasitic Helminths in Wild Boars (Sus scrofa) in Mazandaran Province, Northern Iran <p><strong><em>Background:</em></strong> Wild boars (<em>Sus scrofa</em>) are distributed worldwide and found in many parts of Iran. Although <em>S. scrofa </em>is reservoirs for many parasites, there is little data on helminthic prevalence in them. We aimed to survey the status of helminthic infections in <em>S. scrofa</em> in the Mazandaran Province of northern Iran.</p> <p><strong><em>Methods:</em></strong> Twenty-one wild boars were captured and examined for helminth infection during Dec 2012-Mar 2014. Adult worms such as <em>Macracanthorhynchus hirudinaceus</em> were identified by helminth size and shape, and the arrangement of the proboscis hooks. The sedimentation and flotation techniques were used to detect parasite eggs and larvae in faecal samples. Muscle samples were also surveyed for <em>Trichinella</em> larvae by artificial digestion method.</p> <p><strong><em>Results:</em></strong> Of the 21 samples, 13 (61.9%) were infected with one or more helminth species. Seven helminth types were identified in the alimentary track, comprising 5 nematodes, 1 trematode, and 1 acanthocephalan, with prevalence rates of <em>Macracanthorhynchus hirudinaceus</em> (57.14%)<em>, Globocephalus </em>spp<em>.</em> (33.33%)<em>, Trichuris suis</em> (19.04),<em> Gongylonema pulchrum</em> (14.28%), <em>Fasciola hepatica</em> (14.28%), <em>Dioctophyma</em><em> renale</em> (4.76%), and <em>Ascaris suum</em> (4.76%).</p> <p><strong><em>Conclusion:</em></strong> Wild boars might be involved in transmitting zoonotic parasites to humans. The abundance of these animals near human habitation creates favorable conditions for infection. So the risk of parasitic helminth diseases increases in other animals and humans.</p> Samira DODANGEH Davoud AZAMI Ahmad DARYANI Shirzad GHOLAMI Mehdi Sharif Iraj MOBEDI Shahabeddin SARVI Eissa SOLEYMANI Mohammad Taghi RAHIMI Majid PIRESTANI Shaban GOHARDEHI Reza BASTANI ##submission.copyrightStatement## 2018-09-23 2018-09-23 13 3 416 422 Genetic Identification of Echinococcus granulosus Isolates in Hamadan, Western Iran <p><strong><em>Background:</em></strong> Cystic echinococcosis is a zoonotic infection and considered as a major economic and public health concern worldwide. This research was conducted to determine genotypic characteristics of livestock and human hydatid cyst isolates from Hamadan area, western Iran.</p> <p><strong><em>Methods:</em></strong> Sampling was conducted in Hamadan industrial slaughterhouse and Beast Hospital of Hamadan City, western Iran, from 2015 to 2016. Overall, 74 livestock isolates including 69 sheep, 3 cattle and 2 goats and 9 human hydatid cysts were genotyped by PCR amplification of the rDNA ITS1 region and followed restriction fragment length polymorphism (RFLP) analysis with four restriction endonuclease enzymes, <em>RsaI</em>, <em>HpaII</em>, <em>AluI</em>, and<em> TaqΙ</em>, and sequencing.</p> <p><strong><em>Results:</em></strong> The PCR amplicon size of each isolate was approximately 1 kb which was the same with that of sheep strain. According to the RFLP patterns, the isolates belonged to a single species, <em>E. granulosus</em> sensu stricto (G1–G3 complex). Furthermore, sequencing of representative amplicons confirmed that the RFLP-genotyped isolates corresponded to <em>E. granulosus</em> sensu stricto.</p> <p><strong><em>Conclusion:</em></strong> <em>E. granulosus</em> sensu stricto is the prevailing species of <em>E. granulosus </em>sensu lato in the region and pointed out the importance of sheep/dog cycle in <em>human transmission</em><em>. </em></p> Mohammad MATINI Maliheh ROOSTAEI Mohammad FALLAH Amir Hossein MAGHSOOD Massoud SAIDIJAM Majid FASIHI HARANDI ##submission.copyrightStatement## 2018-09-23 2018-09-23 13 3 423 429 Molecular and Serological Detection of Toxoplasma gondii in Stray Cats in Shiraz, South-central, Iran <p><strong><em>Background:</em></strong> Toxoplasmosis is a global zoonotic disease that causes critical medical complications in neonates and immunocompromised persons. Infection rates in cats, specifically stray cats, are believed to be the best sentry of the level of <em>Toxoplasma gondii</em> in the environment. Therefore, in this study, we surveyed <em>T. gondii</em> infection in stray cats of Shiraz, one of the metropolises of Iran.</p> <p><strong><em>Methods:</em></strong> The appearance of antibodies and DNA of <em>T. gondii</em> in samples from 145 stray cats was determined in order to appraise the prevalence of <em>T. gondii</em> infection, by MAT and Nested-PCR.</p> <p><strong><em>Results:</em></strong> The rate of <em>T. gondii</em> infection in the cats was 69% by PCR and 82.8% by MAT. Besides, the highest rate of infection was discerned in diaphragm (37.9%) and intercostal muscle (34.5%), while the lowest rate was related to ileum (6.9%). Moreover, the similarity between MAT with titers 1:20, 1:40 and PCR were 79.2% and 86.2%, respectively (<em>P</em>=0.02 and <em>P</em>=0.0001).</p> <p><strong><em>Conclusion</em></strong><strong><em>:</em></strong> Nested-PCR and MAT are valuable techniques for molecular and serological detection of <em>T. gondii</em>. The prevalence of <em>T. gondii</em> infection in stray cats in Shiraz is high.</p> Qasem ASGARI Iraj MOHAMMADPOUR Razieh PIRZAD Mohsen KALANTARI Mohammad Hossein MOTAZEDIAN Shahrbanou NADERI ##submission.copyrightStatement## 2018-09-23 2018-09-23 13 3 430 439 Genetic Diversity in C-terminal of SERA5 Gene in the Blood Stage of Human Isolates of Plasmodium vivax in Sistan and Baluchistan, Iran <p><strong><em>Background:</em></strong> <em>Vivax</em> malaria is more prevalent in the malarious areas of Iran, which makes vaccine research a high priority. Serine Repeat Antigens (SERA) have essential role in the parasite life cycle and high expression profiles of PvSERA5 make it suitable vaccine candidates. This study aimed to evaluate the genetic diversity of C-terminal region of PvSERA5 in Iranian isolates of <em>Plasmodium vivax</em> in Sistan and Baluchistan.</p> <p><strong><em>Methods: </em></strong>Totally, 49 blood samples were taken from symptomatic malaria patients in Sistan and Baluchistan Province in 2016. Mono-infection to <em>P. vivax</em> was confirmed by 18srRNA-Nested-PCR. Genomic DNA was extracted and C-terminal region of PvSERA5 was amplified by specific primers. PCR-products have been sequenced and analysis was done by using bioinformatics software, mainly DnaSP &amp; MEGA5.</p> <p><strong><em>Results: </em></strong>Genetic diversity was calculated 14.8% in C-terminal region of PvSERA5 in Iranian isolates, 19 different sequences and 4 haplotypes existed. The amount of Tajima’s D (0.3805) and ratio of non-synonymous to synonymous mutation (1.82) showed that C-terminal region of PvSERA5 is under positive natural selection; also intragenic recombination could interfere.</p> <p><strong><em>Conclusion:</em></strong> Results could be helpful in any research, regarding this antigen as vaccine candidate in Iran or worldwide.</p> Ahmad ABOLGHAZI Aliehsan HEIDARI Vahideh MOIN-VAZIRI Ali HAGHIGHI Seyyed Javad SEYYED TABAEI Hossein KESHAVARZ Saeedeh SHOJAEE ##submission.copyrightStatement## 2018-09-23 2018-09-23 13 3 440 447 Oxidant/Antioxidant Status, PON1 and ARES Activities, Trace Element Levels, and Histological Alterations in Sheep with Cystic Echinococcosis <p><strong><em>Background:</em></strong> Total antioxidant status (TAS), total oxidant status (TOS) and oxidative stress index (OSI), nitric oxide (NO), zinc (Zn), copper (Cu) levels, paraoxonase (PON1), arylesterase (ARES) activities, and biochemical changes were studied on sheep with cystic echinococcosis.</p> <p><strong><em>Methods:</em></strong> The materials were taken from 2-3 yr old sheep slaughtered in Van Province, Turkey in 2017. Before the slaughter, blood samples were collected from the healthy sheep, while various organs of animals were examined for hydatid cysts after the slaughter. Thirty sheep were protoscolex positive, hydatic group, while 30 sheep that did not have any pathological lesions in organ examinations were accepted as the control group. TOS levels, PON1 and ARES activities, and Zn levels were determined by commercial kits, while Cu levels were determined by atomic absorption spectrophotometer. The collected data were then statistically analyzed.</p> <p><strong><em>Results:</em></strong> Serum TOS and OSI levels were significantly higher in sheep with cystic echinococcosis compared to the control group (<em>P</em>&lt;0.001). TAS levels (<em>P</em>&lt;0.01), PON1 and ARES activities, on the other hand, were significantly higher in control group compared to the cystic echinococcosis group (<em>P</em>&lt;0.001). There were no significant differences in Zn, NO and Cu levels between the groups.</p> <p><strong><em>Conclusion:</em></strong> PON1 and ARES activities increased in sheep infected with cyst hydatid. The decline of antioxidant reserves in the metabolism results in excessive amounts of free radicals, along with alterations of the normal histological structure of the cystic organ and changes in trace element metabolism.</p> Kıvanç IRAK Burçak Aslan ÇELİK Zelal KARAKOÇ Özgür Yaşar ÇELİK Nihat MERT Nihat Mert Mustafa Oğuzhan KAYA ##submission.copyrightStatement## 2018-09-23 2018-09-23 13 3 448 456 Genotypes of Enterocytozoon bieneusi in Dogs and Cats in Eastern China <p><strong><em>Background:</em></strong> <em>Enterocytozoon bieneusi </em>is a common opportunistic pathogen found in both humans and animals. As companion animals live in close contact with human being, they may act as a zoonotic reservoir and play an important role in transmitting this parasite to humans. We evaluated the prevalence, genotypic diversity and zoonotic potential of <em>E. bieneusi</em> in dogs and cats in <a href="">eastern China</a> during Apr to Dec 2013.</p> <p><strong><em>Methods: </em></strong>Fecal specimens from 315 dogs and 143 cats from veterinary hospitals in eastern China were examined in 2015 by internal transcribed spacer (ITS)-based PCR.</p> <p><strong><em>Results: </em></strong><em>E. bieneusi</em> was detected in 8.6% of canine and in 1.4% of feline samples. Seven genotypes of <em>E. bieneusi</em> were identified, including four known genotypes (PtEb IX, EbpC, Type IV and D) and three novel genotypes, named CHD1, CHD2 and CHD3. The dominant genotype in dogs was PtEbIX (59.3%; n=16/27). Five (CHD1, EbpC, CHD2, D and Type IV) of the seven genotypes were in the so-called zoonotic group 1, whereas genotypes PtEbIX belonged to the dog-specific group and genotypes CHD3 were placed in group 2.</p> <p align="left"><strong><em>Conclusion:</em></strong> Dogs are predominately infected with host-specific genotypes of <em>E. bieneusi</em>, and the finding of several zoonotic genotypes in dogs and cats reminds us of potentially zoonotic transmission of microsporidiosis.</p> Wen-Chao LI Jie QIN Kai WANG You-Fang GU ##submission.copyrightStatement## 2018-09-24 2018-09-24 13 3 457 465 Exploration of Sarcoptes scabiei Antigenic Protein Which Play Roles in Scabies Pathogenesis in Goats and Rabbits <p><strong><em>Background:</em></strong> Scabies or mange is an infectious skin disease caused by the mite <em>Sarcoptes scabiei</em>. This skin disease affects various livestock such as goats, sheep, swine, cattle, other animals like dogs, cats, wild animals and also affect human. This research aimed to explore the protein in mites <em>S. scabiei</em> which has antigenic character and play roles in scabies pathogenesis in goats and rabbits.</p> <p><strong><em>Methods:</em></strong> <em>S.scabiei</em> mites were isolated from goats and rabbits, and characterized using SDS-PAGE. In addition the protein was also analysed using Western Blot assay. The isolation and identification were carried out in 2015 at the Parasitology Laboratory of Veterinary Medicine Faculty, Universitas Airlangga, Surabaya, Indonesia.</p> <p><strong><em>Results:</em></strong> The identification results using SDS-PAGE of mites <em>S. scabiei</em> var. <em>caprae</em> expressed 12 protein bands between 26,7 kDa and 205,8 kDa, continued by Western Blot showed 3 protein bands, after being reacted with blood serum from scabies infected goat, it could be identified antigenic protein with molecule weight 205.8 kDa, 57.3 kDa, and 43 kDa. While protein in mites <em>S. scabiei</em> var.<em> cuniculi</em> identified 9 protein bands between 24 kDa and 75 kDa by SDS-PAGE, and the Western Blot assay identified antigenic protein with molecule weight 62 kDa and 51 kDa.</p> <p><strong><em>Conclusion:</em></strong> The antigenic protein of <em>S. scabiei</em> var. <em>caprae</em> and<em> S. scabiei</em> var. <em>cuniculi</em> showed that they are probably involved in the scabies pathogenesis in goats and rabbits.</p> Nunuk Dyah Retno LASTUTI Poedji HASTUTIEK Lucia Tri SUWANTI Dony CHRISMANTO ##submission.copyrightStatement## 2018-09-24 2018-09-24 13 3 466 472 Morphometric Analysis of the Intestine in Experimental Coccidiosis in Broilers Treated with Anticoccidial Drugs <p><strong><em>Background:</em></strong> Coccidiosis causes morphologic alteration in intestinal mucosa resulting in reduction of absorptive surface. Anticoccidials used as feed additives may induce changes in the intestinal mucosa. This study was designed to assess intestinal morphometry in broilers infected with <em>Eimeria </em>under different anticoccidial treatments.</p> <p><strong><em>Methods:</em></strong> To evaluate the effect of salinomycin and amprolium+ethopabate on intestinal morphometry in broilers experimental coccidiosis, in Tehran, Iran in May 2015, fifty-four Ross 308 birds were randomly divided into two challenged and unchallenged groups at the age of 12 days. The birds were challenged with <em>Eimeria </em>field isolate at day 14. Different growth and parasitological parameters including weight gain, feed consumption, FCR, macroscopic lesion score and oocyst score were recorded 7 d post-inoculation. Histological sections from four main parts of intestine (anterior, middle, lower intestines and cecum) were prepared and analyzed. Villus width and length and total mucosal thickness were measured microscopically.</p> <p><strong><em>Results:</em></strong> Amprolium+ethopabate and salinomycin significantly reduced coccidiosis gross lesions in infected birds. Microscopically anticoccidial administration in the presence of infection has significantly increased the villus length while the presence of amprolium+ethopabate in the absence of infection has greatly increased the mucosal thickness and villi height in comparison to the control group.</p> <p><strong><em>Conclusion:</em></strong> Anticoccidials may induce some histological changes in the mucosa when there is no parasite to be affected. Some of these effects may be advantageous for the intestinal epithelium integrity and hence the birds’ performance.</p> Sedigheh NABIAN Fatemeh ARABKHAZAEL Parvaneh SEIFOURI Alireza FARAHANI ##submission.copyrightStatement## 2018-09-24 2018-09-24 13 3 493 499 Allelic Variations of Plasmodium vivax Apical Membrane Antigen-1 (Pv AMA-1) in Malarious Areas of Southeastern Iran Using PCR-RFLP Technique <p><strong><em>Background:</em></strong> Although <em>Plasmodium vivax</em> is usually known as benign malaria, some variations of the parasite can result in acute and sever infection. In this study we tried to determine some genetic variations in <em>Pv</em>AMA-1 antigen among the samples were collected form southeastern Iran.</p> <p><strong><em>Methods:</em></strong> About two ml blood samples were collected into EDTA pre-dosed tubes from 30 <em>P. vivax</em>–infected patients individually between 2011 and 2013. A Giemsa stained thick and thin blood film was prepared from each of the patients. A PCR-RFLP technique was employed using EcoR-1, Pvu-II and Hind3 restriction enzymes to determine the allelic variations of the antigen.</p> <p><strong><em>Results:</em></strong> A 1300bp gene corresponding to <em>Pv</em>AMA-1 was selected for the amplification process. Among the total cases identified in this study 90% showed similar bounds when exposed to the restriction enzymes. Nine isolates (accession numbers: KF435081-KF435083 and JF682785-JF682790) were identified and registered in Gene bank. Identity among isolates was more than 96% in nucleotide level. Dendrogram clarified a close relationship among the clusters in spite of geographical distribution of the parasite.</p> <p><strong><em>Conclusion:</em></strong> This study increased our data about prevalence and variation of <em>Pv</em>AMA-1 alleles amongst <em>P. vivax</em> isolates in southeastern parts of Iran where besides native population bears considerable Afghan and Pakistani immigrants.</p> Afsaneh MOTEVALLI HAGHI Sepide MORADI Mehdi NATEGHPOUR Gholamhossein EDRISSIAN ##submission.copyrightStatement## 2018-09-24 2018-09-24 13 3 473 479 Endoparasites of Jungle Cats (Felis chaus) and Their Patholog-ic Lesions <p><strong><em>Background:</em></strong> The jungle cats (<em>Felis chaus</em>) are native to Asia, and found in Iran. Although north of Iran has a wide distribution of jungle cats, there is not any data about prevalence of parasitic infections in the population of these cats.</p> <p><strong><em>Methods:</em></strong> From 2012 to 2015, seven specimens of the wild jungle cat (<em>Felis chaus</em>) from north of Iran, Mazandaran Province, northern Iran were collected and examined for their endoparasites and pathological lesions which were caused by the parasites.</p> <p><strong><em>Results:</em></strong> Parasitological evaluations showed several species of endoparasites in the small intestine of cats including nematode (<em>Toxocara cati</em>), trematode (<em>Alaria alata</em>), and cestode (<em>Mesocestoides lineatus</em>). All of the examined jungle cats were infected to parasitic infections. From the total number of 7 jungle cats <em>T. cati</em> was found in 6 cats, <em>A. alata</em> in 1 cat, and <em>M. lineatus</em> was recovered from 3 cats. Histopathological samples displayed necrosis, vacuolar degeneration, atrophy, destruction of tissue, hyperemia, and infiltration of inflammatory cells in the intestine tissue.</p> <p><strong><em>Conclusion:</em></strong> This study for the first reported <em>T. cati</em>, <em>A. alata,</em> and <em>M. lineatus</em> and their pathologic effects in the small intestine of jungle cat.</p> Seyed Reza TABARIPOUR Mohammad Reza YOUSSEFI Seyed Mohammad HOSSEINI ##submission.copyrightStatement## 2018-09-24 2018-09-24 13 3 480 485 Effect of Origanum vulgare Hydroalcoholic Extract on Giardia lamblia Cysts Compared with Metronidazole in Vitro <p><strong><em>Background:</em></strong> Giardiasis, an intestinal infection, is made by the flagellate protozoan and on the other hand, positive effects of plants derivatives, especially phenolic derivatives, against giardiasis. The effect of <em>Origanum vulgare</em> (OV) hydroalcoholic extract is still uninvestigated. Thus, this study was conducted to evaluate the effect of OV hydroalcoholic extract on <em>Giardia lamblia</em> cysts compared with metronidazole in vitro.</p> <p><strong><em>Methods</em></strong><strong>: </strong>The present experimental study was conduct­ed in 2015-2016 in the Laboratory of Department of Parasi­tology of Islamic Azad University (Abhar Branch, Abhar, Iran). Cysts separated from feces by Bingham procedure were calculated by using the Hemusytumetr method. Five hundred µl of concentrations of 10, 100 and 200 mg/ml of OV hydroalcoholic extract and also125 mg/kg of metronidazole were added to the purified cysts of giardia. Control group was treated with normal saline. Anti-Giardia activity was calculated by using the light microscope for 30, 60 and 120 min and after exposure to eosin stain.</p> <p><strong><em>Results:</em></strong> The results indicated anti-Giardia activity of OV hydroalcoholic extract and the best response was achieved at higher levels so that there were no significant differences among OV groups at levels of 200 mg/kg with metronidazole (<em>P</em>&gt;0.05).</p> <p><strong><em>Conclusion:</em></strong> The anti-Giardia activity of Origanum<em> vulgare </em>extract is may due to the presence of phenolic compounds present in it.</p> Jaber DAVOODI Saeid ABBASI-MALEKI ##submission.copyrightStatement## 2018-09-24 2018-09-24 13 3 486 492 Ovarian Cancer or Hydatidosis? A Case Report <p><em>Echinococcus granulosus </em>has been described as the common etiology of hydatid cysts in many parts of the world. A 54-yr-old female with lower abdominal pain referred to Gynecology Ward of Sari Imam Khomeini Hospital, Iran in 2016. Sonography was carried out and cysts in ovaries and liver were observed. The cysts of liver seemed to be hydatidosis but physicians were suspected about ovarian cystic mass. Anti-<em>Echinococcus </em>antibodies (ELISA) screen was positive. The operation was done on her and treatment by albendazole started one week before surgery and continued after discharge from the hospital. Pathology confirmed hydatidosis in ovary, also patient follow-up was performed for three months by abdominal CT scan that showed peritoan full of many small hydatid cysts. Uncommon locations for constitution of hydatid cysts such as ovary and peritoan often make the diagnosis very difficult. Hydatidosis is considered in differential diagnosis of any cysts of the entire body, especially in endemic countries such as Iran.</p> Anahita NOSRATI Eissa SOLEYMANI Lotfollah DAVOODI ##submission.copyrightStatement## 2018-09-26 2018-09-26 13 3 500 504 Can Giardia Infection Impair the Diagnostic Level of Fecal Calprotectin in Patients with Inflammatory Bowel Disease? A Case Report <p>Inflammatory bowel disease (IBD) is attributed to complex conditions of gastrointestinal tract that is frequently reported all over the world. Fecal calprotectin evaluation is described as a primary tool to screen IBD patients. There are reports showing the confounding role of some microbial agents in diagnostic levels of calprotectin. A 32-yr-old woman with symptoms like IBD/IBS (irritable bowel syndrome); admitted to IBD Clinic of Behbood Research Center for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences, Tehran, Iran, Jan 2017 for evaluation of the level of fecal calprotectin. In spite of high level of calprotectin, trophozoite of <em>Giardia intestinalis</em> was observed in direct examination of stool sample. Microbial pathogens can lead to false elevation of fecal calprotectin and misdiagnosis of gastrointestinal patients suspected to IBD.</p> Sara MOHAMMAD ALI GOL Hamed MIRJALALI Hamid ASADZADEH AGHDAEI Mohammad Reza ZALI ##submission.copyrightStatement## 2018-09-24 2018-09-24 13 3 505 509 Extracellular Vesicles: A More Layer of the Complexity of Hydatid Fluid Contents and Functions <p>No abstract</p> Yixia CHEN Juntao DING ##submission.copyrightStatement## 2018-09-24 2018-09-24 13 3 510 511 Comment on “Zoonotic and Non-zoonotic Parasites of Wild Rodents in Turkman Sahra, Northeastern Iran” <p>No abstract</p> Seyed Mahmoud SADJJADI ##submission.copyrightStatement## 2018-09-24 2018-09-24 13 3 512 514