Inhibition of murine systemic leishmaniasis by acetyl salicylic acid via nitric oxide immunomodulation

The purpose of this study was to evaluate antileishmanial effects of ASA via NO pathway in Leishmania major infected Balb/c mice. Moreover, toxicity and pathological consequences of ASA administration were investigated. Balb/c mice were infected with L. major and ASA was inoculated orally after lesion appearance for its ability to modulate NO and to modify Leishmania infection in host, in order to evaluate the effects of NO production on size and lesion macroscopy, delay of lesion formation and proliferation of amastigotes inside macrophages. Liver, spleen, and lymph nodes were also studied as target organs to detect amastigotes. In addition, plasma was investigated for NO induction using Griess microassay. ASA increased NO production in plasma of both na‹ve and Leishmania test groups at the ultimate of the experimental period. A decline was observed in proliferation of amastigotes inside macrophages of test group when compared with control one. ASA reduced lesion size, inhibited Leishmania visceralisation in spleen, lymph node, and decreased hepato/splenomegaly in ASA treated animals. Some antileishmanial effects of ASA by NO-modulation were indicated during systemic leishmaniasis in mice. Despite slight effects on lesion size, ASA decreased parasite visceralization in target organs and declined their proliferation inside macrophages. Therefore, ASA may be indicated to inhibit systemic leishmaniasis via NO pathway in mice model

[Isolation and molecular typing of wolbachia pipientis from two species of phlebotomus caucasicus and phlebotomus mongolensis sandflies]

In Iran, zoonotic cutaneous leishmaniasis [ZCL] is an endemic disease in many foci in the northeastern, southern, and central parts of the country. This disease goes through the geographical distribution along with dispersion in their reservoirs [gerbils] and their vectors [sandflies]. Therefore, controlling the vectors or reservoirs has a significant role in prevention of Leishmania parasites which is transmitted by sandflies. Nowadays, because of vectors implications, the routine methods of controlling and spraying has no more useful effects on vectors and reservoirs. Consequently, in recent years maternally inherited intracellular Rickettsia like bacteria [Wolbachia] has been fascinated by many researchers. The aim of the present study was to improve our knowledge about detection of two species of Paraphlebotomus sandflies infected with W. pipientis which yet has not been reported in Iran and the world. The new surveys have been conducted in the case of Wolbachis detection in two mentioned ZCL vectors. In Turkemen Sahara within the ZCL focus, two species of Phlebotomus cauccasicus and Phlebotomus mongolensis sandflies has been frequently collected from eighteen villages. Sticky papers and CDC traps were used to sampling sandflies in rural areas. In the laboratory, sandflies were identified to species by dissecting and mounting genitalia of each sandfly. DNA from sandflies [Thorax and abdomen] was extracted, the wsp gene confirmed for the presence of Wolbachia using wsp general primers [81F/691R]. After sequencing, the data were analyzed by molecular software. We examined a total of 136 individuals [91 male and 45 males] from Phlebotomus caucasicus and Phlebotomus mongolensis species; 10 out of 44 positive [32.35%] samples had enough DNA to sequencing. Wobachia infections have been found and verified for the first time in each of two Phlebotomus caucasicus and Phlebotomus mongolensis species in Iran and the world. In this procedure, 3 haplotypes [2 common Haplotypes and 1 unique Haplotype] of 2 species of Paraphlebotomus subgenus has been recognized in 10 sand flies of Iran. Paraphlebotomus sandflies are the secondary vectors of ZCL after Phlebotomus which play a decisive role in maintaining disease of their reservoirs. Wolbachia provide a starting point for inducing changes in host sex or sexuality. By manipulating Wolbachia as a transgene, it is hoped that these bacteria may be used as controlling system for decreasing vector-borne-disease

Seroepidemiology of Herpes Simplex Virus Type 1 and 2 in Northern Iran

Herpes simplex virus [HSV] type 1 and 2 are common infectious agents worldwide. Data on prevalence of HSV-1 and HSV-2 infection are limited in Asia, especially in Iran. The aim of this study was to determine the seroprevalence of HSV type 1 and 2 based on age, gender, marital status, education, living area, job, symptoms and history of disease variables. The study population included 800 randomly selected persons from laboratories in Gilan Province, Iran, from 2010 to 2011. Demographic data gathered by a well-designed questionnaire and for serological studies, blood samples were collected and centrifuged. ELISA HSV-1, 2 and HSV-2 specific ELISA kits were used to determine IgG type specific antibodies in sera samples. Person's chi-square test was applied to compare HSV-1 and HSV-2 seropositivities. HSV-1 and HSV-2 IgG antibodies wrere positive in 467 [58.4%] and 28 [3.5%] subjects, respectively. There was significant correlation between age, marital status, job, symptoms, history of disease and HSV seroprevalence [P<0.05]. Our findings were in agreement with prior studies in which HSV-1 infections was more prevalent than HSV-2 and seropositivity increased with age

Aflatoxin M1 in pasteurized milk in Babol city, Mazandaran province, Iran

Aflatoxin M1 [AFM1] is the metabolite of aflatoxin B1 [AFB1] and is found in milk when lactating animals are fed with contaminated feedstuff. The presence of AFM1 in milk, pose a major risk for humans especially kids as it can have immunosuppressive, mutagenic, teratogenic and carcinogenic effects. The present study is aimed to investigate the occurrence of AFM1 in subsidized pasteurized milk in Babol, Mazandaran Province, Iran. Some 72 pasteurized milk packages were collected from supermarkets in various districts of city during January to March 2006. Milk samples were centrifuged and amounts of 100 micro l of skimmed milk were tested for AFM1 contamination by competitive ELISA. All the samples [100%] exhibited contamination with AFM1. The contamination levels means in January, February, and March were 227.85, 229.64, and 233.1ng/l, respectively. The amount of AFM1 in all the samples were above 50ng/l, the threshold set by the European community regulations. Monitoring of AFM1 level should be part of quality control procedures in dairy factories, particularly the ones providing infant's milk. Production of safer and healthier milk and other dairy products with minimum AFM1 level can be achieved by adopting prophylactic measures including control of humidity and water content of feedstuff, which favors mould production

Molecular diagnosis of trichomoniasis in negative samples examined by direct smear and culture

Trichomoniasis is an extremely common sexually transmitted infection [STI] worldwide and is associated with important public health problems, including amplification of HIV transmission. This disease is in forms of symptomatic and asymptomatic in women and may depend on host as well as parasite variables. Most of the studies reported from females are based on examination of vaginal secretions and urine samples by direct smear and culture in modified Diamond's media. The aim of this study was checking the samples, which were negatively by direct smear and culture, with PCR technique. The urine samples and vaginal discharge of patients attending Gynecology Clinics of Mazandaran Province, Iran with different symptoms rechecked for Trichomonas vaginalis by PCR technique using primers targeting a conserved region of the beta-tubulin genes of the parasite. Data were analyzed by Epi Info software program. Out of 161 negative samples by direct smear and culture, seven samples [4.3%] were positive by PCR technique. Diagnosis of trichomoniasis by PCR is a sensitive and specific method that could play important role to help the physicians for properly treatment and control of infection

[Seroparasitological evaluation of Plasmodium vivax malaria and stability of the anti-plasmodial antibodies in Parsabad, Ardabil province]

Malaria is one of the most important parasitic diseases in the world and a major health problem in some areas of Iran. In addition to endemic areas in the south and south-eastern part of Iran, a new threat of Plasmodium vivax malaria importation emerged from the Parsabad district, which is located in Ardabil province in the north western part of the country. Malaria in this area may have originated from Azerbaijan, Armenia or southern part of Iran. This study has been carried out to clarify seroparasitological results from Indirect Fluorescence Assay [IFA], stability of antiplasmodial antibodies and its comparison with those of confirmed direct microscopy in Parsabad district during 2003-2005. This seroparasitological study has been carried out on 250 samples from malaria infected patients which was previously confirmed by microscopy and treated with routine antimalarial agents, and 250 samples of healthy control with no history of malaria in Parsabad during two years [2003-2005]. Sera of collected blood samples were assessed for the presence of anti-plasmodial antibodies using IFA assay. Statistical analysis was applied by using ANOVA and Students t-tests with Graph Pad Prism. The results of this study indicated that all blood smears of test group were detected as positive by observation of P. vivax by direct microscopy and no positive smears were found among control group. Moreover, no mixed-infection was observed among collected samples. In addition, serological results revealed that 47 cases [19%] from test group and 4 cases [1.6%] from control group had antibodies against P. vivax malaria [P<0.001]. The results of this study demonstrated that the rate of antiplasmodial antibodies is not stable in malaria infected patients which was previously confirmed by microscopy and can not be used for epidemiological evaluation for malaria in this area. Therefore, more investigation is needed for evaluation and detection of the malaria

Trichomoniasis in asymptomatic patients

Trichomoniasis is an extremely common infection worldwide and is associated with important public health problems, including amplification of HIV transmission. This disease is in forms of symptomatic and asymptomatic in women and may depend on host as well as parasite variables. Thus the aim of this study was to evaluate the patients attending gynecology clinic with different symptoms and checked them for Trichomonas vaginalis infection. The vaginal secretion and urine samples of the patients attending gynecology clinic, Imam Reza Hospital, Amol City, were checked by direct smear and cultured in TYI-S-33 culture media. Out of 853 samples collected, 8[0.9%] were positive for T. vaginalis and the number of asymptomatic patients for trichomoniasis were 1.3 times more than infected samples belong to symptomatic ones. Per speculum examination revealed that 75% of T. vaginalis positive subjects had normal appearance of vagina and cervix. According to the epidemiological aspects, these asymptomatic patients are very important as healthy carriers, and T. vaginalis infections are commonly associated with other STDs and are a marker of high-risk sexual behavior. Thus laboratories could play important role for diagnosis of infection and help the physicians for proper treatment

Characterization of specific IgE antibody related to antigen 5 of echinococcus granulosus

Anaphylactic reactions, such as urticaria, edema, respiratory symptoms, and anaphylactic shock often complicate the course of Cystic Echinococcosis [CE]. To investigate the role of the IgE immunoreactive antigen 5 [Ag 5] in the sero-positive patients with CE, we determined N-terminal of 57 kDa subunit of Ag5 responsible for IgE and C-terminal of this active antigen related to induction of IgG specifically. Immunoblotting analysis showed that specific IgE to 57-kDa subunit related to inter-chain disulphide band of two 22 kDa and 38-kDa component of Ag5 and conformational epitope on this subunits. In addition, since the 57 kDa component arise from the removal of the C-terminal portion of 22 kDa subunit of Ag5, thus IgE specifically recognized N-terminal of 22 kDa subunit which remain bounds to the other component, whereas IgG reacted with C-terminal of 38 kDa component of Ag5. Recognition of the specific binding site on the 57 kDa subunit of Ag5 could leads to understanding the mechanism regulating IgE/IgG production in some immune circumstances that IgE tends to some dominate, whereas in other IgG predominates

Distribution of soft ticks and their natural infection with Borrelia in a focus of relapsing fever in Iran

Tick-borne diseases such as relapsing fever and Crimean-Congo Hemorrhagic Fever [CCHF] are of public health importance in Iran. There are 471 reported cases of relapsing fever in 2003, according to the Ministry of Health of Iran. The number of cases has been increased in recent years. Its distribution is more or less prevalent in different parts of Iran. The aim of this study was to find out the fauna and natural infection of soft ticks with Borrelia in Qazvin Province, during their seasonal activity. The province covers 15821 km[2] between 48-45 to 50-50 east of Greenwich Meridian of longitude and 35-37 to 36-45 north latitude of the equator. For this purpose a field study was carried out in the region. A total of 54 villages from 19 districts were selected randomly and ticks were collected from their habitats according to the standard method. A total of 3197 Argasidae ticks were collected from human dwellings, poultry and animal shelters. They belonged to Argas and Ornithodoros genera which 36.8% were Argas persicus, 4% A. reflexus, 6.4% O. canestrini, 45.5% O. lahorensis and 7.3% O. tholozani. It should be noted that 12 ticks of O. erraticus were collected from 12 rodents borrows. We found that 8.82% of O. tholozani ticks were infected with Borrelia persica and half of the O. erraticus were infected with Borrelia microti. All the people who are involved with veterinary activities should be aware of disease transmission by the ticks. In the endemic area of the disease tick control is recommended

Nuclear elongation factor-I alpha gene a molecular marker for Iranian sandfly identification

Elongation factor-1 alpha, a conserved nuclear protein coding gene was used to identify Iranian sandfly species. The phlebotomine sand flies are the vectors of the parasitic protozoan Leishmania, the causative agents of leishmaniasis, in Iran. Seven sets of primers were tried. PCR amplification of elongation factor-l alpha was successfully achieved for all 14 species of Iranian sandflies that we caught, but different primers had to be used. The aligned DNA sequences of 454 bp [without primers] of the gene had the most similarity to a coding region of the elongation factor-l alpha genes of D. melanogaster, as identified by a BLAST search of GenBank. Each Iranian species, except Phlebotomus caucasicus and P. mongolensis, had a unique combination of nucleotides, i.e. each had a diagnostic sequence. There were no diagnostic sequences for different geographical populations of the species in Iran. We found only a single copy of Ef-l alpha gene in most individual sandflies. However EF-l alpha gene was successfully amplified by PCR but, unfortunately, phylogenetic analysis showed that it might be multicopy in sandflies and so the markers could not be trusted. More highly polymorphic nuclear loci, like microsatellites, might be needed to distinguish morphologically indistinguishable females of the subgenus Paraphlebotomus, e.g. P. caucasiscus from P. mongolensis, in order to resolve their roles as vectors of Leishmania species in gerbils

study on leishmania infection rate among phlebotomus spp. collected from Abardejh district, Iran

Cutaneous leishmaniasis is a prevalent tropical parasitic disease in the Old World. The causative agents are Leishmanial parasites, which cause various forms of cutaneous leishmaniasis. The infection is commonly limited in immuno-competent individuals, but it can progress to a chronic and ulcerative disease in immunocompromised patients. The reservoirs are dogs and rodents and the vectors are different species of sandflies. In the present study, we investigated the prevalence rate of Leishmania infection among Phlebotomus mosquitoes collected from Abardejh district, Iran. Abardejh is located next to Varamin city in southeast of Tehran having a tropical ecosystem at its eastern border. Tamarisk trees and rodents have provided a suitable condition for sandfly activity. The sandflies were collected by funnel trap from rodent burrows and transferred to the Department of Parasitology, Pasteur Institute of Iran. The sampling was carried out during spring and summer [2002] with ten-day intervals. The collected sandflies were identified using discriminative morphologic features before parasitological culture on NNN medium. Analyses of the data revealed a high prevalence rate of infection among the sandflies in this region [P<0.01]. The maximum activities of Phlebotomus were in the months of June and July. Three species of sanflies were found in rodent burrow: P. papatasi, P. sergenti, and P. caucasicus. The results of blood-fed Phlebotomus culture showed that 22.07% of blood-fed females of P. papatasi and 8% of blood-fed females of P. sergenti were infected with leptomonads [P<0.05]. This could be an important issue because human and agricultural environments are located closely to this district. Therefore, use of insecticides and environmental sanitation seems to be required to prevent the transmission of infection from sandflies to human

Analysis of plasmodium vivax merozoite surface protein-1 gene sequences

The emergence of Plasmodium vivax in Dashte Moghan in northwestern Iran has become a major concern for Iranian's health officials. Knowledge of genetic make up of the P. vivax populations in this area would give us an insight into the origin of the prevalent infections and the possible routes they are introduced. This paper reports the analysis of a variable region between the two interspecies conserved blocks [ICBs] of 5 and 6 of MSP-1 gene in 18 isolates from Dashte Moghan. The results revealed that all the 18 isolates were similar to an Azari Belem-like type with 21 glutamine [Q] in the repeated residues. Our results may give a clue that the resurgent malaria in Dashteh Moghan might have primarily been introduced from Azerbaijan. However, much more molecular and epidemiological evidence are needed to confirm this hypothesis

Genetic diversity in the circumsporozoite protein gene of plasmodium falciparum from major endemic regions of Iran

Circumsporozoite protein [CSP] is one of the stage specific antigens, which is used for the development of vaccines and to investigate the genetic diversity in Plasmodium falciparum malaria parasite. Polymerase chain reaction was used for typing of CSP genes on 67 positive falciparum malaria patients from Sistan and Baluchistan Province of Iran. Three fragments were detected for CSP gene. Twenty, 38 and 4 samples showed 700, 750 and 800 bp fragments, respectively. Sequences of some samples were aligned and compared with P.falciparum csp gene in gene bank. While the falciparum malaria endemic region of Iran is classified in low to moderate group but, extensive polymorphism was observed in the samples that could be taken into account in designing malaria vaccine

Efficacy of two insecticides: Methyl carbamate and aluminium phosphide on leishmaniasis vectors in Varamin, Iran

Leishmaniosis is a prevalent tropical parasitic disease, which is caused by Leishmania protozoa. The infection can be limited in immune-competent individuals; however, in immune-compromised individuals it could proceed to chronic and ulcerative disease. The reservoirs are carnivores, and rodents and its vectors are Phlebotomus and Lutzumia. The prevalence of different spices of Phlebotomus populations and the effects of insecticides on them are investigated in Abardejhe district located in southeast of Tehran, Iran. Tablets of aluminum phosphide [3 g] and residual formulation of methyl carbamate at concentrations of 1 and 2 g/m 2 were used in rodents' burrows. Phlebotomus population was highly sensitive to both insecticides used here, and their population significantly decreased by 80% within two months after pplication of insecticides. The effects of methyl carbamate at concentrations used lasted two months longer than that of aluminum phosphide. Control and prevention of Leishmania infection depends on the habitat and behavior of vectors and reservoirs. Periodic insecticide spraying, using residual compound inside and outside residential areas, as well as breading places of sandflies, specially rodent's burrows, are shown to be very effective

Prenatal diagnosis of congenital Toxoplasmosis: validity of PCR using amniotic fluid against indirect fluorescent antibody assay in mothers

Maternal infection with Toxoplasma gondii acquired during pregnancy may result in congenital infection of developing fetuses. Up to now, decision on informed therapeutic abortion of fetuses suspected of having T gondii infection in Iran has been made based on serological findings in mothers. This might have led to unnecessary abortion of many uninfected children. We evaluated the Polymerase Chain Reaction [PCR] assay on amniotic fluids against serological findings in mothers. PCR results in this study indicated that only about one third of Indirect Fluorescent Antibody [IFA] positive mothers had passed the infection to their children. It was a sigh of relief for the majority of mothers who learned that their infants were uninfected. On the other hand it helped the mothers of infected fetuses out of an awkward predicament by making them able to make a solid decision to terminate the pregnancy or carry their children to term

Characterization of a monoclonal antiody specific for the parasite surface antigen-2 of leishmanie major

The Leishmania major Parasite surface Antigen-2 [PSA-2] is a family of glycoinositol phospholipids anchored glycoprotoins expressed in both promastigotes and amastigotes. Promastigote PSA-2 comprises three polypeptides with approximate molecular weight of 96, 80 and 50 kDa. Amastigote express a distinct but closely PSA-2 polypeptide with molecular weight of 50 kDa. In this study fusion of SP2/0 myeloma cells with immunized mice spleenocytes infected with promastigotes of L. major intraperitoneally resulted to a clone of hybridoma producing a specific antibody that only reacts with L. major parasite surface antigen [PSA-2]. This mAb showed no crossreactivity with either other Leishmania species including L. tropica, L. donovani and L. infantum or recombinant gp63. Western blot analysis of culture supernatant revealed multiple b and s with molecular weight of 50, 58, 80 and 96 kDa only in L. major

Introduction of an indirect haemagglutination test as a rapid diagnostic method in comparison with ELISA using antigen B for diagnosis of human hydatid disease

Early diagnosis of human hydatid disease by detecting the specific antibodies in patients' sera is considered as an important step in treatment of infection. But the diagnostic efficiencies of assays greatly depend on the characteristics of antigen that is used and various conditions in performance. In present study, we tried to st and ardize an indirect haemagglutination test, using antigen B for diagnosis of hydatid disease. Sera from 80 patients with surgically confirmed hydatidosis and 40 sera from healthy donors were examined. To detect the cross-reactant antibodies, 53 sera from patients with other parasitic infectious and diseases were applied in this study. IHA was performed with sheep RBC that was sensitized by various concentrations of crude antigen and antigen B. The best results were obtained by IHA with applying antigen B [10 micro g/ml] for 40 min at 37°C or 60 min at room temperature. Diagnostic value of antigen B [sensitivity 93.75%, specificity 100% and efficiency 97.12%] was significantly higher than related value of crude antigen [sensitivity 65%, specificity 100% and efficiency 83.81%] in IHA under the optimum condition. Sensitivity and specificity of ELISA using crude antigen [10 micro g/ml] were obtained 80% and 94.62%, respectively. Corresponding values of ELISA using antigen B were also obtained as 72.5% and 98.92%, respectively. It is suggested that the IHA, as a serological assay, is a valuable method with high diagnostic efficiency for serodiagnosis of hydatid disease, when is performed by purified antigen B. It is a rapid diagnostic assay with any needs neither for expensive instruments nor expert personnel so is useful for seroepidemiological studies and field trial in endemic areas

Molecular characterization of Anopheles fluviatilis species complex in the Islamic Republic of Iran

A species-specific polymerase chain reaction [PCR] assay was used to identify the species composition of the Anopheles fluviatilis complex in the Islamic Republic of Iran. All the amplified DNA samples from specimens collected from different areas yielded a fragment of 450 bp size, a PCR product corresponding to that of the species denoted as Y. The sequence data from 21 ITS2 [second internal transcribed spacer] regions were compared with those publicly available in the GenBank database and confirmed that the specimens were 100% identical to species Y of India. Species Y is presumably the same as species T that has no role in transmission of malaria in India, whereas An. fluviatilis is known as a secondary vector of malaria in the Islamic Republic of Iran

Nasopharyngeal pentastomiasis [Halzoun]: report of 3 cases

A three-member family from Tehran referred with variety of nasopharyngeal symptoms including sneezing, coughing and nasal discharge following consumption of barbecued liver [Kabab]. Thirteen worm-like nymphs of Linguatula serrata were separated from larynx, nose and gum of these patients. The adult parasites live in the nose and paranasal sinuses of dogs and other carnivores. Although this infection is rare, in population who have the habit of consuming undercooked internal organs of herbivorous animals the physicians should consider its probability

in irtro investigation on the inhbitory effects of four different species of vinca extracts on leishmania major

Vinca minor, Vinca major, Vinca variegata and Vinca rosea are different species of Vinca genus, of Apocynaceae family. In addition to ornamental applications, they are placed in the group of medicinal plants, because of the various alkaloids they possess. The purpose of this research was the investigation of the antileishmanial effects of the methanolic and crude extracts of leaves and stems of these four plant species on L.major promastigotes in vitro. The results showed that the methanolic extracts of all four species induced a significant decrease in the number of L.major promastigotes in different degrees and an increase in concentration and exposure time would destroy the parasites completely