ABSTRACT
Vaccination can be a key step in controlling hydatid cyst infection in humans and livestock in endemic areas of the disease. The aim of the Present study was to determine some of the basal biochemical properties followed by prediction and screening of B-cell and MHC-binding epitopes of EgP29 protein in silico. Some of the basic physico-chemical properties along with antigenicity, allergenicity, solubility, post-translational modification (PTM) sites, subcellular localization, signal peptide, transmembrane domain, secondary and tertiary structures followed by refinement and validations were computationally determined for this protein. Also, B-cell epitopes were predicted and screened using various web servers, while MHC-binding and CTL epitopes were predicted using IEDB and NetCTL servers, respectively. The protein is a 238-residue, 27 kDa molecule, with high thermotolerance (aliphatic: 71.81) and hydrophilicity (negative GRAVY). There were several glycosylation and phosphorylation sites in the sequence, without a transmembrane domain and signal peptide. Moreover, several B-cell and MHC-binding epitopes were found in the EgP29 protein, which could be further used in multi-epitope vaccines. In conclusion, results of the present study can be a promising sign for achieving effective approaches to the preparation of a multi-epitope vaccines against echinococcosis. So, it is necessary that the effectiveness of the protein and its epitopes be evaluated in vitro and in vivo.
Subject(s)
Echinococcosis , Echinococcus granulosus , Vaccines , Animals , Humans , Echinococcosis/prevention & control , Epitopes, B-Lymphocyte , VaccinationABSTRACT
Fascioliasis is a common human-animal disease that is reported in most parts of the world. Fascioliasis is also prevalent in different provinces of Iran. Since it has done no study on the excretory/secretory and somatic immunogenic antigens profiles of adult Fasciola in Iran, the present study was performed on the Fasciola spp. collected from Mazandaran province. For this purpose, the Fasciola worm was isolated from the liver of infected sheep, then its excretory/secretory and somatic antigens were prepared from adult worms. The protein of the samples was measured by the Lowry method. Then, somatic and secretory excretions were examined by SDS-PAGE and the protein profile of the two substances was determined. To evaluate the immunogenicity, the somatic and secretory excretions antigens of Fasciola spp. were injected into white rabbits and after boosting, the blood serum of the rabbits was collected and then Western blotting was performed on them and the results were evaluated. According to the results of Western blotting, 11 somatic antigen bands with a molecular weight of 149, 122, 99, 85, 75, 65, 50, 46, 40, 37, 30 kDa and 12 protein bands of excretory/secretory antigens with molecular weights of 100, 82, 75, 70, 58, 55, 47, 40, 38, 37, 30,25 kDa were observed in adult Fasciola spp. that immunogenic, which appear to have a protective effect or can be used to prepare a diagnostic kit.
Subject(s)
Fasciola , Fascioliasis , Sheep Diseases , Animals , Rabbits , Blotting, Western/veterinary , Electrophoresis, Polyacrylamide Gel/veterinary , Fascioliasis/epidemiology , Fascioliasis/veterinary , Iran/epidemiology , Sheep , Sheep Diseases/epidemiologyABSTRACT
Leishmaniasis is one of the major health problems in most countries of the world. Millions of people around the world are at risk for the disease. Given the prevalence of this parasite in Iran and developing countries and the emergence of resistance in some cases to existing drugs, developing an effective vaccine against leishmaniasis is necessary. This research aims to design a multi-epitope vaccine derived from LACK, LeIF, GP63, and SMT antigens of Leishmania major based on the combination of bioinformatics methods. The synthesized construct with 16.86 KDa was cloned and sub-cloned in pEGFP- N1 and pLEXSY-neo2, respectively. They were then transfected in promastigotes of L. tarentolae. After confirmation of expression, immunization was carried out in 8 groups of BALB/c mice (9 mice per group) three times at two-week intervals. Cellular immune responses were assessed before and after the challenge by L. major. Furthermore, at 3rd week post-infection, the survival rate, mean lesion size, and parasite burden were assessed. All vaccinated mice demonstrated partial immunity to higher IFN-γ levels than the control groups (P<0.05). Immunized mice with cytosolic complex (G1) indicated the highest levels of IFN-ɤ and ratio of IFN ɤ/ IL-4, the lowest levels of IL-4 and IL-10 compared to control and the other groups (P≤0.05), and produced a partial Th1 immune response. Mean lesion size and parasite burden of G1 and G5 reduced significantly compared to the other and control groups post-challenge (P<0.05). The outputs of our result could be a hopeful sign in the achievement of practical approaches as multi-epitope vaccines against Leishmania major.
Subject(s)
Leishmania major , Leishmaniasis , Animals , Mice , Epitopes , Interleukin-4 , Mice, Inbred BALB C , Th1 Cells , Vaccines, SubunitABSTRACT
Visceral leishmaniasis is a neglected disease caused by Leishmania infantum and transmitted via female sand flies. Canine visceral leishmaniasis diagnosis should be performed as soon as possible, even on the basis of only a few or even a single clinical sign, to enhance the prediction of disease and to avoid both dog and human transmission and unnecessary euthanasia of apparently positive dogs. In the present work, we examined whether PQ10 recombinant protein could be suitable for immunological detection of Leishmania infantum infection. The coding sequence of PQ10 recombinant protein was sub-cloned in pET28 expression vector and was commercially synthesized by GENERAY Biotechnology, China. In the following process, sequencing with proper primers was done and the expression, optimization of expression and protein purification were performed. The efficacy of PQ10 for serodiagnosis was evaluated with 100 serum samples collected from dogs living in the visceral leishmaniasis endemic areas of Iran. Samples (n=20) of the dogs with other infectious disease were also be collected. The synthesized colones verified by the sequencing with proper primers. In the following process, expression, optimization of expression and protein purification performed and the purified recombinant protein confirmed by western blot. The ELISA was performed with PQ10 recombinant protein. The sensitivity of ELISA that was evaluated with sera from naturally infected dogs was 94%. The specificity value of the ELISA determined with sera from healthy dogs and from dogs with other infectious diseases was 86%. The positive predictive value (PPV) and negative predictive value (NPV) determined 87.03% and 93.47% respectively. Our findings indicated to the potential use of this recombinant protein in the diagnosis of canine visceral leishmaniasis.
Subject(s)
Diagnostic Tests, Routine/veterinary , Disease Reservoirs/veterinary , Dog Diseases/diagnosis , Enzyme-Linked Immunosorbent Assay/veterinary , Leishmania infantum/isolation & purification , Leishmaniasis, Visceral/veterinary , Protozoan Proteins/analysis , Animals , Diagnostic Tests, Routine/methods , Disease Reservoirs/parasitology , Dog Diseases/parasitology , Dogs , Enzyme-Linked Immunosorbent Assay/methods , Leishmaniasis, Visceral/diagnosis , Leishmaniasis, Visceral/parasitology , Recombinant Proteins/analysisABSTRACT
Neospora caninum, a protozoan parasite, causes abortions in cattle, as well as neurological disorders and reproductive problems in dogs. This study aimed to investigate the serological and the molecular prevalence of N.caninum among foxes and dogs using indirect fluorescent antibody test (IFAT) and nested-polymerase chain reaction (PCR). For this purpose, 288 and 95 both fecal and serum samples of dogs and foxes were collected, respectively, from around industrial and traditional dairy flocks in different parts of Sanandaj, Kurdistan Province, Iran, from 2013 to 2015. The sera were examined using IFAT, and fecal samples were microscopically assessed for detecting Neospora oocyst and by nested-PCR. The findings revealed that N.caninum seroprevalence were 4.86% and 4.21% in dogs and foxes, respectively. In addition, no Neospora oocysts were found microscopically and by PCR. Since this study is the first serological and molecular investigation of N.caninum among both dogs and foxes in Sanandaj, the findings of indicated that stray dogs is a main source of N.caninum infection in dairy farms in Sanandaj, Iran.
Subject(s)
Coccidiosis/veterinary , Foxes , Neospora/immunology , Animals , Coccidiosis/epidemiology , Dog Diseases/epidemiology , Dogs , Female , Fluorescent Antibody Technique, Indirect/veterinary , Iran/epidemiology , Male , Polymerase Chain Reaction/veterinary , Prevalence , Seroepidemiologic StudiesABSTRACT
Hepatozoonosis is a protozoal disease caused by various species of Hepatozoon. This parasite is transmitted from tick; the main vector of Hepatozoon canis is usually the brown dog tick (Rhipicephalus sanguineus). However, several species of ticks are disposed as the alternative vectors. Dogs are usually infected by eating the tick or a part of the tick organ infected by the mature oocysts containing infectious sporozoite. In the current study, a total of 145 blood samples were collected from the cephalic vein of pet, stray, and shelter dogs in Tehran. To conduct this study, first thin blood smears were prepared from all the samples and stained with the Giemsa method. Then, after extraction of DNA from the blood samples, in order to trace Hepatozoon canis, the 18S rRNA gene segment of the parasite was amplified using polymerase chain reaction (PCR). To confirm the PCR-positive results, five randomly selected PCR-positive samples were sequenced. According to the results, through direct observation of microscopic slides, no infection of H. canis parasite was observed, but according to the PCR results, 32 out of the 145 blood samples were found to be infected by H. canis. In this study, infection to H. canis in older dogs was higher than in young dogs, and more male dogs were found to be infected by the parasite compared to female dogs; but no significant difference was observed in this regard (P > 0.05). Moreover, stray dogs showed a significantly higher rate of infection, compared to the pet and shelter ones (P < 0.05).
Subject(s)
Coccidiosis/veterinary , Dog Diseases/epidemiology , Eucoccidiida/isolation & purification , Age Factors , Animals , Coccidiosis/epidemiology , Coccidiosis/parasitology , Dog Diseases/parasitology , Dogs , Female , Genes, rRNA , Iran/epidemiology , Male , Polymerase Chain Reaction/veterinary , Prevalence , RNA, Protozoan/analysis , RNA, Ribosomal, 18S/analysis , Sex FactorsABSTRACT
In this experimental study, first the killing effect of silver nanoparticles alone or in combination with 3mA of Half-Wave Rectified Sine current was assessed in promastigote culture for 10 minutes. The survival rate of infected promastigotes was evaluated by Flow cytometery. In the second step, BALB/c mice were infected experimentally with L. major, followed by silver nanoparticles injected inter-lesion and simultaneously 3 mA a Half-Wave Rectified Sine current induction was applied directly into the wound. Finally, the lesion size and the mice body weight changes were measured during 5 weeks. Results indicated that simultaneous use of nanoparticle and electricity increased the mortality of promastigotes significantly. However, when 3 mA of HWRS and 160 µm/ml nanosilver were used alone in medium culture only 73.4% and 32% of promastigotes were killed respectively but the combined use destroys the promastigotes completely. The diameter of the lesions after six weeks in the control group; group treated with meglumine antimoniate and the group treated with HWRS increased to 6.01, 0.02 and 0.52 mm but in group treated with HWRS plus Nanosilver was reduced to -0.14 mm. The results showed that, when silver nanoparticles with HWRS current electricity were used in mice, the skin lesions were reduced in size but like Glucantime, complete healing was not achieved.
ABSTRACT
Vaccination would be the most important strategy for the prevention and elimination of leishmaniasis. The aim of the present study was to compare the immune responses induced following DNA vaccination with LACK (Leishmania analogue of the receptor kinase C), TSA (Thiol-specific-antioxidant) genes alone or LACK-TSA fusion against cutaneous leishmaniasis (CL). Cellular and humoral immune responses were evaluated before and after challenge with Leishmania major (L. major). In addition, the mean lesion size was also measured from 3th week post-infection. All immunized mice showed a partial immunity characterized by higher interferon (IFN)-γ and Immunoglobulin G (IgG2a) levels compared to control groups (p<0.05). IFN-γ/ Interleukin (IL)-4 and IgG2a/IgG1 ratios demonstrated the highest IFN-γ and IgG2a levels in the group receiving LACK-TSA fusion. Mean lesion sizes reduced significantly in all immunized mice compared with control groups at 7th week post-infection (p<0.05). In addition, there was a significant reduction in mean lesion size of LACK-TSA and TSA groups than LACK group after challenge (p<0.05). In the present study, DNA immunization promoted Th1 immune response and confirmed the previous observations on immunogenicity of LACK and TSA antigens against CL. Furthermore, this study demonstrated that a bivalent vaccine can induce stronger immune responses and protection against infectious challenge with L. major.
Subject(s)
Antigens, Protozoan/immunology , Leishmaniasis Vaccines/immunology , Leishmaniasis, Cutaneous/immunology , Peroxiredoxins/immunology , Protozoan Proteins/immunology , Vaccines, DNA/immunology , Animals , Female , Leishmania major , Mice, Inbred BALB C , Plasmids , Recombinant Fusion Proteins/immunologyABSTRACT
Hepatozoon species are protozoan parasites that infect some animals such as birds, reptiles, amphibians, and carnivores. Previous studies performed on canine hepatozoonosis in Iran have never used molecular techniques for diagnosis of this disease. The main objective of the present study was to detect Hepatozoon canis in the blood of dogs using polymerase chain reaction (PCR) method and sequencing. A total of 104 blood samples were collected from dogs of Meshginshahr County (Ardabil Province), and DNA was extracted from blood samples by dint of DNG-plus Extraction Kit. Then, 18S rRNA gene was amplified by using the conventional PCR methods. PCR products yielded an amplicon of the approximate length of 897 bp for all the positive samples. Twenty-four out of the 104 (23.07%) samples were found to be positive for H. canis. This rate of infection is relatively high among dogs in Ardabil Province. Sequence analysis confirmed the molecular identity of 99% of the samples by comparison with GenBank profiles. This is the first report of molecular detection of H. canis from Iran.
Subject(s)
Coccidiosis/veterinary , Dog Diseases/epidemiology , Eucoccidiida/isolation & purification , Animals , Coccidiosis/blood , Coccidiosis/epidemiology , Dog Diseases/blood , Dogs , Iran/epidemiology , Prevalence , RNA, Protozoan/analysis , RNA, Protozoan/blood , RNA, Ribosomal, 18S/analysis , RNA, Ribosomal, 18S/blood , Sequence Analysis, RNA/veterinaryABSTRACT
Cutaneous leishmaniasis is one of the main public health problems in Afghanistan, particularly in Herat. To identify Leishmania spp., molecular techniques were applied to samples from 64 cutaneous leishmaniasis patients referred to Herat regional hospital during 2013. Polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) analysis of the ribosomal RNA gene internal transcribed spacer-1 (ITS1) was used. Most of the patients demonstrated dry type single lesions on the head. The results of direct microscopy detection using Giemsastained skin scrapings were compared with that of ITS PCR-RFLP for the diagnosis of cutaneous leishmaniasis. Light microscopy examination showed 37/64 positive cases (58%). PCR revealed 50 positive cases (78%), from which ITS PCR-RFLP identified 48 cases (96%) as L. tropica and 2 cases (4%) as L. major. Cutaneous leishmaniasis in Herat appears to be endemic and of the clinically dry type, caused mainly by L. tropica and occasionally by L. major.
Subject(s)
Leishmania/classification , Leishmaniasis, Cutaneous/parasitology , Adolescent , Adult , Afghanistan , Aged , Aged, 80 and over , Child , Child, Preschool , Female , Humans , Infant , Leishmania/isolation & purification , Male , Middle Aged , Polymerase Chain Reaction , Polymorphism, Restriction Fragment LengthABSTRACT
Previous studies have shown that Leishmania elongation initiation factor (LeIF) antigen causes a partial immunity against leishmaniasis. The antigen develops type I immunity by overexpression of inflammatory cytokines such as interleukin-12 (IL-12), IFN-γ and TNF-α. Therefore, We evaluated immune responses induced by the LeIF gene against Leishmania major infection. Immunization with LeIF gene alone or with IL-12 induced Th1 response and produced higher IFN-γ and lower IL-4 levels by splenocytes than control groups (P < 0·05) and also ratios of IFN-γ/IL-4 were 11·68 to 18·53 times more in immunized groups than control groups after challenge. In addition, analysis of humoral immune response revealed that immunized mice had more IgG2a levels than both control groups (P < 0·05). On the other hand, lesion size was less for immunized animals than control groups from 4th week after challenge (P < 0·05). The percentage reduction in lesion size was 29·30% for LeIF and 51·98% for LeIF + IL-12 than PBS at 12th week post-infection. Spleen parasite burden decreased in all immunized groups in comparison with control groups (P < 0·05). The results indicated that LeIF gene induced partial immunity against L. major infection in BALB/c mice. However, LeIF plus IL-12 group showed more potent immunity with smaller lesions than other groups.
Subject(s)
Leishmania major/immunology , Leishmaniasis, Cutaneous/prevention & control , Vaccination , Animals , Antibodies, Protozoan/blood , Cytokines/immunology , Female , HEK293 Cells , Humans , Immunity, Humoral , Interleukin-12/genetics , Interleukin-4/biosynthesis , Leishmania major/genetics , Leishmaniasis, Cutaneous/immunology , Leishmaniasis, Cutaneous/parasitology , Mice , Mice, Inbred BALB C , Peptide Elongation Factors/genetics , Protozoan Proteins/genetics , Spleen/immunology , Spleen/parasitology , Transfection , Tumor Necrosis Factor-alpha/biosynthesis , Vaccines, DNA/administration & dosageABSTRACT
OBJECTIVE: Leishmaniasis is usually treated with chemotherapy; however, toxicity, resistance and high-cost limit use of the chemical drugs. Leishmania eukaryotic initiation factor (LeIF) protein acts the same as interleukin (IL)-12 and reduces the secretion of IL-4 in lymph node cells of mice infected with Leishmania major. The aim of this study was cloning of the gene encoding LeIF antigen into eukaryotic expression plasmid pEGFP-N1. METHODS: DNA was extracted from Iranian strain of the L major (MRHO/IR/75/ER) promastigotes. The full-length sequence of LeIF was amplified with Pfu DNA polymerase using a specific primer. The amplified LeIF was cloned into a pJET1.2/blunt vector. Then this fragment was digested with HindIII and EcoRI and was subcloned into the pEGFP-N1 vector. Confirmation of the cloning was done by colony polymerase chain reaction (PCR). RESULTS: Leishmania eukaryotic initiation factor gene was successfully cloned and subcloned into pJET1.2 and pEGFP-N1 plasmids, respectively. The results of colony PCR, restriction analysis and sequencing confirmed them. CONCLUSIONS: We cloned LeIF gene which could be expressed in eukaryotic cells in vivo and could be used as a vaccine candidate against leishmaniasis in future studies.
ABSTRACT
Toxoplasmosis is one of the most common parasitic diseases worldwide. Although estimated that one third of the world's population are infected with Toxoplasma gondii, but the most common form of the disease is latent (asymptomatic). On the other hand, recent findings indicated that latent toxoplasmosis is not only unsafe for human, but also may play various roles in the etiology of different mental disorders. This paper reviews new findings about importance of latent toxoplasmosis (except in immunocompromised patients) in alterations of behavioral parameters and also its role in the etiology of schizophrenia and depressive disorders, obsessive-compulsive disorder, Alzheimer's diseases and Parkinson's disease, epilepsy, headache and or migraine, mental retardation and intelligence quotients, suicide attempt, risk of traffic accidents, sex ratio and some possible mechanisms of T. gondii that could contribute in the etiology of these alterations.
ABSTRACT
BACKGROUND: Echinococcosis or hydatidosis is a chronic, zoonotic worldwide infection that occurs by the larval stages of taeniid cestodes of the genus Echinococcus. Iran is known as endemic region for this infection in the world. Vaccination has been considered as a good prevention method for this disease. Recombinant vaccines containing EG95 protein, against E. granulosus, has shown a high degree of protection against E. granulosus infection. In this study EG95 gene was extracted from Iranian isolates of E. granulosus and then cloned and expressed in expression vector. METHODS: Protoscoleces were collected from sheep hydatid cysts. Then DNA and RNA were extracted from protoscoleces, and amplified by PCR and RT-PCR with specific primer. Afterward the purified RT-PCR products were successfully ligated into pTZ57R/T plasmid vector. The pcDNA3 plasmid was used as expression vector and Eg95 fragment sub cloned into this plasmid. The pcEG95 plasmid was digested by restriction enzymes to confirm cloning of this gene in pcDNA3 plasmid. In last step, the subcloned gene was expressed in CHO as eukaryotic cell. RESULTS: EG95 fragment successfully was subcloned in pcDNA3 and EG95 protein was expressed by eukaryotic cell. The recombinant EG95 protein was confirmed by SDS-PAGE and Western blot. CONCLUSION: Recombinant plasmid of pcEG95 was constructed successfully and express of recombinant EG95 protein was confirmed.
ABSTRACT
Toxoplasmosis is one of the classical conditions known to have an adverse effect on female reproductive functions, but a few investigations into male reproductive parameters have been performed. This work was carried out to study the effects of Toxoplasma gondii on reproductive function in male rats. Male rats were infected with the RH strain of T. gondii tachyzoites, and following every 10 days from 10 to 70 postinfection (PI), the percentage of body weight to testis weight ratio as well as epididymal sperm parameters (number, motility, viability, and morphology rates), serum testosterone (ST), intratesticular testosterone (ITT), serum lactate dehydrogenase (SLDH), intratesticular lactate dehydrogenase and fructose in seminal vesicles and coagulating glands were measured. The results of the study showed sperm motility, viability and concentration rates were significantly decreased temporary after infection up to 70 days. Sperm abnormality was also increased during these days. In addition, temporary alteration in ST, ITT, SLDH, intratesticular LDH and fructose in seminal vesicle and coagulating gland was observed PI. These findings suggest that toxoplasmosis can cause impermanent impairment on the reproductive parameters of male rats.
Subject(s)
Reproduction , Toxoplasma/isolation & purification , Toxoplasmosis/physiopathology , Animals , Male , RatsABSTRACT
Toxoplasma gondii, the pathogen of toxoplasmosis, can infect most mammals and birds. The high incidence and severe or lethal damages of toxoplasmosis clearly indicate the need for the development of a more effective vaccine. We constructed a DNA cocktail, containing plasmids encoding the full-length SAG1 and ROP2 genes of T. gondii and evaluated its immune response and protective efficacy in comparison with single-gene vaccines and control groups. We immunized BALB/c mice intramuscularly three times. DNA cocktail elicited IgG and IFN-γ, TNF-α and IL-2 greater than single-gene plasmids and increased survival time against a lethal challenge with the highly virulent T. gondii RH strain. The current study shows that pc-SAG1+ pc-ROP2 as a cocktail DNA vaccine produces higher Th1 immune response than single-gene plasmids and cocktail DNA is effective to prime an enhanced and balanced specific immunity.
Subject(s)
Antigens, Protozoan/immunology , Membrane Proteins/immunology , Protozoan Proteins/immunology , Toxoplasma/immunology , Toxoplasmosis/prevention & control , Vaccines, DNA/immunology , Animals , Antibodies, Protozoan/blood , Antigens, Protozoan/genetics , Female , Immunization, Secondary/methods , Immunoglobulin G/blood , Injections, Intramuscular , Interferon-gamma/metabolism , Interleukin-2/metabolism , Membrane Proteins/genetics , Mice , Mice, Inbred BALB C , Plasmids , Protozoan Proteins/genetics , Survival Analysis , Toxoplasma/genetics , Toxoplasmosis/immunology , Toxoplasmosis/mortality , Tumor Necrosis Factor-alpha/metabolism , Vaccination/methods , Vaccines, DNA/administration & dosage , Vaccines, DNA/geneticsABSTRACT
BACKGROUND: Hyalomma anatolicum is the well-known hard tick, which is one of the most important livestock and human pathogens vector, wide range in host and distributed in all over the Hyalomma geographic fauna as well as in Iran. Taxonomy of the Hyalomma ssp. is debatable whereas their identification is a problematic work. The reasons for this claim is time consuming Delpy's researches in Iran also Schulze School, Feldman-Muhsam and the Russian tick workers. We would like to understand morphometric variation in the field collected H. anatolicum in Iran also validating some morphologic quantitative and qualitative characters. METHODS: A total 247 field-collected tick specimens from different geographical regions in west of Iran includes Khuzestan and Lorestan Provinces were studied. The morphologic characters of the ticks were measured by the calibrated stereomicroscope armed scaled lens. The measurements were analyzed using SPSS for windows, version 16 on an IBM PC, so varied shapes of species in different geographic regions were drawn by the aid of a drawing tube connected to a light stereomicroscope. RESULTS: One way ANOVA test revealed significant differences among the quantitative parameters in five zones (P < 0.001) also each zone to other zone by Post Hoc Tests e.g. LSD. No significant differences in the lateral grooves length/conscutum length ratio parameter were found. CONCLUSION: Morphometric variation in Hyalomma spp is poorly studied. The variation in range and quantity of the morphometric parameters of H.anatolicum underlies that the correct recognition and key construction for Hyalomma species dependes on a complement morphometric study on the other species.
ABSTRACT
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.
ABSTRACT
Human infection especially with helminth parasites is an emerging health issue, as the human environment is increasingly shared with infected animals, either pets or wild life. In this survey, the intestinal content of 83 stray dogs, 22 red foxes and 10 golden Jackals collected from the West Azarbaijan, Kordestan and Kermanshah provinces in the west of Iran, were studied for the presence of helminth parasites. The percentage of different species recovered from these animals is listed as follows: From stray dogs: Toxocara canis (6.02%), Toxascaris leonina (32.53%), Ancylostoma caninum (3.61%), Oxynema sp. (1.35%), Rictularia affinis (12.05%), Taenia hydatigena (53.01%), Taenia ovis (7.23%), Taenia multiceps (4.82%), Echinococcus granulosus (13.25%), Dipylidium caninum (38.55%), Mesocestoides lineatus (26.50%) and Macracanthorhynchus hirudinaceus (4.82%). From red foxes: T. canis (4.54%), T. leonina (31.82%), A. caninum (4.54%), Uncinaria stenocephala (13.64%), Oxynema sp. (9.09%), R. affinis (54.54%), Strongyloides sp. (4.54%), Physaloptera sp. (4.54%), T. hydatigena (9.09%), E. granulosus (4.54%), D. caninum (9.09%), M. lineatus (81.82%), Joyeuxiella pasqalei (27.27%), Diplopylidium nolleri (4.54%), M. hirudinaceus (22.72%) and Macracanthorhynchus sp. (9.09%). From golden jackals: T. canis (10%), T. leonina (30%), R. affinis (50%), T. hydatigena (10%), D. caninum (20%), M. lineatus (70%), J. pasqalei (30%.), Alaria canis (10%), M. hirudinaceus (30%) and Macracanthomynchus sp. (10%).
Subject(s)
Dog Diseases/epidemiology , Foxes/parasitology , Helminthiasis, Animal/epidemiology , Intestinal Diseases, Parasitic/veterinary , Jackals/parasitology , Animals , Dog Diseases/parasitology , Dogs , Helminthiasis, Animal/parasitology , Intestinal Diseases, Parasitic/epidemiology , Intestinal Diseases, Parasitic/parasitology , Iran/epidemiologyABSTRACT
In the present study, Echinoccocus granulosus isolates collected from human, sheep and camel samples in Iran were characterized based on rostellar hook morphology of protoscoleces as well as PCR-RFLP. Morphological study on human and animal isolates showed the presence of two distinct strains of the parasite, one in sheep and the other one in camels. In this regard, rostellar hook of sheep isolates were significantly different from those of camel origin, meanwhile human isolates were found to be similar to those isolated from sheep. Molecular analysis of the ITS1 region of rDNA derived from human, sheep and camel isolates were in agreement with the morphological findings. Based on the PCR-RFLP method, the sheep and human isolates appeared to pertain to the same genotype and the camel isolates were appeared to pertain to a different genotype.