Iranian Journal of Parasitology Tehran University of Medical Sciences en-US Iranian Journal of Parasitology 1735-7020 Host-parasite Responses Outcome Regulate the Expression of Antimicrobial Peptide Genes in the Skin of BALB/c and C57BL/6 Murine Strains Following Leishmania major MRHO/IR/75/ER Infection <p><strong><em>Background:</em></strong> Different background of immunity responses determine resistance or susceptibility of certain mouse strains to <em>Leishmania major </em>infection. Some have been well known previously. Antimicrobial peptides (AMPs) such as cathelicidins and defensins are unique fragments of innate immunity system with well-known effects against the invasion pathogens. Despite their outstanding roles and being of extensive cases of cutaneous leishmaniasis (CL) caused by <em>L. major</em>, they have been less studied in <em>Leishmania</em> fields. The aim of present study was to determine whether these components play a role in the protection of skin against <em>Leishmania </em>infections.</p> <p><strong><em>Methods:</em></strong> The animal model of <em>Leishmania</em> infection was established by the subcutaneous inoculation of 5×10<sup>6</sup>(parasites/ml) from the stationary phase of <em>L. major </em>promastigotes to BALB/c and C57BL/6 mice from January 2016 to August 2016 in Kerman Province, southeast of Iran. After 1, 3 and 7 d of post-infection (PI), the samples needed for detecting of the mRNA levels of mouse beta-defensin (mBD)-1, mBD2, mBD3, mBD4, mBD6, cathlin-related antimicrobial peptide (CRAMP), interleukin (IL)-10, IL-12 and parasite load were taken under standard methods.</p> <p><strong><em>Results:</em></strong> The findings related to cytokines profiles in BALB/c (­IL-10, ¯IL-12) and C57BL/6 mouse strains (¯IL-10, ­IL-12) demonstrated that immunity system has been accurately activated during CL caused by <em>L. Major </em>parasites. We also observed a significant up-regulation of all aforementioned AMPs genes in BALB/c mice at selected times compared to another strain.</p> <p><strong><em>Conclusion:</em> </strong>CL occurred in BALB/c mice in spite of the fact that the expression of AMPs was higher than the other strain. AMPs genes are well expressed to provide defense against the parasites that have increased and escaped from immunity system but cannot create an absolute protection.</p> Hamid DANESHVAR Amir TAVAKOLI KARESHK Iraj SHARIFI Alireza KEYHANI Arash ASADI Razieh TAVAKOLI OLIAEE ##submission.copyrightStatement## 2018-12-18 2018-12-18 13 4 515 523 Prevalence of Animal Fasciolosis and Specification of Fasciola spp. Isolated from Sheep, Goats and Cattle by Molecular Meth-od: Hamadan Province, West of Iran <p><strong><em>Background:</em></strong> Fascioliasis is a common disease among humans and animals. Having global distribution, disease is developed by hepatic trematodes, <em>Fasciola hepatica</em> and<em> F. gigantica</em>. The main objective of this research was determining the prevalence of <em>Fasciola </em>species in Hamadan livestock and identifying those using PCR-RFLP.</p> <p><strong><em>Methods:</em></strong> Overall, 13607 livestock livers in the slaughterhouse of Hamadan, west of Iran including 10846 sheep, 995 cattle and 1766 goats were examined in 2015. In addition, 75 <em>Fasciola</em> (41 worms from sheep, 22 worms from goats and 12 worms from cattle) were examined by PCR-RFLP method.</p> <p><strong><em>Results:</em></strong> Totally, 100 livers were infected to <em>Fasciola</em> species (total prevalence: 0.74%; sheep 0.5%, goats 1.4%, cattle 1.5%). In the molecular results, prevalence of <em>F. hepatica</em> was higher than (92.5% in the sheep) <em>F. gigantic</em>, and in the cattle 91.5% and in the goats was 54.5% <em>F. hepatica</em>. Genotyping identified species confirmed intermediated types as <em>F. hepatica</em>.</p> <p><strong><em>Conclusion:</em></strong> Prevalence of <em>Fasciola</em> in this province is not so high. Intermediate types identified by PCR-RFLP method determined as <em>F. hepatica</em> by nucleotide sequencing. Because of morphological differences and interspecies variety, the accurate identification of <em>Fasciola </em>species needs using nucleotide sequencing.</p> KobraKobra PIRI Massoud SAIDIJAM Amirhossein MAGHSOOD Mohammad MATINI Mohammad FALLAH ##submission.copyrightStatement## 2018-12-18 2018-12-18 13 4 524 531 A Novel Nano Magnetic Beads Dot ELISA Immunoassay and Its Application on the Detection of Giardia lamblia Coproanti-gen <p><strong><em>Background:</em></strong> The traditional basis of diagnosis is identification of <em>Giardia lamblia</em> trophozoites or cysts in the stool of infected patients. Recently the advent of more objective techniques like antigen detection methods has led to an increase in their use versus those that rely on subjective microscopic examination of fecal specimens for <em>Giardia</em> cysts may facilitate diagnosis of <em>G. lamblia</em> in stool specimens.</p> <p><strong><em>Methods:</em></strong> This cross-sectional study was carried out from Oct 2015 to Feb 2016 on patients admitted to Benha University Hospitals (Benha, Egypt) and outpatients of Theodor Bilharz Research Institute (TBRI) (Giza, Egypt). Purified <em>G. lamblia </em>cysts antigen was prepared by two-phase sucrose gradient technique. Polyclonal antibody against purified <em>G. lamblia </em>cysts antigen was prepared and labeled with horseradish peroxidase and Nano Magnetic Beads (NMB) to be used as detecting antibody. A total of 72 stool samples, 32 samples positive for giardiasis, 20 samples positive for other parasitic infections in addition to 20 negative samples were examined using dot ELISA and NMB dot-ELISA.</p> <p><strong><em>Results:</em></strong> The sensitivity of the traditional dot-ELISA was 81.3 % and it increased by using the NMB-dot-ELISA to be 96.9% in stool samples. Specificity of both techniques was 97.5%.</p> <p><strong><em>Conclusion:</em></strong> Diagnosis of <em>G. lamblia</em> by NMB-Dot-ELISA technique is sensitive, specific, rapid and easy to perform and interpret. In this study, using the nano-magnetic beads increased the sensitivity of the applied technique.</p> Nagwa SHABAN ALY Ibrahim BAYOUMI Rabab SELEM Manal KARDOUSH Gehan RASHED Ahlam MOHARAM ##submission.copyrightStatement## 2018-12-18 2018-12-18 13 4 532 540 Identification of Clinical Immunological Determinants in Asymptomatic VL and Post Kala-azar Dermal Leishmaniasis Patients <p><strong><em>Background:</em></strong> Visceral Leishmaniasis (VL) caused by protozoa belonging to the genus <em>Leishmania, </em>usually have anthroponotic mode of transmission and is issue of great public health importance in Indian subcontinent. Asymptomatic cases of VL and PKDL are subject of keen interest to find their role in the transmission of VL in epidemic areas. We evaluated the immunological cytokine determinants expressed in most clinical suspects of asymptomatic VL and PKDL (IL-10, IFN-γ, and TNF-α).</p> <p><strong><em>Methods:</em></strong> Eighty-four participants were included at RMRIMS, Patna, India in 2016-17 out of which 64 asymptomatic individual positive for rK-39, without sign and symptoms of VL; 15 PKDL patient’s with past history of VL and 5 endemic healthy subjects were recruited from VL endemic areas. DAT and quantitative assessment of plasma cytokines was determined from the blood samples collected in a plain and sodium-EDTA vacutainer respectively from the subjects.</p> <p><strong><em>Results:</em></strong> The mean level of IL-10 in DAT<sub>pos</sub><sup>LOW</sup> of asymptomatic VL and PKDL was significantly higher than endemic healthy (<em>P</em>&lt;0.05). The cytokine polarization index (IFN-γ versus IL-10) was significantly low in PKDL cases compared with asymptomatic VL cases in DAT<sub>pos</sub><sup>LOW</sup> titre (<em>P</em>&lt;0.05). This index was low again but statistically not significant in PKDL than in asymptomatic VL when TNF-α was considered against IL-10. The ratio of IFN-γ: IL-10 and TNF-α: IL-10 was observed decreased both in asymptomatic VL and PKDL cases than in healthy from endemic areas.</p> <p><strong><em>Conclusion:</em></strong> Collectively we surmise from our data that asymptomatic VL can also play an important role like PKDL in transmission of VL.</p> Ashish Kumar SINGH Vidya Nand Rabi DAS Ajay AMIT Manas R DIKHIT Vijaya MAHANTESH Akhilesh KUMAR Raj Kishore PANDEY Shyam NARYAN Bipin K SINGH Krishna PANDEY Neena VERMA Pradeep DAS Sanjiva BIMAL ##submission.copyrightStatement## 2018-12-18 2018-12-18 13 4 541 548 Experimental study on Plasmodium berghei-Anopheles stephensi-BALB/c mouse system; implications for malaria transmission blocking assays <p><em><strong>Background</strong>:Plasmodium berghei</em> is a rodent malaria parasiteand has been very valuable means in the progress of our understanding of the essential molecular and cellular biology of the malaria parasites. Availability of hosts such as mice and vectors such as <em>Anopheles stephensi</em> has made this parasite a suitable system to study the parasite-host and vector-parasite relationships. Numerous studies have described life cycle and parameters influencing maintenance of the parasite within the mice or the mosquito. In this paper we revealed more details and have addressed some parameters and points influence maintenance of various life stages of the parasite (merozoites, macrogametocytes, ookinetes, oocysts and sporozoites) in the laboratory model&nbsp;<em>P.berghei–A.stephensi</em>-BALB/c mouse. This study helps understanding the biology of vertebrate-parasite and mosquito-malaria interactions that may aid in the development of a new generation of drug/vaccine and vector-based measures for malaria control.</p> Hossein DEHGHAN Mohammad Ali OSHAGHI Seyed Hassan MOSA-KAZEMI Mohammad Reza ABAI Fatemeh RAFIE Mehdi NATEGHPOUR Habib MOHAMMADZADEH Leila FARIVAR Mulood MOHAMMADI BAVANI ##submission.copyrightStatement## 2018-12-18 2018-12-18 13 4 549 559 Epidemiology of Hookworm Infection in the School-age Children: A Comparative Cross-sectional Study <p><strong><em>Background:</em></strong> Globally more than 740 million peoples are infected with hookworm. In sub-Saharan Africa, approximately 200 million people have been infected with hookworm, 90 million of them were children. The objective of this study was to identify the prevalence and determinant factors of hookworm infection in urban and rural school-age children’s.</p> <p><strong><em>Methods:</em></strong> This comparative cross-sectional study design was conducted in Bahir Dar and Mecha district, Bahir Dar, Ethiopia from Mar-May, 2014. Epi-info software was used to calculate the sample size. Multistage sampling technique was used to select the children’s. Blood and stool samples were collected from the children to determine the hemoglobin level and the presence of parasites. Data were entered into the computer using Epi-info software and transferred to SPSS for analysis. Descriptive statistics were used to identify the prevalence of hookworm and binary logistic regression was used to identify the determining factors for hookworm.</p> <p><strong><em>Results:</em></strong> The prevalence of hookworm was 22.3% [21%-24%]. Hookworm infection was associated with gender (AOR 1.31, 95% CI [1.03-1.66]), wearing shoe (AOR 0.35, 95% CI [0.25-0.48]), hand washing practice (AOR 0.62, 95% CI [0.48-0.79]), personal hygiene (AOR 0.45, 95% CI [0.34-0.61]), age (AOR 0.44, 95% CI [0.34-0.57]) or availability of latrine (AOR 0.08, 95 % CI [0.06-0.1]). Hookworm infection significantly decreases the school performance of children.</p> <p><strong><em>Conclusion:</em></strong> High prevalence of hookworm infection was observed. The ministry of health and ministry of education should include deworming activity as one strategy to increase quality of education.</p> Berhanu ELFU FELEKE ##submission.copyrightStatement## 2018-12-19 2018-12-19 13 4 560 566 Molecular Identification and Intra-species Variations among Leishmania infantum Isolated from Human and Canine Vis-ceral Leishmaniasis in Iran <p><strong><em>Background:</em></strong> In Iran, both forms of cutaneous (CL) and visceral leishmaniasis (VL) have been re-ported; 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 investigation was conducted from 2014 to 2015 in the northwest and south of Iran, where VL is endemic (7 provinces). Blood samples of patients and infected dogs were collected and sera separated for serologic examinations (DAT, rK39). Spleen or bone marrow samples from infected dogs were also collected to confirm the infection. DNAs of 70 samples amplified by targeting a partial sequence of ITS (18S rRNA–ITS1–5.8S rRNA–ITS2) gene. All the amplicons were sequenced and analyzed with restriction fragment length polymorphism (RFLP) using the TaqI enzyme.</p> <p><strong><em>Results:</em></strong> The cause of all 70 VL cases, were <em>L. infantum</em>, so, the dominant specie is <em>L. infantum</em>. The sequencing results of all VL cases and RFLP analysis corroborate each other. Discrimination of Iranian <em>Leishmania</em> isolates using ITS gene gives us this opportunity to detect, identify and construct the phylogenetic relationship of Iranian isolates. In addition, detection and differentiation of <em>Leishmania</em> spp. DNA was confirmed by amplification of variable area of the minicircle kDNA (conserved sequence blocks (CSB)).</p> <p><strong><em>Conclusion:</em> </strong>Low divergence and high likelihood were seen among <em>L. infantum</em> isolates of human and dogs from Iran with a very slight divergence was seen between isolates from northwest and south of Iran, thus grouped in a unique clad. No correlation was observed between intraspecies divergence and geographic distribution of the isolates.</p> AbdolhosseinAbdolhossein DALIMI Anita MOHAMMADIHA Mehdi MOHEBALI Asad MIRZAEI Mohammadreza MAHMOUDI ##submission.copyrightStatement## 2018-12-19 2018-12-19 13 4 567 576 Molecular Characterization of Echinococcus granulosus Sensu Lato from Livestock in North Khorasan Province, Iran <p><strong><em>Background:</em></strong> <em>Echinococcus granulosus</em> is one the most important zoonotic disease which is endemic in worldwide. Molecular method has allowed discrimination of different genotypes (G1-G10), providing new approach in development of prevention and control program of hydatid cyst. This study was conducted to identify the genotypes of <em>E. granulosus </em>from domestic animals in nine districts of North Khorasan Province using the mitochondrial cox1 gene sequence.</p> <p><strong><em>Methods: </em></strong>Overall, 122 hydatid cyst were collected during 2016-2017 from sheep (n=43) and cattle (n=79). DNA was extracted from protoscoleces and germinal layers and amplified by PCR. Phylogenetic analysis was also performed by analyzing the complete nucleotide sequences of mitochondrial cytochrome C oxidase subunit 1 (cox1) of <em>E. granulosus</em> genotypes from various locations.</p> <p><strong><em>Results: </em></strong>Sequencing of the amplified products revealed the presence of G1 as dominant genotype, G3 and <em>Echinococcus canadenesis </em>in one isolate each. Altogether, 9 haplotypes were detected based on cox1 gene. Haplotype 3 was the common variant that found in 58 including 42 cattle and 16 sheep.</p> <p><strong><em>Conclusion: </em></strong>This study provided knowledge on the identity of <em>E. granulosus </em>cysts collected from sheep and cattle in North Khorasan Province. Furthermore, these results showed the potentials of sheep as a main source of infection to humans, contributing the transmission and maintain of hydatid cyst in this region.</p> Mitra SALEHI Saeed YAGHFOORI Pejman BAHARI Mohsen SEYEDABADI Sima PARANDE SHIRVAN ##submission.copyrightStatement## 2018-12-19 2018-12-19 13 4 577 586 Effects of Cytokine Therapy for Treatment and Prophylaxis of Hydatidosis in Experimental Animal Model (Mice) <p><strong><em>Background:</em></strong> We aimed to use of IL-12 and IFN-γ for prevention and treatment of hydatidosis in experimental animal models.</p> <p><strong><em>Methods:</em></strong> This experimental study was conducted in Ahvaz City, southwest of Iran in 2017.&nbsp; Forty female BALB/C mice with 6-8 wk old were divided into four groups (two group test and two group control). The mice were challenged with 2000 protoscolices and a combination of IL-12 (10 µg/kg) and IFN-γ (25 µg/kg) was injected intraperitoneally daily one week at the time of challenging (prevention group) and three months after challenging (treatment group). After four months, the mice were euthanized and the number, size, and weight of the cysts were measured in all studied groups. Pathologic slides were prepared from cyst wall for pathologic changes study.</p> <p><strong><em>Results:</em></strong> There issignificant reduction in number, size, and weight of cysts in test groups.</p> <p><strong><em>Conclusion:</em></strong> Cytokine therapy is effective methods especially for prevention of cyst formation and to treat non-operable cases. Further study of combination of chemotherapy and cytokine therapy should be investigated.</p> Mahmoud RAHDAR Abdollah RAFIEI Rohollah VALIPOUR NOROUZI ##submission.copyrightStatement## 2018-12-19 2018-12-19 13 4 587 593 Comparison of Conventional and Molecular Diagnostic Techniques for Detection of Blastocystis sp. in Pig Faeces <p><strong><em>Background:</em></strong> <em>Blastocystis</em> is a common protist colonizing the gastrointestinal tract of humans and various animals. Pigs have been suggested to be a reservoir for human <em>Blastocystis </em>infections because of high prevalence of the parasite in these animals and the presence of zoonotic subtypes. Nevertheless, epidemiological data is often misinterpreted due to the lack of standard diagnostic procedures. This study aimed to compare the sensitivity of different diagnostic techniques in detection of <em>Blastocystis</em> sp. in pigs.</p> <p><strong><em>Methods:</em></strong> Overall, 48 individual faecal samples were collected from pigs reared in an intensive farming system (Autonomous Province of Vojvodina, Serbia) and were tested by microscopic examination of direct wet mount, in vitro cultivation in modified Jones' medium and conventional PCR for rRNA gene.</p> <p><strong><em>Results:</em></strong> Xenic in vitro cultivation in Jones’ medium showed higher sensitivity than direct wet mount when we compared it with PCR. Namely, the estimated sensitivity of direct wet mount was 46.15%, while the sensitivity of in vitro cultivation was 84.62%.</p> <p><strong><em>Conclusion:</em></strong> Low sensitivity of conventional parasitological compared to molecular methods is proven. Thus, reports on prevalence that rely solely on microscopy of faecal samples (unprocessed or concentrated) are probably underestimating the true prevalence of the parasite.</p> Tamás SÜLI Gordana KOZODEROVIĆ Aleksandar POTKONJAK Stanislav SIMIN Verica SIMIN Vesna LALOŠEVIĆ ##submission.copyrightStatement## 2018-12-26 2018-12-26 13 4 594 601 Survey and molecular characterization of trichomonads in pigs in Anhui Province, East China <p class="Default"><strong><em>Background: </em></strong>In pigs, several different trichomonad species such as <em>Tritrichomonas foetus</em>, <em>Tetratrichomonas buttreyi</em> and <em>Pentatrichomonas hominis</em> have been described as inhabiting the digestive tract. However, little information is available on the epidemiology of these neglected parasites in the Chinese pig population.</p> <p class="Default"><strong><em>Methods: </em></strong>The prevalence of <em>T. suis</em>, <em>T. buttreyi</em> and <em>P. hominis</em> among 500 fecal specimens from pigs at seven pigs farms inAnhuiProvince inChina was determined by PCR and DNA sequence analysis of the small subunit ribosomal RNA (SSU rRNA) genes.</p> <p class="Default"><strong><em>Results:</em></strong>The prevalence rates for <em>T. suis</em>, <em>T. buttreyi </em>and <em>P. hominis</em> were 2.8 % (14/500), 42.0 % (210/500) and 7.8 % (39/500), respectively. Mixed infections of two or three trichomonads were detected in 24 samples. The prevalence of the three trichomonads differed significantly between some age groups, with higher infection rates of <em>T. suis </em>and <em>T. buttreyi</em> in nursery pigs and <em>P. hominis </em>in preweaned pigs. The SSU rRNA sequences from <em>T. suis</em> and <em>P. hominis</em> showed 100 % homology with their respective homologous database sequences. However, we observed minor allelic variations in the SSU rRNA sequences from <em>T. buttreyi</em>, and the five representative sequence identified were named firstly as types 1, 2, 3, 4 and 5.Moreover, type 1 was found to be dominant in the present study.</p> <p class="Default"><strong><em>Conclusions:</em></strong>These findings highlight the potential risk posed by pigs in the transmission of trichomonad infections to humans and other animals.</p> Wen-Chao LI Kai WANG Yan LI Li-Ping ZHAO Yi XIAO You-Fang GU ##submission.copyrightStatement## 2018-12-19 2018-12-19 13 4 602 610 Frequency of Intestinal Parasites among Zoo Animal by Morphometric Criteria and First Report of the Bivitellobilhar-zia nairi from Elephant (Elephasmaximus maximus) in Iran <p><strong><em>Background:</em></strong> Intestinal parasitic infections are major causative agents of wildlife health complications among different parts of the world. This study aimed to investigate the gastro-intestinal parasites in feces of the zoo animals based on parasitological and morphometric criteria.</p> <p><strong><em>Methods:</em></strong> One hundred fresh fecal samples were collected from 35 species of animal lived in Eram park zoo, Tehran, Central Iran during Oct 2015 to Jun 2015. All collected samples were examined by microscopic observation following direct wet mount preparation (normal saline and Lugol's iodine), formalin-ether concentration, and permanent staining. The morphometric aspects of the recovered eggs were surveyed with the aid of Camera Lucida (×400).</p> <p><strong><em>Results:</em></strong> 65.7% (23/35) of zoo animal species were infected with intestinal parasites. The superfamily Trichostrongyloidea (6/16) and <em>Strongylus</em> sp. (16/4) were the most prevalent helminthic infections, while <em>Blastocystis</em> sp. (6/14), <em>Entamoeba</em> cyst (3/14) and <em>Eimeria</em> sp. (3/14) were the common protozoan parasites. For the first time, <em>Bivitellobilharzia nairi</em> egg was identified an elephant at Iran. Intestinal parasitic infections were apparently circulating among animals of the Eram park zoo.</p> <p><strong><em>Conclusion:</em> </strong>Identified parasitic infections can consider as a threatening source to visitors and workers' health that have contact with animals or their feces. Therefore, the effectual preventive strategies should be addressed to determine the risk factors, mechanisms of cross-transmission of parasite, the importance of applying the hygienic practices and well adjusting deworming programs for the animals, zoo workers and visitors.</p> Hamed KIANI Aliyar MIRZAPOUR Iraj MOBEDI Adel SPOTIN Seyyed Javad SEYYED TABAEI Mohsen RAHIMI ##submission.copyrightStatement## 2018-12-19 2018-12-19 13 4 611 617 Molecular Cloning, Expression and Characterization of Para Flagellar Rod Protein 1 of Trypanosoma evansi <p><strong><em>Background:</em></strong> Antigenic variation allows the trypanosomes to evade the potentially destructive host immune response and is an important reason for failure to develop a protective vaccine. Among the non-variant structural proteins, paraflagellar rod protein (PFR) is a prospective vaccine target owing to its role in the active movement of the parasite.</p> <p><strong><em>Methods:</em></strong> The PFR1 gene was cloned in pET-32a expression vector and after confirmation by restriction digestion, expressed as a Histidine-tagged fusion protein, in BL21 DE3 strain of <em>E. coli</em>. The expressed protein was affinity purified and then renatured. The immunoreactivity of the expressed recombinant protein was shown by western blot analysis using the specific serum. The experiment was carried out during 2013-14 at Division of Parasitology, Indian Veterinary Research Institute, Izatnagar, U.P., India.</p> <p><strong><em>Results:</em></strong> The results of sequencing, restriction digestion analysis, and PCR reaction revealed that cloning of PFR1 gene in pET-32a expression vector and the results of SDS PAGE and Western blot further confirmed its homogeneity and purity. The <em>in silico</em> Te-PFR1 (<em>T. evansi</em> PFR1) nucleotides sequence analysis revealed its close homology with the other members of the order Kinetoplastida.</p> <p><strong><em>Conclusion:</em></strong> We report here the molecular cloning, heterologous expression, and characterization of PFR1, a constituent protein of PFR. Due to its conserved nature, the PFR1 protein could be a prospective vaccine target against multiple <em>Trypanosoma</em> species.</p> Biswa RANJAN MAHARANA Anup KUMAR TEWARI Naduvanahalli RAJANNA SUDHAKAR Chinmoy MISHRA ##submission.copyrightStatement## 2018-12-19 2018-12-19 13 4 618 626 Investigation of the Prevalence of Fasciola hepatica in Small Ruminants in the Siirt Region, Turkey <p><strong><em>Background:</em></strong> Fasciolosis is a disease of the liver caused by trematodes in the family of Fasciolidae, particularly by <em>Fasciola hepatica</em> and <em>Fasciola gigantica</em>. The aim of this study was to investigate the prevalence of <em>F. hepatica</em> in sheep using the ELISA method, and in hair goats by post-mortem liver examination in the Siirt region, Turkey.</p> <p><strong><em>Methods:</em></strong> This study was conducted between Feb-Sep 2018. Five ml of blood samples were taken from the jugular veins of 320 sheep, which were selected from various locations of Siirt region by random sampling method. Fasciolosis seroprevalence in sheep was investigated by the ELISA method, using commercial kits (BIOK 211-Monoscreen AB ELISA <em>F. hepatica</em> test). In order to determine the prevalence of fasciolosis in hair goats, 580 slaughtered goats were examined for <em>F. hepatica </em>by incisions in the liver, gallbladder, and bile ducts.</p> <p><strong><em>Results:</em></strong> While 24 (7.50%) sheep were seropositive, 296 (92.50%) were seronegative. Regarding the hair goats, on the other hand, 82 (14.14%) were positive, while 498 (85.86%) were negative.</p> <p><strong><em>Conclusion:</em></strong> <em>F. hepatica</em> infection causes significant economic losses due to the destruction of the liver in small ruminants. Considering zoonotic properties of the disease, it has been concluded that the necessary measures should be taken and anti-helminthic drugs should be applied to the animals that come out of the pasture. Furthermore, periodic examinations should be conducted, and the breeders should be informed about the disease to raise awareness.</p> Özgür Yaşar ÇELİK Burçak ASLAN ÇELİK ##submission.copyrightStatement## 2018-12-19 2018-12-19 13 4 627 631 Frequency of Helminth Eggs in Faeces of Puppies Living in Urban or Rural Environments of Mexico City <p><strong><em>Background:</em></strong> The dog, <em>Canis familiaris</em>, a domestic animal that maintains close contact with humans and other animals, is considered as a potential source of zoonotic parasites. The current study aimed to determine the frequency of helminth eggs in feces of puppies of dog living in urban or rural environments of Mexico City between spring and summer of 2013.</p> <p><strong><em>Methods:</em></strong> Stool samples (n=180) were analyzed by sedimentation with formalin-ether. Samples were collected from puppies living in the urban zone (n=90; stray animals) or in the rural environment (n=90; stray animals, animals with owner and animals confined to a canine control center).</p> <p><strong><em>Results:</em></strong> Eggs of <em>Toxocara canis </em>(41%), <em>Ancylostoma</em> <em>caninum</em> (8%) and <em>Dipylidium caninum</em> (3%) were found in the rural environment but none in the urban zone. A frequency of 19% of <em>Toxocara</em> eggs was found in the canine control center, while, in stray puppies, the frequency was of 12% and 10% in animals with owner. Eggs of <em>Toxocara</em> were found in 33% samples of puppies with history of antiparasitic treatment.</p> <p><strong><em>Conclusion:</em></strong> This study supports the observation of helminth population reduction in urban environments. Further studies are needed to identify the factors that affect the development and transmission of helminth eggs in urban environments.</p> Jorge-Luis DE-LA-ROSA-ARANA Raquel TAPIA-ROMERO ##submission.copyrightStatement## 2018-12-22 2018-12-22 13 4 632 636 The Correlation between Serum Levels of Anti-Toxoplasma gondii Antibodies and the Risk of Diabetes <p><strong><em>Background:</em></strong> This study investigated the presence of specific antibodies against <em>Toxoplasma gondii</em> infection among people with diabetes (type I and II) in comparison with control group.</p> <p><strong><em>Methods:</em></strong> Overall 300 serum samples were collected equally from three groups including patients with type I and type II diabetes and non-diabetic healthy control that referred to Tabriz Central Laboratory in northwest Iran during July to Sep 2015. The level of specific IgG and IgM antibodies against <em>T. gondii</em> were measured using the chemiluminescence immunoassay (CLIA) method. Chi-square and One-Way ANCOVA were used for data analysis.</p> <p><strong><em>Results:</em></strong> Overall, 300 samples from diabetic patients (type I and type II) and control group were examined and results showed 3, 8 and 2 cases were seropositive for anti- <em>T. gondii</em> IgM respectively. Anti- <em>T. gondii</em> IgG seropositivity in type I and type II diabetes and control groups were 69%, 63% and 59% respectively. We did not observe any statistical differences among all studied groups in terms of toxoplasmosis. There was no statistically significant relationship between all variables and seropositivity for anti-<em>T. gondii</em> antibodies in type I and II diabetes and non-diabetic groups<strong>.</strong></p> <p><strong><em>Conclusion:</em></strong> Although there was no statistically significant relationship between diabetes and toxoplasmosis further investigations especially experimental studies using animal models are needed. Furthermore, these findings would not be contrary to the need for healthcare in order to the prevention of infectious disease in diabetic patients.</p> Maryam KHALILI Mahmoud MAHAMI-OSKOUEI Abbas SHAHBAZI Abdolrasoul SAFAIYAN Nader MOHAMMADZADEH-GHESHLAGHI Leyla MAHAMI-OSKOUEI ##submission.copyrightStatement## 2018-12-22 2018-12-22 13 4 637 642 In Vitro Assessment of Balanites aegyptiaca Fruit Methanolic Extract on the Adult Toxocara canis <p><strong><em>Background:</em></strong> The effectiveness of anthelmintics may diminish within approximately 10 yr of use relying on several factors such as anthelmintic resistance. This study aimed to assess in vitro effect of <em>Balanites aegyptiaca </em>fruits extract on the cuticle of adult worm of <em>Toxocara canis</em> as naturally alternative therapy.</p> <p><strong><em>Methods:</em></strong> <em>B. aegyptiaca</em> fruits were procured from the local markets in Aswan, Upper Egypt and authenticated at the Herbarium of National Research Centre. The effect of methanolic extract of <em>B. aegyptiaca</em> fruits on adult <em>T. canis </em>after 24 and 48 h incubating the parasites in Ringer solution containing 240 μg/ml <em>Balanites</em> extract was determined by scanning electron microscopy.</p> <p><strong><em>Results:</em></strong> The main changes induced by treatment with the tested extract were wrinkled cuticular surface and deformed sensory papillae. This cuticular distortion would undoubtedly disrupt its protective function and might be enough to expel <em>Toxocara</em> worms from dog’s intestine.</p> <p><strong><em>Conclusion:</em></strong> The use of this plant offers a chance for new nematocidal agent, which is economical alternative for the more expensive anthelmintics.</p> Hatem SHALABY Amira EL NAMAKY Omnia KANDIL Noha HASSAN ##submission.copyrightStatement## 2018-12-22 2018-12-22 13 4 643 647 The Taxonomic Position and Phylogenetic Relationship be-tween Digramma interrupta and Ligula intestinalis Based on Morphological and Molecular Diagnosis <p><strong><em>Background:</em></strong> The position of <em>Digramma interrupta</em> remains disputable as it was raised by Cholodkovsky from <em>Ligula alternans</em>. This study aimed to survey the evolutionary relationships and the taxonomic position of <em>D. interrupta</em> and <em>L. intestinalis</em>. It also intended to support or reject the validity of <em>D. interrupt </em>as an independent genus and its correlation with <em>L. intestinalis</em> on the basis of their morphological characteristics and a study on molecular data.</p> <p><strong><em>Methods:</em></strong> Overall, 1301 fish varieties, including 883 <em>Alburnoides bipunctatus</em> and 418 <em>Abramis</em> <em>brama</em>, were collected from north and north-western parts of Iran. <em>A. bipunctatus </em>samples were obtained from fresh water sources of the Maragheh dam (northwest) and the Ramesar Lake (north). Moreover, samples of <em>A. brama </em>were captured from the Aras Dam (northwest) and the Bandar-e-Anzali lagoon (north). PCR was used to generate a fragment spanning two independent ITS-inclusive parts: ITS1-5.8S and ITS2 with two pairs of primers.</p> <p><strong><em>Results:</em></strong> Nucleotide variation between <em>L. intestinalis</em> and <em>D. interrupta</em> samples amounts to about 3% to 7%. Between samples of <em>L. intestinalis</em> and GenBank data, and also between <em>D. interrupta</em> specimens and GenBank data, the diversity was seen for about 1% to 3%. Moreover, about 1% to 4% nucleotide variation was seen only in <em>L. intestinalis</em> samples caught from the same host, which could be supplementary to the presence of a species and/or strains in this genus.</p> <p><strong><em>Conclusion:</em></strong> Maybe <em>D. interrupta</em> was just a rare diplogonadic form of the <em>Ligula</em> species, not a different genus and not synonymous with the <em>Ligula</em> genus, but only another species of the <em>Ligula</em> genus.</p> Emad AHMADIARA Seyed Hossein HOSSEINI Fatemeh JALOUSIAN Hossein Ali EBRAHIMZADEH MOUSAVI ##submission.copyrightStatement## 2018-12-22 2018-12-22 13 4 648 654 Unusual Clinical Manifestations of Leishmania (L.) infantum chagasi in an HIV-coinfected Patient and the Relevance of ITS1-PCR-RFLP: A Case Report <p>Patients coinfected with Leishmania/HIV can develop atypical forms of visceral leishmaniasis (VL), making it indispensable to identify the etiological agent. We are presenting a postmortemspecie definition by ITS1-PCR-RFLP in a larynx tissue of a patient presented coinfectionLeishmania/HIV. This patient was from a leishmaniasis endemic region in São Paulo(SP), Brazil, and was diagnosed clinically with mucocutaneous leishmaniasis. Before a rK39immunochromatographic test positive, a tiny stored paraffin-embedded larynx tissue wasobtained post-mortem and submitted to 3 conventional PCR assays: kDNA (K20/K22 and<br>RV1/RV2), and ITS1 (LITSR/ L5.8S). The last one was followed by RFLP (HaeIII) andanalyzed by 4% Metaphor agarose gel electrophoresis. Leishmania genus and Leishmania(Leishmania) subgenus were defined by kDNA-PCR, with K20/K22 (120 bp) and RV1/RV2(145 bp), respectively. ITS1-PCR-RFLP identified L. (L.) infantum chagasi species visualizedby the restriction patterns of 180, 70 and 50 bp. This case draws attention to the necessityfor a clear identification of the etiological agent causing infection, especially in endemicregions of cutaneous and visceral leishmaniasis, and particularly in patients with comorbidities<br>who often present atypical forms of the disease. L. (L.) infantum chagasi, which is usuallyresponsible for VL, had changed its clinical spectrum for mucocutaneous. Unequivocalidentification was carried out by ITS-PCR-RFLP, therefore confirming rK39 result. Thesetechniques, which complemented each other, have a convenient cost-benefit ratio thatmakes them suitable to be applied in developing countries.</p> De Godoy NATALIA SOUZA DE Aiello VERA DEMARCHI de Souza REGINA MAIA Okay THELMA Braz LUCIA MARIA ALMEIDA ##submission.copyrightStatement## 2018-12-22 2018-12-22 13 4 655 660 Pediatric Vulvar Bleeding due to Leech Infestation: A Case Report <p>Leech infestation most frequently occurs in upper body cavities of children including pharynx, nose and esophagus and, more rarely, the vagina and vulva. Here we describe a 6-yr-old girl with vulvar bleeding caused by leech infestation that referred to the Emergency unit of the Booali Sina Hospital in Sari, Iran in Sep 2015. She had a history of swimming in a pond prior to the occurrence. The leech infestation particularly vulvar involvement among young girls is extremely rare and yet neglected event in the world.</p> Hajar ZIAEI HEZARJARIBI Mahdi FAKHAR Rezvan YALVEH Sayed Abdollah MOUSAVI Elham Sadat BANIMOSTAFAVI ##submission.copyrightStatement## 2018-12-22 2018-12-22 13 4 661 663 A 53-Year-Old Man with Intermittent Colicky Abdominal Pain due to Fasciola Incarceration in Common Bile Duct: A Case <p>Fascioliasis is a zoonotic disease caused by liver flukes of the genus <em>Fasciola</em>, as <em>F. hepatica,</em> and <em>F. gigantica</em>, mainly affecting the liver and biliary system during the chronic phase. These trematodes migrate through biliary ducts results in mild inflammation, when it is difficult to distinguish from obstructive lesions. Here we describe a 53-yr-old man from Golpayegan, a city in Isfahan Province, Iran, in year 2015, with occasional fever and chills, and also frequent colicky abdominal pain mainly on the right upper quadrant, with tenderness at that part. There was no jaundice and elevated bilirubin, but increased alkaline phosphatase was detected. Dilated common bile duct on abdominal sonography, without any visible lesion at its end and also dilated intra- and extrahepatic biliary ducts on abdominal CT-scan were seen. Endoscopic Retrograde Cholangiopancreatography (ERCP) detected incarceration of parasites behind Oddi's sphincter and also in common bile duct and serologic test (ELISA) confirmed fascioliasis. However, Iran is one of the most affected countries by <em>Fasciola</em>, being aware of rare symptoms and presentations of this disease can aid the physicians to make timely and accurate diagnosis and therefore reduce the consequent morbidities.</p> Davood YADEGARINIA Shabnam TEHRANI Zahra DOOSTI ##submission.copyrightStatement## 2018-12-22 2018-12-22 13 4 664 668 Wound Myiasis in a Flamingo (Phoenicopterus ruber) Caused by Calliphora spp. Larvae from Northwest of Iran: A Case Re-port <p>Myiasis is the infestation of animals or man tissues by parasitic dipterous fly larvae. Wound myiasis is the result of fly egg deposition on decaying flesh or pus discharging wounds. This case report describes a type of wound myiasis caused by <em>Calliphora</em> spp. in a Flamingo (<em>Phoenicopterus </em><em>ruber)</em> from East Azerbaijan Province, Iran. A 3-yr-old female Flamingo was suffering in its left wing leading to an extensive discharging wound, which was heavily infested by maggots (fly larvae). The examination of external morphological characters of the second and third-instar larvae, posterior spiracles and internal cephalopharyngeal skeleton, led to the identification of the <em>Calliphora</em> spp. fly genus. Treatment consisted of removal of the larvae and surgical debridement, then spray of antibiotic and toxic drug. Following removal of larvae and treatment, the symptoms completely resolved within the last hour and remained asymptomatic several weeks later. This is the first report of wound myiasis in a Flamingo (<em>Phoenicopterus </em><em>ruber)</em> by the facultative myiasis agent <em>Calliphora</em> spp. in Iran and the world.</p> Roghayeh NOROUZI Saeed KAHNAMOOIE Ali OZBANDI ##submission.copyrightStatement## 2018-12-22 2018-12-22 13 4 669 672 Symptomatic Leishmaniasis in an Italian Segugio Dog from Northeastern Romania: A Case Report <p>Leishmaniasis is a zoonotic parasitosis caused by a diphasic protozoan of the genus <em>Leishmania</em>. The dogs are considered the main domestic reservoir of <em>L. infantum</em> and its transmission occurs mainly through sand flies. We report the case of a 10 yr old Italian Segugio dog in Mar 2016 from Iasi County-Moldova Region, northeastern Romania, referred to a private clinic with progressive weight loss, dermal lesions over the muzzle, foot pads and over the right and left tarsal joints. The dog was born in Torino, Italy and transferred to Romania, with a history of regular travelling between these two countries. The physical examination revealed multiple cutaneous lesions with alopecia together with polyarthritis, lymphadenopathies, fatigue and weight loss. Neither fever or nor diarrhea were observed. The serological test (enzyme-linked immunosorbent assay) showed a positive result for Leishmaniasis. Light microscopy of the stained smears prepared from popliteal lymph node puncture failed to identify the amastigotes. The infection was treated using pentavalent antimonial therapy for eight weeks and Allopurinol for eight months. After nine months follow-up the dog presented with an improved body condition and no signs of recurrence.</p> Oana Irina TANASE Constantin DARABAN Elena VELESCU Dragos BOGHEAN Florentina BOCANETI-DARABAN ##submission.copyrightStatement## 2018-12-22 2018-12-22 13 4 673 678 Current Challenges to the Control of Echinococcosis in China <p>Current Challenges to the Control of Echinococcosis in China</p> Yixia CHEN Juntao DING ##submission.copyrightStatement## 2018-12-22 2018-12-22 13 4 679 680 Response to: Comment on “Zoonotic and Non-zoonotic Parasites of Wild Rodents in Turkman Sahra, Northeastern Iran” <p>Response to: Comment on “Zoonotic and Non-zoonotic Parasites of Wild Rodents in Turkman Sahra, Northeastern Iran”</p> Monireh GHOLIPOURY Hamid Reza REZAI Somayeh NAMROODI Fatemeh ARAB KHAZAELI ##submission.copyrightStatement## 2018-12-22 2018-12-22 13 4 681 682