Iranian Journal of Parasitology
Volume:10 Issue: 1, Jan-Mar 2015

Leishmaniasis is a parasitic infection that may lead to a variety of manifestations. In Iran, cutaneous leishmaniasis (CL) has a high prevalence. There are many treatment modalities for CL. The use of oral terbinafine in the treatment of CL has recently been considered. The aim of this study was to compare combination of oral terbinafine plus cryotherapy versus systemic meglu­mine antimoniate plus cryotherapy in CL. Patients with proven direct smear for CL were divided randomly in 2 groups of 40 cases. For the first group systemic glucantime prescribed (IM, 15 mg/kg/day) for 3 weeks. For the second group oral terbinafine as two folds of usual dose in the treatment of fungal diseases prescribed [125 mg/day for body weight (BW) <20 kg, 250 mg/day for BW 20-40 kg, 500 mg/day for BW>40 kg] for 4 weeks. Both groups received cryotherapy every 2 weeks for 4 weeks. The patients were followed monthly for 3 months after the treatment. Partial (HR= 0.55, CI 95%= 0.3-1.1) and complete (HR= 0.53, CI 95%= 0.3-0.98) clinical improvement in terbinafine group was much slower than glucantime group, although at the end of treatment protocols no signifi­cant difference between groups were statistically observed (P=0.27). Considering more convenient suitable route of administration and approximately comparable results, it seems that terbinafine can be used as an alternative treatment, especially in the case of allergy or resistance to systemic glucantime.

Fasciola hepatica and F. gigantica are the causative agents of fasciolosis in domestic animals and humans. Based on the morphometric criteria, differential diagnosis between them is problematic. In addition, intermediate forms of Fasciola have been found in Iran, which makes the differentiation more difficult. The aim of the present study was to provide molecular evidence for the existence of F. gigantica in Iran using sequencing analysis of ND1 and PCR-RFLP analysis of ITS2 regions and to study the intraspecies variations of F. gigantica based on mitochondrial ND1 gene polymorphism. Forty Fasciola spp. samples collected from four distinct provinces (Fars, Khuzestan, Gilan, Khorasan Razavi) in Iran were collected for morphological and molecular characterization. In molecular method, PCR-RFLP analysis of ITS2 us­ing pagI restriction enzyme was used as a screening approach for F. gigantica differentia­tion. Then mitochondrial DNA sequence variations in the ND1 gene were used for phylogenetic analysis. Based on the morphometric criteria and RFLP analysis, 14 parasitic sam­ples were initially identified to be F. gigantica. Phylogenetic results showed that there are at least 10 different genotypes of F. gigantica in Iran, which are different from those existing in the GenBank. Twenty-six points out of 410 base pairs of se­quenced ND1 gene in 10 varieties of F. gigantica were diagnosed to be polymorphic. From 26 points of polymorphism, only eight resulted in the post-translational amino acid changes in ND1 gene product structure. Data revealed noticeable genetic diversity (up to 4.63%) between different varieties of F. gigantica in Iran.

In this study, two-dimensional gel electrophoresis (2-DE) method was applied to determine and compare the protein spots expressed in the two field isolates of Leishmania major and recovered from the patients who were clinically sensitive and resistant to Glucantime® treatment. Leishmania parasites were isolated from the cutaneous lesions of two CL infected patients in Shiraz, south of Iran. The species of the two isolates were identified as L. major using Nested-PCR. Sensitivity (Sh-214S) and resistance (Sh-120R) of the two isolates to meglumine antimonite were checked by the standard in vitro assays. Both sensitive and resistant L. major isolates were harvested in RPMI 1640 medium. Protein extractions were performed using TCA/Acetone method and the protein spots were determined by a two-dimensional gel electrophoresis (2-DE). The gels were stained with silver nitrate and analyzed by Image Master 2D Melanie-6 software. About 2967 protein spots were detected. Overall, 89 protein spots repre­sented considerable changes of expression in the resistant isolate of L. major compared to the sensitive isolate. Of these, 60 and 29 protein spots were up-and down regulated, respectively. In addition, 11 protein spots present in the resistant isolate were noticed to be absent in the sensitive isolate. A number of proteins showed significant changes of expression in the drug-resistant L. major; moreover, the roles of these proteins probably enhanced the parasite resistance to the drug and increased parasite survival in the cells.

Cystic echinococcosis is an important zoonosis caused by the Echinococcus granulosus with a substantial impact in human and animal health in en­demic areas. The purpose of the present study was serodiagnosis optimizing of dog echinococcosis in order to achieve a rapid diagnostic method. Eight dogs were challenged with protoscoleces in order to have posi­tive echinococcosis serum and 2 two-month old puppies were used as uninfected controls. Colloidal gold was prepared by controlled reduction of a boiling solu­tion of chloroauric acid (H [AuCl4]) with sodium citrate and labeled with recombi­nant EPC1. Dot immunogold filtration assay (DIGFA) was developed by coating rEPC1 labeled colloidal gold on nitrocellulose membrane. The canine sera, taken three times including, 15, 28 and 35 days post infection were tested. A total of 30 serum samples including 24 sera from 8 infected dogs and 6 sera from 2 puppies were comparatively detected with both DIGFA and ELISA. Gold labeled antigen, showed a dark purple dot with agglutination parti­cles in positive sera and light purple dot without agglutination in negative sera. Among 30 serum samples, 23 were positive and 7 were negative with DIGFA and 24 were positive, 6 were negative with ELISA. DIGFA as a rapid and simple procedure could be utilized in quickly diagnosis of echinococcosis.

The aim of this study was to detect low parasite and asymptomatic malaria infections by means of three malaria diagnostic tests, in a low transmission region of Minab district, Hormozgan Province, southern Iran. Blood samples of 200 healthy volunteers from Bagh-e-Malek area were evaluated using microscopic, rapid diagnostic tests (RDT) and nested-PCR to in­spect malaria parasite. The results showed no Plasmodium parasite in subjects by means of micros­copy and RDT. However, 3 P. vivax positive samples (1.5%) were discov­ered by Nested-PCR while microscopy and RDT missed the cases. Microscopy as the gold standard method and RDT correctly identi­fied 98.5% of cases, and molecular analysis is sensitive and reliable, especially in the detection of "asymptomatic" infections for active case surveillance. Regarding the existence of asymptomatic malaria in endemic area of Hormozgan, Iran, nested-PCR could be considered as a sensitive tool to interrupt malaria transmis­sion in the country, beside the microscopic and RDT methods.

The trematodes of the genus Fasciola (the liver flukes) are among the well-known instances of food-borne parasites worldwide. Differentiation of Fasciola species is important because of their different transmission and epidemiologi­cal characteristics. The current study was undertaken to discriminate Fasciola species in the domestic ruminants of Urmia city, Iran. Adult flukes were isolated from the naturally infected livers of the slaugh­tered water buffaloes and sheep. The flukes were initially identified based on morphological and morphometric parameters. A 618-bp-long fragment of the 28SrRNA gene of Fasciola was amplified by polymerase chain reaction (PCR). The amplified fragment was digested by DraII or AvaII enzymes for a restriction fragment length polymorphism (RFLP) analysis and sequenced for the phyloge­netic tree construction. Based on the morphometric examination, the flukes belonged to F. hepat­ica, F. gigantica and an intermediate Fasciola form. The PCR-RFLP analysis was able to differentiate F. hepatica from F. gigantica. While the phylogenetic reconstruc­tion justified, to some extent, the morphological diagnosis, it failed to segregate F. hepatica from F. gigantica identified in this and the previous studies. To resolve fully the problem of taxonomy and evolution in Fasciola species, employing a broad range of molecular and morphological approaches is necessary. This is crucial for epidemiological surveys and successful clinical manage­ment of their infection.

Visceral leishmaniasis (VL) as one of the most important human parasitic disease is endemic in some parts of Iran. Several cases of VL have been reported recently in the Ilam Province. The current study aimed to assess the pre­sent status of human VL in the region. A random cluster sampling method was used to collect 456 serums sam­ples from the children up to 12 years of age and 10% of adults living in urban and rural areas of the province. All the collected serum samples were tested by di­rect agglutination test (DAT) to detect anti- Leishmania infantum antibodies. Of the examined 456 serum samples with direct agglutination test (DAT), only 21 (0.43%) sera showed anti- Leishmania antibodies at titers 1:400 and higher. Distribution of anti- Leishmania antibodies titers were: 1:400(n=4), 1:800(n=11), 1:1600(n=3), 1:3200(n=1), and 1:6400(n=1). Individuals with titers ≥1:3200 showed clinical signs and symptoms such as fever and splenomegaly. The highest and lowest seropositivity were observed in the age groups of 5–9 and >15 years old, respectively. There were no significant difference between the rate of seropositiv­ity in males and females. VL with a low prevalence circulates in some parts of Ilam province, particularly in the southern parts. Complementary studies should be needed to find animal reservoir hosts and vectors. Furthermore, health systems and physicians should pay particular attention to the disease.

Adult worms of Orientobilharzia turkestanicum live in the portal veins, or intestinal veins of cattle, sheep, goat and many other mammals caus­ing orientobilharziasis. Orientobilharziasis causes significant economic losses to livestock industry of Iran. However, there is limited information about geno­types of O. turkestanicum in Iran. In this study, 30 isolates of O. turkestanicum obtained from sheep were characterized by sequencing mitochondrial cytochrome c oxidase subunit 1 (cox1) and nicotinamide adenine dinucleotide dehydrogenase subunit 1 (nad1) gene. The mitochondrial cox1 and nad1 DNA were amplified by polymer­ase chain reaction (PCR) and then sequenced and compared with O. turkestanicum and that of other members of the Schistosomatidae available in GenBankTM. Phylogenetic relationships between them were re-constructed using the maximum parsimony method. Phylogenetic analyses done in present study placed O. turkestanicum within the Schistosoma genus, and indicates that O. turkestanicum was phylogenetically closer to the African schistosome group than to the Asian schistosome group. Comparison of nad1 and cox1 sequences of O. turkestanicum ob­tained in this study with corresponding sequences available in GenbankTM re­vealed some sequence variations and provided evidence for presence of microvari­ents in Iran.

Dense granules are immunodominant proteins for the standardiza­tion of immunodiagnostic procedures to detect neosporosis. In the presented study different fragment of a dense-granule protein was evaluated for serodiagnosis of Neospora caninum in cattle and water buffalo. NcGRA7, from N. caninum tachyzoites was amplified. PCR product and pMAL-c2X plasmid were digested with EcoR1 restriction enzyme and ex­pressed in Escherichia coli to evaluate its competence for detection of anti- N. cani­num antibodies with ELISA in comparison with commercial IDEXX ELISA. Further­more, 230 sera of presumably healthy cattle and water buffaloes (108 cattle and 122 water buffaloes) were analyzed by both tests to determine the agreement of these two procedures. Sensitivities and specificities of NcGRA7-based ELISA were 94.64% and 90.38% respectively using sera of cattle, but were 98.57% and 86.54% in the case of buffaloes respectively. A good correlation between the results of IDEXX ELISA and ELISA based on recombinant NcGRA7 for detecting N. caninum antibod­ies was appeared. Analyzing by Mc Nemar´s showed that NcGRA7-based ELISA has acceptable capability to differentiate the positive results in comparison with IDEXX ELISA. NcGRA7-based ELISA considering utilized new fragment of ge­nomic DNA is a good tool for serodiagnosis of anti- N. caninum antibodies for screening and epidemiological purposes on cattle herd and water buffaloes as well.

This study was conducted to explain the trend of Cutaneous Leishmani­asis (CL) in Kermanshah Province, western Iran from 1991- 2012. In this analytical-descriptive study, all of the patients suspected to CL in Kermanshah Province were studied. Smears were prepared from most of them, stained with Giemsa, and examined microscopically for Leishman bodies. A few of the patients were diagnosed according to physician diagnosis and response to glucan­time. Questionnaires were completed for all of them. Collected data were analyzed using SPSS-21 and Spearman and Kendall tests. From 1991 to 2012, 1684 cases of CL were recorded. In the years 2011-12 the frequency of CL in the province reached to 7.4/100,000, which showed a remarkable increase in the frequency of new cases CL in the province. In the years 2011-12 about 47% of total cases of CL in the province, was seen in Ghasr-e-Shirin district,which the frequency of CL in this district reached to 264.5/100,000. Fre­quency of CL in males and females was 57% and 43% respectively. The most cases of CL were seen in housewives (32.1%) and age groups of 20 to 29 years old (19.9%). In addition, the most cases of disease were seen in winter (41.8%) and fall (28.9%). CL is increasing remarkably in warm climates of the province, and Ghasr-e-Shirin must be of particular interest, as the most infected district in the province.

Coccidiosis is a serious protozoal disease of poultry. The identifica­tion of Eimeria species has important implications for diagnosis and control as well as for epidemiology. The molecular characterization of Eimeria species infecting Egyptian baladi chickens was investigated. Eimeria species oocysts were harvested from intestines of naturally infected Egyptian baldi chickens. The morphometry characterization of oocysts along with COCCIMORPH software was done. The DNA was extracted ini­tially by freezing and thawing then the prepared samples was subjected to commer­cial DNA kits. The DNA products were analyzed through conven­tional polymerase chain reaction by using amplified region (SCAR) marker. The PCR results confirmed the presence of 7 Eimeria species in the examined fecal samples of Egyptian baldi breed with their specific ampilicon sizes being E. acervulina (811bp), E. brunette (626bp), E. tenella (539bp), E. max­ima (272bp), E. necatrix (200bp), E. mitis (327bp) and E. praecopx (354bp). A sequencing of the two most predominant species of Eimeria was done, on E. tenella and E. máxima. Analysis of the obtained sequences revealed high identi­ties 99% between Egyptian isolates and the reference one. Similarly, E. maxima isolated from Egyptian baldi chickens showed 98% nucleotide identities with the reference strain. Only single nucleotide substitution was observed among the Egyptian E. tenella isolates (A181G) when compared to the reference one. The Egyptian isolates acquired 4 unique mutations (A68T, C164T, G190A and C227G) in compared with the reference sequence. This is the first time to identify the 7 species of Eimeria from Egyptian baladi chickens.

Neospora caninum is considered one of the major causes of repeated abortions in livestock. This study aimed to determine the seropositivity to N. caninum using indi­rect ELISA and the influence of the infection on the occurrence of abortions in selected dairy herd in Slovakia. Blood samples were obtained from 490 cattle over a period of two years and were tested for N. caninum antibodies using indirect ELISA. The presence of specific antibodies in the herd was detected in 118 (24.1%) cows. According to selected groups; 117 (41.0%) cows with a history of abortion, 65 (43.3%) heifers and 223 (2.2%) cows without abor­tions were tested positive to Neospora. Vertical transmission of N. caninum domi­nated in examined herd and the relative risk (RR) of dam-daughter seropositivity in progenies of seropositive mothers was 2.1 times higher than in progenies of seronega­tive dams. Molecular analyses of aborted foetuses of seropositive mothers showed the presence of Neospora DNA. However, 23 (28.1%) of heifers born to seronegative cows were seropositive, indicating also the postnatal transmission of the infection from the environment. Study revealed significant correlation between the presence of specific antibodies and the occurrence of abortions, the risk of abor­tion in seropositive animals was 3.8 times higher than in seronegative ones. Incorrect farm management contributed to spread and circulation of neosporosis in entire dairy herd what could significantly impair the reproduction and economic parameters of breeding.

Human Echinostomiasis is an intestinal disease caused by the members of family Echinostomatidae parasites. The aim of present research was to identify echinos­tomatidae cercariae emitted by Lymnaea palustris snails from Mazandaran province in the north of Iran based on the morphological and morphometrical charac­teristics of the different stages of experimental parasite life cycle. Echinostomatidae cercariae were collected from L. palustris (Gastropoda: Lymnaeidae) of the north of Iran. To collect metacercaria, 50 healthy snails were infected with cercariae experimentally (50 cercariae for each). To obtain the adult stage, 9 laboratory animals (3 ducks, 2 rats, 2 mice and 2 quails) were fed with 60 metacercaria for each. To identify parasite, the different stages of worm were exam­ined using light microscope and then the figures were draw under camera Lucida microscope and measures were determined. Averagely, 15metacercaria were obtained from each snail that had been previously exposed with cercariae. Ducks presented worm eggs in feces after 10-15 days post-infection. Intestinal worms were collected and identified as Hypoderaeum conoideum on the bases of figures and measures of cephalic collar, the number of collar spine, suckers diameter ratio, testes arrangement, etc. H. conoideum cercariae and adult worm are described. This is the first report of the different stages of the experimental life cycle of this parasite in Iran.

Infection with Trichostrongylus spp. is common among human and herbivorous in most parts of Iran, especially in southern and northern ar­eas. The aim of present study was to identify Trichostrongylus spp. among hu­man population using excreted egg specimens, by the molecular method, in Ma­zandaran Province, northern Iran. Overall, 33 positive fecal specimens were randomly sampled and ex­amined. PCR amplification of ITS2-rDNA region was performed on the iso­lated egg and then a restriction fragment length polymorphism (RFLP) pro­file was considered to discriminate of Trichostrongylus spp. A total of 33 positive fecal specimens, 29(78.9%), 4(12.1%) were found T. colubriformis and T. axei respectively. Our data appear the molecular evi­dence of both human T. colubriformis and T. axei infections in North of Iran. T. colubriformis was the probable most common zoonotic species caus­ing human trichostrongylosis infection in the area.

Acanthocephalans are fish parasites of worldwide distribu­tion, penetrate their thorny proboscis into the intestinal wall of host and absorb nutrients. No diagnostic tool is available except postmortem investiga­tions and identification by parasitologists. The aim of present study was to explore and assign taxonomical status to Pallisentis species preva­lent in food fishes of river Gomti, Lucknow, India. A survey of fishes of river Gomti was carried out during the year 2011-2013. Acanthocephalans recovered from the intestine of Channa punctatus were kept in refrigerator for eversion of proboscis, fixed in A.F.A. fixative (50% alcohol, formalin and acetic acid in ratio of 100: 6: 2.5) for 24 hours further preserved in 70% ethanol. Camera Lucida diagrams of acetoalum carmine stained permanent mounts were made for morphometric studies. Two new species of genus Pallisentis were identified and named as P. channai n. sp. and P. vinodai n. sp., their taxonomical status is based on major characters of proboscis hooks, spines of collar and trunk region, cement gland nuclei. On average 9 fishes were found infected with Pallisen­tis spp. out of 60 fishes examined randomly. Pallisentis spp. are important parasitic infection in Channidae fishes with the prevalence rate of 15%. Two new species of Pallisentis recog­nized from Channa punctatus of river Gomti, Lucknow, India and diagnostic features of genus are given.

To analyze the characteristic performance of magnetic resonance spectroscopy (MRS) and susceptibility weighted imaging (SWI) in cerebral alveo­lar echinococcosis (CAE). We retrospectively analyzed 10 clinical-identified CAE cases MR performance, and summarized the MRS and SWI performance of CAE. The 10 cases of CAE all had the history of primary HAE, among who 6 cases had single lesion (60%), while the rest 4 cases had multiple lesions (40%); and 4 cases were concomitant with lung metastases. MRI performance: T2WI lesions were coal-like low-signal shadow, with multiple small vesicles in­side the lesions; MRS performance: NAA, Cho and Cr significantly reduced, an abnormally high and steep crest was found at 1.4 ppm; the phase diagram and strength diagram of SWI showed isointensity. The MRS and SWI of CAE could provide important supplemental information for the diagnosis of CAE, especially the abnormally high and steep crest at 1.4 ppm provide the reliable image basis for the diagnosis and differen­tial diagnosis of CAE.

A 47-year-old man presented as a dyspnea and fatigue. A transthoracic echocardiog­raphy (TTE) performed to assess valvar heart disease and left ventricu­lar (LV) function showed a large hydatid cysts in the inter ventricular septum bulg­ing to posterior of heart and compressing its overlying muscle as whitish round mass. The patient underwent elective surgery by evacuation of septal hydatid cysts from posterior inter ventricular aspect of septum and capitonnage of cavity. His postoperative course was uneventful. Post-operative albendazole for prophylaxis of procedures were recommended. The patient was scheduled for regular follow-up, to check for any recurrences or late complications.

Hydatid cyst is an important endemic zoonosis in Iran. It may be seen in various organs of body. Musculoskeletal system is rarely involved by hydatid cyst, the larval form of Echinococcus granulosus. On clinical basis, it may resemble any soft tissue tu­mor. A 70-years old housewife living in Ardoghesh, a village in Neyshabur, north­east Iran, was admitted to general surgery clinic because of a painless mass in the back of her left thigh. This case emphasizes that hydatidosis should be included in differential diagnosis of any soft tissue mass especially in regions where hydatidosis is endemic.

A 40-year-old male patient presented to us with complaints of pain in abdomen for the past 2 weeks and fever for 3 days. The patient gave history of being previously operated for hydatid cyst of liver 15 years back. His chest radiograph and com­puted tomography scan revealed a cystic lesion in the right lobe of liver and a cyst in left lung. The patient was managed surgically. Aspirate from the cyst fluid showed plenty of hooklets and scolices of Echinococcus granulosus. An intact brood capsule was also seen. Diagnosis of hydatidosis was further confirmed by histopatho­logical examination. Post-operative the patient had a good recovery.

Strongyloides stercoralis (SS) is a unique nematode with an auto infective cycle, so that it completes its life cycle within the human host and can live there for many years. In immunocompromised patients, infection can cause Strongyloides hyperinfection syn­drome (S.H.S) that is associated with serious morbidity and mortality. As various infections are one of the leading causes of membranoproliferative glomerulonephri­tis (MPGN), we should consider subclinical strongyloidiasis as a possible underlying disease, especially in endemic areas. Here we describe a case of strongyloidiasis following immunosuppressive therapy for MPGN, the diagnosis of which was made, only a few hours before death, by stomach biopsy.