فهرست مطالب

Iranian Journal of Parasitology
Volume:13 Issue: 4, Oct-Dec 2018

  • تاریخ انتشار: 1397/10/15
  • تعداد عناوین: 25
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  • Hamid DANESHVAR, Amir TAVAKOLI KARESHK, Iraj SHARIFI, Alireza KEYHANI, Arash ASADI, Razieh TAVAKOLI OLIAEE Pages 515-523
    Background
    Different background of immunity responses determine resistance or susceptibility of certain mouse strains to Leishmania major 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 L. major, they have been less studied in Leishmania fields. The aim of present study was to determine whether these components play a role in the protection of skin against Leishmania infections.
    Methods
    The animal model of Leishmania infection was established by the subcutaneous inoculation of 5×106(parasites/ml) from the stationary phase of L. major 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.
    Results
    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 L. Major parasites. We also observed a significant up-regulation of all aforementioned AMPs genes in BALB/c mice at selected times compared to another strain.
    Conclusion
    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.
    Keywords: Antimicrobial peptides, Cathelicidin, Cathelin-related antimicrobial peptide, Cytokine, Defensin, Leishmania major, Mouse ?-defensin
  • KobraKobra PIRI, Massoud SAIDIJAM, Amirhossein MAGHSOOD, Mohammad MATINI, Mohammad FALLAH Pages 524-531
    Background
    Fascioliasis is a common disease among humans and animals. Having global distribution, disease is developed by hepatic trematodes, Fasciola hepatica and F. gigantica. The main objective of this research was determining the prevalence of Fasciola species in Hamadan livestock and identifying those using PCR-RFLP.
    Methods
    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 Fasciola (41 worms from sheep, 22 worms from goats and 12 worms from cattle) were examined by PCR-RFLP method.
    Results
    Totally, 100 livers were infected to Fasciola species (total prevalence: 0.74%; sheep 0.5%, goats 1.4%, cattle 1.5%). In the molecular results, prevalence of F. hepatica was higher than (92.5% in the sheep) F. gigantic, and in the cattle 91.5% and in the goats was 54.5% F. hepatica. Genotyping identified species confirmed intermediated types as F. hepatica.
    Conclusion
    Prevalence of Fasciola in this province is not so high. Intermediate types identified by PCR-RFLP method determined as F. hepatica by nucleotide sequencing. Because of morphological differences and interspecies variety, the accurate identification of Fasciola species needs using nucleotide sequencing.
    Keywords: Prevalence, Fasciola, PCR-RFLP, Iran
  • Nagwa SHABAN ALY, Ibrahim BAYOUMI, Rabab SELEM, Manal KARDOUSH, Gehan RASHED, Ahlam MOHARAM Pages 532-540
    Background
    The traditional basis of diagnosis is identification of Giardia lamblia 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 Giardia cysts may facilitate diagnosis of G. lamblia in stool specimens.
    Methods
    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 G. lamblia cysts antigen was prepared by two-phase sucrose gradient technique. Polyclonal antibody against purified G. lamblia 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.
    Results
    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%.
    Conclusion
    Diagnosis of G. lamblia 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.
    Keywords: Giardia lamblia, Dot ELISA, Nano technology
  • 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 Pages 541-548
    Background
    Visceral Leishmaniasis (VL) caused by protozoa belonging to the genus Leishmania, 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).
    Methods
    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.
    Results
    The mean level of IL-10 in DATposLOW of asymptomatic VL and PKDL was significantly higher than endemic healthy (P<0.05). The cytokine polarization index (IFN-γ versus IL-10) was significantly low in PKDL cases compared with asymptomatic VL cases in DATposLOW titre (P<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.
    Conclusion
    Collectively we surmise from our data that asymptomatic VL can also play an important role like PKDL in transmission of VL.
    Keywords: Visceral leishmaniasis, Asymptomatic infection, Cytokine, Direct agglutination test, Interleukin-10, IFN-gamma, Tumor necrosis factor-alpha
  • Hossein DEHGHAN, Mohammad Ali OSHAGHI, Seyed Hassan MOSA, KAZEMI, Mohammad Reza ABAI, Fatemeh RAFIE, Mehdi NATEGHPOUR, Habib MOHAMMADZADEH, Leila FARIVAR, Mulood MOHAMMADI BAVANI Pages 549-555
    Background
    Plasmodium berghei 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 Anopheles stephensi 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 P.berghei–A.stephensi-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.
    Keywords: Plasmodium berghei, Anopheles stephensi, BALB, c, malaria, life cycle
  • Berhanu ELFU FELEKE Pages 560-566
    Background
    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.
    Methods
    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.
    Results
    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.
    Conclusion
    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.
    Keywords: Prevalence, Hookworm, Children, Urban, Rural, Ethiopia
  • AbdolhosseinAbdolhossein DALIMI, Anita MOHAMMADIHA, Mehdi MOHEBALI, Asad MIRZAEI, Mohammadreza MAHMOUDI Pages 567-576
    Background
    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.
    Methods
    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.
    Results
    The cause of all 70 VL cases, were L. infantum, so, the dominant specie is L. infantum. The sequencing results of all VL cases and RFLP analysis corroborate each other. Discrimination of Iranian Leishmania 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 Leishmania spp. DNA was confirmed by amplification of variable area of the minicircle kDNA (conserved sequence blocks (CSB)).
    Conclusion
    Low divergence and high likelihood were seen among L. infantum 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.
    Keywords: Phylogeny, Leishmania infantum, PCR-RFLP, Sequencing, Iran
  • Mitra SALEHI, Saeed YAGHFOORI, Pejman BAHARI, Mohsen SEYEDABADI, Sima PARANDE SHIRVAN Pages 577-586
    Background: Echinococcus granulosus 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 E. granulosus from domestic animals in nine districts of North Khorasan Province using the mitochondrial cox1 gene sequence.
    Methods
    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 E. granulosus genotypes from various locations.
    Results
    Sequencing of the amplified products revealed the presence of G1 as dominant genotype, G3 and Echinococcus canadenesis 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.
    Conclusion
    This study provided knowledge on the identity of E. granulosus 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.
    Keywords: Cox1 gene, Genotypes, Hydatid cyst, Molecular epidemiology, Iran
  • Mahmoud RAHDAR, Abdollah RAFIEI, Rohollah VALIPOUR NOROUZI Pages 587-593
    Background
    We aimed to use of IL-12 and IFN-γ for prevention and treatment of hydatidosis in experimental animal models.
    Methods
    This experimental study was conducted in Ahvaz City, southwest of Iran in 2017.  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.
    Results
    There issignificant reduction in number, size, and weight of cysts in test groups.
    Conclusion
    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.
    Keywords: Hydatid cyst, Cytokine therapy, Mice
  • Tams SULI, Gordana KOZODEROVI, Aleksandar POTKONJAK, Stanislav SIMIN, Verica SIMIN, Vesna LALOEVI Pages 594-601
    Background: Blastocystis is a common protist colonizing the gastrointestinal tract of humans and various animals. Pigs have been suggested to be a reservoir for human Blastocystis 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 Blastocystis sp. in pigs.
    Methods
    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.
    Results
    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%.
    Conclusion
    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.
    Keywords: Blastocystis, Pig, PCR, In vitro culture, Sensitivity
  • Wen, Chao LI, Kai WANG, Yan LI, Li, Ping ZHAO, Yi XIAO, You, Fang GU Pages 602-610
     Background: In pigs, several different trichomonad species such as Tritrichomonas foetus, Tetratrichomonas buttreyi and Pentatrichomonas hominis 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.
    Methods
    The prevalence of T. suis, T. buttreyi and P. hominis 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.
    Results
    The prevalence rates for T. suis, T. buttreyi and P. hominis 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 T. suis and T. buttreyi in nursery pigs and P. hominis in preweaned pigs. The SSU rRNA sequences from T. suis and P. hominis showed 100 % homology with their respective homologous database sequences. However, we observed minor allelic variations in the SSU rRNA sequences from T. buttreyi, 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.
    Conclusions
    These findings highlight the potential risk posed by pigs in the transmission of trichomonad infections to humans and other animals.
    Keywords: Tritrichomonas suis, Tetratrichomonas buttreyi, Pentatrichomonas hominis, Pigs, China
  • Hamed KIANI, Aliyar MIRZAPOUR, Iraj MOBEDI, Adel SPOTIN, Seyyed Javad SEYYED TABAEI, Mohsen RAHIMI Pages 611-617
    Background
    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.
    Methods
    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).
    Results
    65.7% (23/35) of zoo animal species were infected with intestinal parasites. The superfamily Trichostrongyloidea (6/16) and Strongylus sp. (16/4) were the most prevalent helminthic infections, while Blastocystis sp. (6/14), Entamoeba cyst (3/14) and Eimeria sp. (3/14) were the common protozoan parasites. For the first time, Bivitellobilharzia nairi egg was identified an elephant at Iran. Intestinal parasitic infections were apparently circulating among animals of the Eram park zoo.
    Conclusion
    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.
    Keywords: Intestinal parasites, Morphometric criteria, Zoo animal, Bivitellobilharzia nairi, Iran
  • Biswa RANJAN MAHARANA, Anup KUMAR TEWARI, Naduvanahalli RAJANNA SUDHAKAR, Chinmoy MISHRA Pages 618-626
    Background
    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.
    Methods
    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 E. coli. 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.
    Results
    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 in silico Te-PFR1 (T. evansi PFR1) nucleotides sequence analysis revealed its close homology with the other members of the order Kinetoplastida.
    Conclusion
    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 Trypanosoma species.
    Keywords: Para flagellar rod pro-tein 1, SDS-PAGE, Trypanosoma evansi, Western blot
  • zgür Yaar CELK, Burçak ASLAN CELK Pages 627-631
    Background
    Fasciolosis is a disease of the liver caused by trematodes in the family of Fasciolidae, particularly by Fasciola hepatica and Fasciola gigantica. The aim of this study was to investigate the prevalence of F. hepatica in sheep using the ELISA method, and in hair goats by post-mortem liver examination in the Siirt region, Turkey.
    Methods
    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 F. hepatica test). In order to determine the prevalence of fasciolosis in hair goats, 580 slaughtered goats were examined for F. hepatica by incisions in the liver, gallbladder, and bile ducts.
    Results
    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.
    Conclusion
    F. hepatica 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.
    Keywords: Sheep, Hair Goat, Fasciola hepatica, Turkey
  • Jorge, Luis DE, LA, ROSA, ARANA, Raquel TAPIA, ROMERO Pages 632-636
    Background
    The dog, Canis familiaris, 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.
    Methods
    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).
    Results
    Eggs of Toxocara canis (41%), Ancylostoma caninum (8%) and Dipylidium caninum (3%) were found in the rural environment but none in the urban zone. A frequency of 19% of Toxocara 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 Toxocara were found in 33% samples of puppies with history of antiparasitic treatment.
    Conclusion
    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.
    Keywords: Helminth, Puppy, Urban-environment, Rural-environment
  • Maryam KHALILI, Mahmoud MAHAMI, OSKOUEI, Abbas SHAHBAZI, Abdolrasoul SAFAIYAN, Nader MOHAMMADZADEH, GHESHLAGHI, Leyla MAHAMI Pages 637-642
    Background
    This study investigated the presence of specific antibodies against Toxoplasma gondii infection among people with diabetes (type I and II) in comparison with control group.
    Methods
    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 T. gondii were measured using the chemiluminescence immunoassay (CLIA) method. Chi-square and One-Way ANCOVA were used for data analysis.
    Results
    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- T. gondii IgM respectively. Anti- T. gondii 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-T. gondii antibodies in type I and II diabetes and non-diabetic groups.
    Conclusion
    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.
    Keywords: Diabetes mellitus, Toxoplasma gondii, Chemiluminescence, Immunoassay
  • Hatem SHALABY, Amira EL NAMAKY, Omnia KANDIL, Noha HASSAN Pages 643-647
    Background
    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 Balanites aegyptiaca fruits extract on the cuticle of adult worm of Toxocara canis as naturally alternative therapy.
    Methods
    B. aegyptiaca 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 B. aegyptiaca fruits on adult T. canis after 24 and 48 h incubating the parasites in Ringer solution containing 240 μg/ml Balanites extract was determined by scanning electron microscopy.
    Results
    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 Toxocara worms from dog’s intestine.
    Conclusion
    The use of this plant offers a chance for new nematocidal agent, which is economical alternative for the more expensive anthelmintics.
    Keywords: Toxocara canis, Balanites aegyptiaca fruits, In vitro effect, Cuticle, Scanning electron mi-croscopy
  • Emad AHMADIARA, Seyed Hossein HOSSEINI, Fatemeh JALOUSIAN, Hossein Ali EBRAHIMZADEH MOUSAVI Pages 648-654
    Background
    The position of Digramma interrupta remains disputable as it was raised by Cholodkovsky from Ligula alternans. This study aimed to survey the evolutionary relationships and the taxonomic position of D. interrupta and L. intestinalis. It also intended to support or reject the validity of D. interrupt as an independent genus and its correlation with L. intestinalis on the basis of their morphological characteristics and a study on molecular data.
    Methods
    Overall, 1301 fish varieties, including 883 Alburnoides bipunctatus and 418 Abramis brama, were collected from north and north-western parts of Iran. A. bipunctatus samples were obtained from fresh water sources of the Maragheh dam (northwest) and the Ramesar Lake (north). Moreover, samples of A. brama 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.
    Results
    Nucleotide variation between L. intestinalis and D. interrupta samples amounts to about 3% to 7%. Between samples of L. intestinalis and GenBank data, and also between D. interrupta specimens and GenBank data, the diversity was seen for about 1% to 3%. Moreover, about 1% to 4% nucleotide variation was seen only in L. intestinalis samples caught from the same host, which could be supplementary to the presence of a species and/or strains in this genus.
    Conclusion
    Maybe D. interrupta was just a rare diplogonadic form of the Ligula species, not a different genus and not synonymous with the Ligula genus, but only another species of the Ligula genus.
    Keywords: Ligula intestinalis, Digramma interrupta, Morphology, Molecular diagnosis, Iran
  • De Godoy NATALIA SOUZA DE, Aiello VERA DEMARCHI, de Souza REGINA MAIA, Okay THELMA, Braz LUCIA MARIA ALMEIDA Pages 655-660
    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 andRV1/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 comorbiditieswho 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.
    Keywords: Visceral leishmaniasis, HIV coinfeccion, Mucocutaneous manifestation, Leishmania (L.) infantum chagasi, RFLP
  • Hajar ZIAEI HEZARJARIBI, Mahdi FAKHAR, Rezvan YALVEH, Sayed Abdollah MOUSAVI, Elham Sadat BANIMOSTAFAVI Pages 661-663
     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.
    Keywords: Children, Vulvar bleeding, Leech
  • Davood YADEGARINIA, Shabnam TEHRANI, Zahra DOOSTI Pages 664-668
    Fascioliasis is a zoonotic disease caused by liver flukes of the genus Fasciola, as F. hepatica, and F. gigantica, 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 Fasciola, 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.
    Keywords: Fascioliasis, Cholangiocarcinoma, Endoscopic retrograde cholangiopancreatog-raphy
  • Roghayeh NOROUZI, Saeed KAHNAMOOIE, Ali OZBANDI Pages 669-672
    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 Calliphora spp. in a Flamingo (Phoenicopterus ruber) 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 Calliphora 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 (Phoenicopterus ruber) by the facultative myiasis agent Calliphora spp. in Iran and the world.
    Keywords: Wound myiasis, Flamingo (Phoenicopterus ruber), Calliphora spp., Northwest Iran
  • Oana Irina TANASE, Constantin DARABAN, Elena VELESCU, Dragos BOGHEAN, Florentina BOCANETI, DARABAN Pages 673-678
    Leishmaniasis is a zoonotic parasitosis caused by a diphasic protozoan of the genus Leishmania. The dogs are considered the main domestic reservoir of L. infantum 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.
    Keywords: Leishmaniasis, Segugio, Serology
  • Yixia CHEN, Juntao DING Pages 679-680
  • Monireh GHOLIPOURY, Hamid Reza REZAI, Somayeh NAMROODI, Fatemeh ARAB KHAZAELI Pages 681-682