Serological study of Human Fasciolosis in Patients Referring to the School of Public Health,Tehran University of Medical Sciences,Tehran, Iran during 2008–2014

Background:

Fascioliasis is a zoonotic disease of livestock and human caused by Fasciola species. Here in, the results of serological evaluation of fascioliasis in people referring to the School of Public Health, Tehran University of Medical Sciences during 2008–2014 are presented.

Methods:

Demographic characterizations, symptoms and eosinophil rate were registered for every patient. Using somatic antigen of Fasciola, ELISA was performed and the results were analyzed. Data of questioners were analyzed as well.

Results:

Among 206 applicants, 24.8% were seropositive for fascioliasis, included 21% female and 28.3% male. Mean range of age of patients was between 13 to 67 yr. The highest rate of seropositivity was found among 20–30 yr old patients. Most of the patients had hypereosinophilia. All patients had history of eating raw vegetables, or drinking unsafe water. Patients were referring from different provinces of Iran, including Gilan, Mazandaran, Tehran, Ardabil, Khuzestan, Lorestan, North Khorasan, Kermanshah, Azerbaijan, Fars, Kordestan, Hamedan and Markazi.

Conclusion:

During recent years, variety of provinces in Iran, where patients with fascioliasis are referred, has been increased. Patients coming from Gilan and Mazandaran provinces were referred early after the onset of their symptoms. Most probably, physicians in Gilan and Mazandaran are more alert on fascioliasis than other provinces. Previous wrong diagnosis was more common among patients referring from other provinces than Gilan and Mazandaran provinces.     

Superoxide Dismutase (SOD) Enzyme Activity Assay in Fasciola spp. Parasites and Liver Tissue Extract

Background

The purpose of this comparative study was to detect superoxide dismutase (SOD) activities in Fasciola hepatica, F. gigantica parasites, infected and healthy liver tissues in order to determine of species effects and liver infection on SODs activity level.

Methods

Fasciola spp. parasites and sheep liver tissues (healthy and infected liver tissues), 10 samples for each, were collected, homogenized and investigated for protein measurement, protein detection and SOD enzyme activity assay. Protein concentration was measured by Bradford method and SODs band protein was detected on SDS-PAGE. SODs activity was determined by iodonitrotetrazolium chloride, INT, and xanthine substrates. Independent samples t-test was conducted for analysis of SODs activities difference.

Results

Protein concentration means were detected for F. hepatica 1.3 mg/ ml, F. gigantica 2.9 mg/ml, healthy liver tissue 5.5 mg/ml and infected liver tissue 1.6 mg/ml (with similar weight sample mass). Specific enzyme activities in the samples were obtained 0.58, 0.57, 0.51, 1.43 U/mg for F. hepatica, F. gigantica, healthy liver and infected liver respectively. Gel electrophoresis of Fasciola spp. and sheep liver tissue extracts revealed a band protein with MW of 60 kDa. The statistical analysis revealed significant difference between SOD activities of Fasciola species and also between SOD activity of liver tissues (P<.05).

Conclusion

Fasciola species and liver infection are effective causes on SOD enzyme activity level.     

Bioaccumulation of Some Heavy Metals in the Liver Flukes Fasciola hepatica and F. gigantica

Background

Heavy metals tend to bioaccumulate in living organisms, and their accumulation has been a major concern. As mammals are known to excrete heavy metals via their bile, it seems to be very promising to analyse metal burdens of parasites that infect the biliary tree such as liver flukes of the genus Fasciola. The present study was carried out to evaluate F. hepatica and F. gigantica as bioaccumulators of heavy metals, and to estimate their use as sensitive markers of environmental pollution with heavy metals.

Methods

A total of 36 slaughtered buffaloes (26 infected and 10 controls) collected from the slaughter-house of Tanta City, Egypt were used. Samples of muscle and liver tissues were taken from each buffalo. A total of 44 adult Fasciola flukes were collected from the 26 infected buffaloes. Quantification of some heavy metals (Cd, Cr, Cu, Pb and Zn) in samples was carried out using electrothermal atomic absorption spectrometry.

Results

Results revealed different concentrations of heavy metals in different host tissues. The adult flukes were classified into F. hepatica (n = 25) and F. gigantica (n = 19). The bio-concentration factor (BCF) of Cr was significantly higher in F. hepatica (P = 0.0465) while BCF of Zn was significantly higher in F. gigantica (P = 0.0189). A comparative study between the two species as regards the BCF was never done before.

Conclusion

The obtained results indicate the possibility of use of Fasciola flukes as markers of environmental pollution with some heavy metals.     

Molecular Identification and Differentiation of Fasciola Isolates Using PCR- RFLP Method Based on Internal Transcribed Spacer (ITS1, 5.8S rDNA,ITS2)

Background

In this study, we used both ITS1 and ITS2 for molecular identification of Fasciola species.

Methods

The region between 18S and 28S of ribosomal DNA was used in PCR-RFLP method for molecular identification of Fasciola species. Ninety trematodes of Fasciola were collected during abattoir inspection from livers of naturally infected sheep and cattle from Khorasan, East Azerbaijan, and Fars provinces in Iran. After DNA extraction, PCR was performed to amplify region ITS1, 5.8S rDNA, ITS2. To select a suitable restriction enzyme, we sequenced and analyzed the PCR products of F. hepatica and F. gigantica samples from sheep and cattle. Tsp509I fast digest restriction enzyme was selected for RFLP method that caused the separation specifically of Fasciola species.

Results

The fragment approximately 1000bp in all of the Fasciola samples was amplified and then digested with the Tsp509I restriction endonuclease. Seventy F. hepatica and 20 F. gigantica were identified of total 90 Fasciola isolates.

Conclusion

The new PCR-RFLP assay using Tsp509I restriction enzyme provides a simple, practical, fast, low cost, and reliable method for identification and differentiation of Fasciola isolates.     

Helminth Parasites of Eastern European Hedgehog (Erinaceus concolor) in Northern Iran

Background

Recently there is a high tendency among exotic pet owners for keeping hedgehogs. This mammal can transfer some significant zoonotic pathogens to human. Hence, the present study was conducted for the first time to prepare a list of helminth parasites of hedgehogs (Erinaceus concolor) in North of Iran.

Methods

Ten (four males and six females) road killed hedgehogs were collected during April to January 2011 in rural areas of Babol city, Mazandaran province, Iran. All of internal organs were scrutinized for helminth burden. The extracted specimens were fixed and preserved in 70% ethanol and then cleared in Lacto-phenol solution. Helminth identification was carried out according to available systematic keys.

Results

All the examined hedgehogs (100%) were infected with parasitic helminth as following: two hedgehogs (20%) were infected with Crenosoma striatum, four hedgehogs (40%) harbored Physaloptera clausa, one (10%) host had Hymenolepis erinacei and three (30%) of them were infected with Nephridiacanthus major.

Conclusion

This is noteworthy that the current survey is the first report of helminth parasites fauna of Eastern European Hedgehog in Iran. Since, this is the first such investigation in our country, more researches are required to perform on unexplored areas of Iran in order to increase our knowledge regarding hedgehog parasitic diseases     

Detection of Coproantigens by Sandwich ELISA in Rabbits Experimentally Infected with Fasciola gigantica

Background

The study was targeted to report the appearance of coproantigens in feces and circulating antibodies in the serum of Fasciola gigantica experimentally infected rabbits.

Methods

Copro Hyper Immune Serum (HIS) and Excretory-Secretory Hyper Immune Serum (ES HIS) antigens were used in a sandwich ELISA for the detection of F. gigantica antigens in feces of 12 rabbits experimentally infected with different doses of F. gigantica encysted metacercariae (EMC) (10, 25 and 30 EMC). The relation between time of appearance of coproantigens in feces and anti-Fasciola antibodies in serum was evaluated.

Results

The earliest diagnostic coproantigen was recorded at 21^st, 25^th and 28^th day post-infection (p.i.) in groups of rabbits infected with 30, 25 and 10 F. gigantica EMC respectively. Both HIS and ES HIS were able to detect coproantigens in feces of rabbits infected with 30 EMC at day 21 p.i. The appearance of F. gigantica coproantigens in feces of infected rabbits was concurrent to the appearance of anti-Fasciola antibodies in blood (3^rd week p.i.). However, coproantigen has specific ability for direct assessment of active infection with minimal cross-reaction with other heterologous parasitic infections.

Conclusion

The findings hold promise for a more accurate diagnostic technique in the near future for suspected Fasciola infection.     

Application of the Reverse Line Blot Assay for the Molecular Detection of Theileria and Babesia sp. in Sheep and Goat Blood Samples from Pakistan

Background

The present study was designed to detect the presence of tick-borne parasites (Theileria and Babesia spp.) in 196 blood samples collected from apparently healthy sheep and goats from two provinces, Punjab and Khyber Pukhtoon Khwa, in Pakistan.

Methods

Reverse line blot (RLB) assay was applied for the parasitic detection by the amplification of hypervariable V4 region of the 18S ribosomal RNA (rRNA) gene. A membrane with covalently linked generic and species specific oligonucleotide probes was used for the hybridization of amplified PCR products.

Results

Parasites were detected in 16% of the ruminant blood samples under study. Two Theileria species, T. lestoquardi and T. ovis, were identified in samples. 25, of the total 32, infected animals were from Khyber Pukhtoon Khwa.

Conclusion

Sheep were more prone to tick borne haemoprotozans as 81% infected samples were sheep as compared to 19% goats (P > 0.001). Risk factor analysis revealed that male (P = 0.03), animals infested by ticks (P = 0.03) and herd composed of sheep only (P = 0.001) were more infected by blood parasites.     

Carbohydrate Detection and Lectin Isolation from Tegumental Tissue of Fasciola hepatica

Background

Fascioliasis is a chronic hepatic disease and may be resulted from mechanical/molecular parasite adhesion to host liver tissue. The aim of this study was to detect surface carbohydrate and lectin, carbohydrate-binding protein isolation that might be responsible of this molecular binding.

Methods

The present experimental work was conducted in the Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran. Fasciola hepatica parasites were collected from abattoir (Saman, Tehran, Iran) and surface mannose-carbohydrate was detected by fluorescein isothiocyanate (FITC) conjugated lectin (Lentil). Lectin of tegumental tissue from F. hepatica was isolated by affinity chromatography and detected by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE).

Results

Mannose carbohydrate was observed on the surface of tegumental tissue from parasite under fluorescence microscope. Carbohydrate-binding protein or lectin with MW of 50 kDa also was isolated from homogenized tegument of helminth.

Conclusion

These results are important for understanding of molecular pathogenesis of F. hepatica at the chronic phase of fascioliasis     

Identification and Genetic Variation of Fasciola Species from Tabriz,North- Western Iran

Background

Fascioliasis is considered as the most important helminthic infection of cattle and sheep. Traditional approaches using morphological and biologic characters cannot cause a certainty in the accurate and precise identification and intra-specific differences of Fasciola spp. In this study, we identified Fasciola species using ITS-1 marker and described genetic variation of each species of the parasite in isolates from Tabriz slaughterhouse in West Azerbaijan Province, north- western Iran.

Methods

Overall, 100 samples (50 from sheep and 50 from cattle) morphologically detected as Fasciola worms were studied for identification of Fasciola species by PCR-RFLP method and intra-species variation of the parasite using RAPD-PCR technique.

Results

A region of approximately 460bp in all samples was successfully amplified. There were no identifiable variations among the size of PCR products. Two and three fragments in samples correspond to F. hepatica and F. gigantica was seen, respectively, through PCR-RFLP method. No difference was seen in digestion pattern according to host (sheep or cattle). Different types of each species of the parasite was observed using RAPD-PCR technique.

Conclusion

We could have an estimate of frequency of F. hepatica and F. gigantic and different genotypes of the parasite in isolates from one locality in north- western of Iran. By extension of such studies in future to other animal hosts (buffalo and goat) and including more regions to sampling, the reliability of the results and their application for control programs in zoonotic diseases will be increased.     

Parasitic Appendicitis From Past to Present in Turkey

Background

Understanding the etiology of appendicitis is important for developing effective treatments the relationship between parasitic appendicitis and various socio-cultural factors were examined, particularly with respect to the incidence of literacy. The aim of the article was to research the relations between parasitic appendicitis and literacy ratio in population.

Methods

Cases of parasitic appendicitis resulting in surgery performed at Buca Seyfi Demirsoy Large State Hospital Surgery Clinic between 2002 and 2009 were retrospectively reviewed and classified according to age, sex, type of parasite, morbidity, and mortality. Studies conducted in different regions of Turkey as well as in other countries were reviewed to determine if there was a relationship between parasitic appendicitis and literacy.

Results

Of the 1,969 appendectomy cases reviewed, nine were classified as parasitic appendicitis (0.45%). Enterobius vermicularis was observed in seven cases and Taenia spp. in two. The average age was 26.4 yr. No morbidity or mortality was found.

Conclusion

The data were compared with a retrospective review of studies conducted in the same regions and a decrease in the rate of parasitic appendicitis was observed during the period between the two reviews. It was determined that a low literacy rate was associated with an increase in the incidence of parasitic appendicitis. Observations made between different countries also produced similar results. In countries where the incidence of parasitic appendiciticis was greater than 1.5%, the literacy rate was less than 88%. To avoid appendectomy resulting from parasites, it is important to increase education and literacy. In some areas, individuals with appendicitis undergo surgery due to a lack of education or poor literacy.     

Molecular Differentiation of Fasciola Species and Characterization of Genetic Diversity of F. gigantica Using NADH Dehydrogenase I (ND1) Gene in the Endemic Areas of Iran

Background:

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.

Methods:

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.

Results:

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.

Conclusion:

Data revealed noticeable genetic diversity (up to 4.63%) between different varieties of F. gigantica in Iran.     

Survey of Migratory Birds (Anatidae: Anas platyrhynchos) for Schistosome Parasites from Mazandaran Province,Northern Iran

Background

The aim of the present study was to survey birds’ schistosomes in migratory birds (Anatidae: Anas platyrhynchos) which are the source of the disease in Mazandaran Province, Northern Iran.

Methods

A number of mallards were bought from the markets of hunted birds. The respiratory tracts (nasal mucosa) and intestinal blood vessels were studied for adult worms. The nasal mucosa was separated and observed by a microscope. In order to separate the visceral schistosomes, after separating intestine, vessel mesenteric was studied under the lamp light and then in saline. The parasite sample was collected for subsequent observation.

Results

Fifteen (13.6%) cases out of 110 studied birds had nasal mucosa contaminated with Trichobilharzia sp. egg. Besides that, two birds had adult worms schistosome visceral i.e. Bilharziella sp.

Conclusion

The elements that cause cercarial dermatitis in aforementioned region are Trichobilharzia sp. and Bilharziella sp. parasites. Thus, it is necessary for the authorities of health, environmental and agricultural organization of the province to cooperate in order to control this disease.     

Identification of Leishmania Species Using PCR Assay on Giemsa-Stained Slides Prepared From Cutaneous Leishmaniasis Patients

Background

Leishmaniasis is a group of diseases that are created by intracellular parasites of Leishmania. Cutaneous leishmaniasis is considered as one of the health problems in some provinces of Iran.

Methods

In this study, a total of 178 Giemsa-stained slides from confirmed cases of cutaneous leishmaniasis were examined. The slides were prepared from the patients with cutaneous leishmaniasis that referred to health centers and infected during the epidemic of cutaneous leishmaniasis in Poldokhtar city, Lorestan Province, Iran in 2006.Genomic DNA from each slide was extracted. After DNA extraction, ITS-PCR was used.

Results

Out of 178 slides, 129 (72.47%) samples had a band in the range of 485 bp and 49 (27.53%) samples 626 bp that matched L. tropica and L. major standard samples, respectively.

Conclusion

This study showed that Leishmania DNA could be efficiently extracted and amplified even from old Giemsa-stained microscopic slides that were stored more than 6 yr. In this study was shown that both L. tropica and L. major species exist in Lorestan Province.     

Mini-Column for Cytoadherence: A New Method for Measuring the Relative Size of Binding Subpopulations in Plasmodium falciparum Isolates

Background

Cytoadherence of Plasmodium falciparum- infected red blood cells to endothelial cells is an important mechanism for parasite survival and a major trigger for diseases pathology. Here, we describe a new adhesion assay in which different cell types (CHO, CHO/CD36 and CHO/ICAM-1) are attached to Cytodex beads in a mini-column format to measure the relative sizes of various binding subpopulations as a percentage of the total population.

Methods

Relative size of CD36 and ICAM-1-binding subpopulations of erythrocytes infected with P. falciparum were measured by amount of parasitemia before and after passing the infected erythrocytes through a particular column.

Results

The mini-column adhesion assay was a suitable method as parasitemia always reduced after passing through a particular column in independent experiments. For example, in a typical experiment using P. falciparum ITG line, 75% of the parasites are retained on a CHO/ICAM-1 while 0% of clone 3D7 is retained.

Conclusion

This work introduced and validated a method for measuring the relative size of parasite binding subpopulations and the selection of them. Also, the mini-column method is of value for assessments of cytoadherence and can be used as tool for different applications.     

Species Identification of Birds Nasal Trichobilharzia in Sari,North of Iran

Background

Cercarial dermatitis is known as an endemic parasitic disease in North of Iran, a hypersensitive skin reaction to the penetration of nonhuman schistosome larvae into human skin. In recent studies in this region, final and intermediate hosts were recognized and Trichobilharzia was identified as the main causative agent of cercarial dermatitis in this region, but to date the parasite species haven''t been identified. Therefore this study was performed to species identification of nasal Trichobilharzia in infected birds for the first time.

Methods

A total of 45 Anas clypeata birds identified as final host, were collected from Sari in North of Iran and infected nasal tissues analyzed using molecular techniques. Genomic DNA was isolated by phenol/chloroform extraction method and ITS region of rDNA were amplified with specific primers its5Trem and its4Trem, then sequenced area were compared with existing records in GenBank.

Results

Twelve samples were infected with Trichobilharzia and results of PCR reaction indicated that all of them belonged to T. regenti. The sequence alignment of present work isolates and those deposited in GenBank showed differences in nucleotide sequences of repeat region in ITS1.

Conclusion

Trichobilharzia regenti is the most frequent parasite of Anatid birds in North of Iran. This corresponds to the distribution of this parasite along the flyway of migratory birds, which annually migrate from Siberia and northern countries of Caspian Sea to wintering areas in southern regions of it.     

Two New Species of the Genus Pallisentis Van Cleave, 1928 (Acanthocephala: Quadrigyridae) from the Intestine of Channa punctatus (Bloch, 1793) from the River Gomti at Lucknow,India

Background:

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.

Methods:

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.

Results:

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.

Conclusion:

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.     

PCR-RFLP Analysis of 28 SrDNA for Specification of Fasciola gigantica (Cobbold, 1855) in the Infected Lymnaea auricularia (Linnaeus, 1785) Snails from Northwestern Iran

Background

Fasciolosis in livestock is a crucial concern in the globe, mainly due to its impact on human health. The aim of this study was to determine the rate of infection with Fasciola gigantica (Cobbold, 1855) larvae in the field-collected snails of Lymnaea auricularia (Linnaeus, 1785) from northwestern Iran using a molecular approach.

Methods

A total of 6,759 pond snails were collected from 28 freshwater bodies in West Azarbaijan. PCR was performed to amplify a 618-bp fragment of the nuclear 28 SrRNA gene of Fasciola. The PCR products were digested by AvaII restriction enzyme to create restriction fragment length polymorphism (RFLP) patterns specific for the detection of F. gigantica.

Results

Of the total collected snails 496 (7.34 %) were L. auricularia, among which 4.64% (23 out of 496) were infected with a Fasciola species according to the PCR analysis. Only 2.22% (11 out of 496) of the infected snails were from the mountainous areas. The highest Fasciola infection rate recorded in the snails of a single site was 1.81% (9 out of 496 snails). Based on the RFLP pattern, F. gigantica accounted for 2.42% of the infection rates in the study sites.

Conclusion

Application of PCR-RFLP was proven to be a useful approach for valid and robust detection of the infection with F. gigantica in its intermediate host snails. These findings may therefore be applicable for establishment of the control programs against dissemination of the infection in different regions.     

An Analysis of Clinical Characteristics of Strongyloides stercoralis in 70 indigenous patients in Iran

Background

Clinical manifestations of Strongyloides stercoralis are variable from asymptomatic to hyperinfection and devastating disseminated infections. Hereby, clinical characteristics of a large series of Iranian strongyloidiasis indigenous cases are described.

Methods

The records of people referred to the Helminthological Diagnostic Laboratory of School of Public Health, Tehran University of Medical Sciences and School of Medicine, Gilan University of Medical Sciences, during 2009-2013 were reviewed. For those patients that were infected with S. stercoralis and their clinical manifestations and demographic data were available (70 cases) a checklist was prepared and data analyzed.

Results

Forty-three patients (61.4%) were male and 27 (38.6%) female. Gastrointestinal, cutaneous and pulmonary symptoms were present in 71.4%, 25.7%, and 15.7% of patients, respectively. None of them had larva currens eruption. Eosinophilia was the most prevalent reason for suspicious on S. stercoralis, but the mean was lower in elderly patients. Hyperinfection were recorded in 8 patients (11.4%), and 2 cases had disseminated infection.

Conclusion

Eosinophilia is common both in asymptomatic and symptomatic cases of strongyloidiasis, but the mean tend to lower with increase in age.     

Parasitic Infection of an Endemic Fish (Blicca bjoerkna) and an Exotic Fish (Hemiculter beucisculus) In Anzali Lagoon,Caspian Sea,Iran

Background

In Anzali Lagoon, there are some endemic and exotic fishes. The present study was conducted to compare the parasitic fauna of Blicca bjeorkna, as an endemic fish and Hemiculter leucisculus, as an introduced fish to the lagoon.

Methods

A parasitological investigation was done on 78 specimens of B. bjoerkna and 114 of H. leucisculus. The fishes were collected from August 2009 to April 2010 by the electro fishing from Anzali Lagoon.

Results

Eleven parasites species were found in 192 fish samples. The prevalence and mean intensity of parasites in each host were as follows: Parasites from B. bjorkna were Trichodina perforata (53.85%); Myxobolus musayevi (27.19%, 1±0.79); Dactylogyrus difformis (88.05%, 8±7.24) and D. sphyrna (5.18%, 0.95±0.51), Diplostomum spataceum (98.72%, 9.51±9.01), Posthodiplostomum cuticula (15.38%, 4.25±2.5), Ripidocotyle sp. (1.28%, 2±0.74); Contracaecum osculatum (17.95%, 1.64±0.79), Philometra rischta (12.8%, 1.4±0.54), and Raphidascaris acus (1.04%, 0.03±0.26). The H. leucisculus were infected with T. perforata (27.19%), D. spataceum (7.89%, 1.33±0.54), Ps. tomentosa (7.02%, 1.62±0.49) and R. acus (0.88%, 3±0.28). B. bjoerkna was presented as a new host for M. musayevi and C. osculatum, while H. leucisculus was introduced as a new host for T. perforata and Ps. tomentosa.

Conclusion

The prevalence of parasites was significantly more in native fish than that of exotic fish (P<0.05). This reduction in parasitic infection in H. leucisculus may be due to its immune system resistance, well adaptation to the new environment, host-specific limitation for endemic parasites and disability of introduced parasite to complete its life cycle in the new host as well.     

Cloning,and Molecular Characterization of Polymorphic Iranian Isolate Theileria annulata Surface Protein (Tasp)

Background

Because of the strong immunologic responses of surface protein TaSp in Theileria annulata infected host, we tried to characterize this protein in a T. annulata isolate from Iran.

Methods

The RNA prepared from T. annulata infected cells was used to produce SMART-DS-cDNA. The Double strand cDNA was then amplified with primers derived from TaSp mRNA sequences. The PCR product was cloned in pTZ57R/T vector, sequenced and registered under accession no. {"type":"entrez-nucleotide","attrs":{"text":"JQ003240","term_id":"371927237"}}JQ003240 in GenBank.

Results

The sequence analysis showed 90%–94% nucleotide sequence identity and 68%–94% amino acid homology to the corresponding sequences of TaSp gene by T. annulata, T. sp. china I, T. sp. china and T. lestoquardi and three T. annulata reported from Iran respectively. Interestingly, the sequence analysis also showed small nucleotide sequence region near the 5‘ end in which the presented TaSp protein differed very strongly from the other known TaSp sequences. For the preparation of the recombinant protein, the cDNA was cloned in pQE-32 vector, the recombinant protein was prepared and assayed by Theileria infected bovine serum.

Conclusion

The polymorphism in TaSp gene could be detected in intra- as well as inter species. The different characterized TaSp proteins had a common identic region, which may be helpful for development of broad band vaccine based on the recombinant proteins. The polymorphism in this gene, make this protein also interesting for the diagnostic purposes.