Seroepidemiology of herpes simplex virus type 1 and 2 in northern iran.

BACKGROUND: 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. METHODS: 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. RESULTS: HSV-1 and HSV-2 IgG antibodies were 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). CONCLUSION: Our findings were in agreement with prior studies in which HSV-1 infections was more prevalent than HSV-2 and seropositivity increased with age.

Inhibition of Murine Systemic Leishmaniasis by Acetyl Salicylic Acid via Nitric Oxide Immunomodulation.

BACKGROUND: 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. METHODS: 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. RESULTS: ASA increased NO production in plasma of both naïve 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. CONCLUSIONS: 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.

Aflatoxin M1 in Pasteurized Milk in Babol city, Mazandaran Province, Iran.

BACKGROUND: Aflatoxin M(1) (AFM(1)) is the metabolite of aflatoxin B1 (AFB(1)) and is found in milk when lactating animals are fed with contaminated feedstuff. The presence of AFM(1) 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 AFM(1) in subsidized pasteurized milk in Babol, Mazandaran Province, Iran. METHODS: 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 µl of skimmed milk were tested for AFM(1) contamination by competitive ELISA. RESULTS: All the samples (100%) exhibited contamination with AFM(1). The contamination levels means in January, February, and March were 227.85, 229.64, and 233.1ng/l, respectively. The amount of AFM(1) in all the samples were above 50ng/l, the threshold set by the European community regulations. CONCLUSION: Monitoring of AFM(1) 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 AFM(1) 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.

BACKGROUND: 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 negative by direct smear and culture, with PCR technique. METHODS: 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 RESULTS: Out of 161 negative samples by direct smear and culture, seven samples (4.3%) were positive by PCR technique. CONCLUSION: 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.

Leishmania major: common antigen responsible for induction of delayed-type hypersensitivity response in guinea pigs.

The cellular response to Leishmania major (L. major) was evaluated in vivo by delayed-type hypersensitivity (DTH) skin test reaction using leishmanin as antigen. Our previous study had shown the development of species-specific DTH reaction in sensitized guinea pigs by application of a single purified antigen from promastigotes and filtered culture supernatants of L. major. This study has shown that purified antigen is common in both stages of the life cycle and filtered culture supernatant of L. major. The common antigen was purified and analyzed from soluble Leishmania antigen (SLA) of amastigotes, promastigotes, and filtered culture supernatant of L. major by specific monoclonal antibody coupled to sepharose-4B. The purified antigen, which gave a single band of 56 kDa on sodium dodecyl sulfate polyacrylamide gel (SDS-PAGE) electrophoresis, elicited DTH response in guinea pigs sensitized with L. major. It was almost of the same degree as that produced by whole SLA. These results show that DTH inducer antigen is present in both stages of the life cycle and filtered culture supernatant of L. major.

Analysis of specific IgE and IgG subclass antibodies for diagnosis of Echinococcus granulosus.

The potential roles of specific antibodies of different immunoglobulin G (IgG) subclasses and IgE in serological diagnosis of cystic echinococcosis (CE) were investigated by an enzyme linked immunosorbent assay (ELISA) based on Antigen 5 (Ag5). Presence of IgG1 was demonstrated in all sera from 58 patients with CE. The most discriminatory and specific antibodies found in this study belonged to IgG4 and IgE. Only one false-positive reaction was observed with IgG4 and no IgE cross-reactivity occurred with 40 sera from healthy controls. In 36 sera from patients infected with parasites other than CE two false-positive reactions with IgG4 were observed but none occurred with IgE. In immunoblotting, it was shown that IgG1 subclass was responsible for cross-reactivity of human antibodies that reacted with a 38 kDa subunit of Ag5. IgG4 and IgE antibodies could not recognize the 38 kDa subunit and under non-reducing conditions reacted with the 57 kDa subunit without any cross-reactivity to other parasites. The results demonstrated that IgG4 and IgE are the most important antibodies for serological diagnosis of hydatid cyst in an Ag5 based immunoassay system.

Nitric oxide induction as a novel immunoepidemiological target in malaria-infected patients from endemic areas of the Islamic Republic of Iran.

OBJECTIVE: Malaria has been prevalent for a long time in Iran and continues to be a health problem despite substantial control programs. In addition to numerous cytokines, nitric oxide (NO) is thought to be a key molecule and a novel target of malaria immunopathology. MATERIAL AND METHODS: The objective of this research was to measure reactive nitrogen intermediates (RNI) as stable metabolites of NO induction in plasma of malaria-infected patients in Iran. In this study, 235 blood samples from malaria patients and 80 blood samples from healthy controls were randomly collected from different malarial endemic provinces of Iran, located in southeastern (Sistan & Balouchestan, Hormozgan, Kerman) and northwestern (Ardabil) areas. The involvement of NO in malaria patients has been investigated by statistical analysis of RNI values. Griess micro assay (GMA) was used during Plasmodium vivax, P. falciparum and mixed infections, in order to evaluate whether RNI changes are related to the provincial areas, parasite strains, clinical symptoms and age and gender parameters. RESULTS: The results showed a significant increase of RNI level in malaria patients compared with the control groups of Ardabil (p<0.01), Sistan & Balouchestan, Hormozgan and Kerman (p<0.001) provinces. The level of RNI was higher in mixed plasmodial infection than in single infection. CONCLUSIONS: The high level of RNI was dependent on the type of infection, the plasmodia strain, the clinical symptoms, the age groups and the endemic provinces. Although, this study did not clarify the pathogenic and/or protective role of NO in malaria, our findings provide a novel immunoepidemiological aspect of basal NO production in patients with malaria in endemic areas in Iran.

Leishmania major: species specific delayed hypersensitivity reaction induced by exogenous secreted antigen in the guinea pig.

The cellular response to Leishmania major (L. major) is usually evaluated in vivo by the delayed-type-hypersensitivity (DTH) test using leishmanin. Leishmanin can give false-positive reactions in areas where there is a background of leishmaniasis. In a previous study, it was shown that a 56 kDa antigen purified from promastigote and culture supernatant of L. major induce strong DTH reactions in sensitized guinea pigs. In this study, the species-specificity of this antigen was further investigated. Three groups of guinea pigs were sensitized with L. major, L. tropica, and L. infantum and both flanks of sensitized animal were injected intradermally with purified 56 kDa antigen or soluble leishmania antigen (SLA). The extent of indurations were measured after 24, 48, and 72 h. In animals which were sensitized with three species of leishmania, only those immunized with L. major showed skin reactions to purified antigen by an increase in skin thickness. Since complex antigen mixtures such as SLA and leishmanin show cross-reactivity and can be non-specific, the result obtained here suggest that 56 kDa antigen may be a useful diagnostic tool for species specific diagnosis in field studies of leishmaniasis.

Leishmanin skin test in guinea pig with a single purified protein of Leishmania major.

A series of hybridomas was produced by fusion of SP2/0 myeloma cells with spleen cells of mice immunized with Leishmania major (L. major). The reactivity of secreted monoclonal antibodies (mAbs) was evaluated against available leishmanin antigen by enzyme linked immunosorbent assay. Only one hybridoma designated as 7F9 secreted IgG1 mAb which was shown to be reactive with leishmanin. This mAb was further tested against four species of Leishmania (L. donovani, L. tropica, L. infantum, L. major) and a recombinant gp63. Among the four species tested it was shown to be only reactive with promastigotes of L. major. The antigen recognized by this mAb was purified and analyzed from both sonicated and supernatant cultures of L. major by immunoaffinity chromatography and reverse phase high performance liquid chromatography. The purified antigen, which gave a single band of 56kDa on sodium dodecyl sulfate polyacrylamide gel electrophoresis elicited a strong delayed-type hypersensitivity (DTH) reaction in guinea pigs sensitized with L. major. It was almost of the same degree as that produced by leishmanin. These results suggest that an L. major-specific antigen is an alternative as a specific diagnostic skin test reagent, which could lead to a better understanding of the mechanism of DTH in L. major.

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.

rDNA-ITS2 based species-diagnostic polymerase chain reaction assay for identification of sibling species of Anopheles fluviatilis in Iran.

A species-specific polymerase chain reaction (PCR) assay using primers already designed, based on differences in the nucleotides of the second internal transcribed spacer (ITS2), was used to identify the species composition of the Anopheles fluviatilis complex in Iran. All the amplified DNA samples obtained from specimens collected from different areas using different collection methods yielded to a fragment of 450 bp size, a PCR product corresponding to the species denoted as Y. Some 21 ITS2 region of Iranian specimens were sequenced and compared with the already published sequence data of species Y from India. The sequence data of the Iranian specimens were 100% identical to that of the Indian specimens, and hence confirmed the PCR assay results. Species Y is presumably species T in India, which has no role in the transmission of malaria, whereas mosquitos of An. fluviatilis are known as a secondary vector in Iran. This conflict will remain to be solved by further biological and molecular studies.

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

Effects of viral hemorrhagic fever inactivation methods on the performance of rapid diagnostic tests for Plasmodium falciparum.

Blood samples from patients with viral hemorrhagic fever (VHF) pose a serious risk to laboratory workers. Current contingency plans for VHF samples recommend the use of heat, gamma-irradiation, or Triton X-100 to inactivate samples before handling. Malaria is the most important alternative diagnosis to be excluded in cases of suspected VHF. Interpretation of malaria smears using samples inactivated with these methods is problematic because morphology is altered. The objective of this study was to assess the impact of different inactivation methods on the performance of rapid diagnostic tests for Plasmodium falciparum. Triton X-100 and gamma-irradiation of samples preserved detection. The impact of Triton X-100 inactivation was also "blindly" evaluated using 100 blood samples from febrile travelers. Triton X-100 inactivation of samples did not significantly affect the performance of these tests. This may represent a useful strategy for excluding the diagnosis of falciparum malaria in cases of suspected VHF.

[Toxoplasmosis in Iran. Results of a seroepidemiological study]. / Toxoplasmose en Iran. Résultats d'une étude séroépidémiologique.

A toxoplasmosis seroepidemiological survey was effected with 13,018 sera collected by stratified cluster random sampling method from 12 provinces in Iran. The samples were studied by indirect immunofluorescent assay (IFA) for the presence of Toxoplasma. In this study, 52.6% of the subjects were male and the remaining 47.4% were female. Anti-Toxoplasma antibody was detected in a total of 51.8% of the samples with no significant difference between male and female affected subjects. The distribution of the infected samples was also investigated in various age groups, the level of infection to Toxoplasma increasing from childhood, culminating to 30 years of age and gradually declining from there after. Between the various age groups, the 10-19 years old demonstrated a 50% increase in relative risk to the infection with high antibody titer. Within the provinces under study, the highest relative frequency of Toxoplasma antibody titer was indicated in Mazandaran province (20.5%), while the lowest frequency was detected in Hormozgan (2.9%). In general, there was a decrease in the number of infected samples from humid areas in north to dry provinces in south of Iran. In the clinical symptoms study, no significant difference between male and female patients was demonstrated. According to the type of clinical manifestation, lymphadenopathy and central nervous system symptoms (encephalitis) were respectively the most and the least frequent manifestations.