Journal of School of Public Health and

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Visceral leishmaniasis (VL) has been found as an endemic disease in some areas in northwest and south parts of Iran during recent two decades. The species of the Leishmania has been characterized as L.infantum and the main sources of human infection in the endemic areas is dog. The majority of kala-azar cases are found among children in the age group of 1-4 years. As the delay in diagnosis and treatment of kala-azar cause high mortality in the patients, serological surveillance, using direct agglutination test (DAT), and treatment of seropositive cases who have clinical symptoms are carrying out with cooperation of Provincial Health Services in the endemic foci of Ardebil and East Azerbaijan Provinces in the northwest and Bushehr in the south parts of Iran since 1997.
DAT Leishmania antigen is made in the Protozoology Unit of the School of Public Health with the strain of L.infantum isolated from an infected dog in Iran. The finger prick blood samples are collected by trained Health Workers (Behwarz) from suspected kala-azar patients in the Rural Health Houses. The collected samples are transferred to the near district kala-azar laboratory and tested (using DAT) by the trained technicians. The sero -positive patients are referred to pediatricains or trained general physicians in district hospital or health center for clinical examination and treatment of the seropositive cases (DAT titers of 1:3200 or higher) with kala-azar clinical symptos. For the treatment, usually, meglomine antimongtc (Glucantime) is used via intra-muscular in dosss of 20mg/kg/day for 20 days.
In kala-azar case finding in the serological surveyed areas of Ardebil, East-Azerbaijan and Bushehr provinces during 1997-2001, altogether, 19693 blood samples were collected from suspected kala-azar patients and tested by DAT. Totally , 1274 cases (6.74%) were seropositive in titers of 1:3200 or higher. In the clinical examination of seropositive cases 690 patients (54.16% of sero-positive cases and 3.50% of the total samples) had kala-azar clinical symptoms and therefore they were treated.
This seroloeical surveillance of kala-azar in the endemic area as have detected high number of kala-azar patients among children up to 12 years old, who were subsequently treated on time . This ptoject has prevented the mortality of the disease in the studied areas.

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Toxoplasmosis is a common disease caused by the protozoal parasite Toxoplasma gondii. Most human cases occur by 1) eating raw or undercooked meat containing T. gondii tissue cysts. 2) ingestion of oocysts from soil and 3) vertical transmission through the placenta. Immunocompetent adults are usually asymptomatic or present with self-limited fever and lymphadenopathy. Infection acquired during pregnancy can be transmitted to the fetus and may cause mental retardation, blindness, epilepsy, and abortion. In this cross-sectional study we determined the prevalence of toxoplasma infection in residents of Meshkin-Shahr in 2001-2002. We applied a cluster sampling method to family health files in local health centers to recruit a total of 909 individuals for this study. All family members were requested to complete the study&aposs questionnaire. For each individual, two separate blood samples were collected and placed in microhematocrite tubes. Titers of anti-toxoplasma antibodies were measured by IFA with levels in excess of 1/20 taken as positive. Questionnaire information and examination results were analyzed by the SPSS software package using chi-square tests. Results of this survey show the overall prevalence of toxoplasma infection to be around 18.3%. Figures for men and women were 19.7% and 17.2%, respectively but the difference was not statistically significant. Prevalence rates show a significant association with age (greater prevalence in higher age groups) and the type of meat consumed (P<0.05). But there were no significant associations with education level, occupation-keeping cats in the house, ingestion of meat and history of previous abortions. The most frequently occurring antibody titer was 1/20 (6.9%). and frequency decreased with higher antibody titers.
This study shows that toxoplasma infection is not as common in Meshkin-Shahr as in some parts of the country, with more than 80% of individuals being seronegative. Because of the low immunity level and the risk of vertical transmission, educating the public in prevention methods could be very important.

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This study is a retrospective review of the epidemiology of kala-azar, using cases reported to health facilities in Ardebil province over a period of 5 years: from the beginning of 1997 until the end of 2001.
The total number of cases was 602, but as the assessment was only possible in hospitalized patients, the questionnaires were completed for 293 cases with detailed records in 5 hospitals (3 in Ardebil and two in Meshkin-shahr and Germi). Another 76 cases were also excluded (due to incomplete records) and in the end 217 questionnaires were analyzed by the Epi-info 2002 package.
Compared to the total number of cases (1051) reported in a similar study over the years 1985-90, the reduction in total number of cases (602) may be due to reduced incidence rate in Meshkin-Shahr and/or underreporting. Although the total number is down, the number of new cases in Germi and Bilehsavar districts has increased. In our study the overall male to female ratio was 1.4 to 1 this ratio was 1.5 to 1 in hospitalized patients. 74% (160) of cases were under 2 years of age and 93% (201) under 5. Only 1.8% (4) of patients were over the age of 10 years. Though the majority of the patients were seen in Ardabil city, most had been referred from Germi district. 87.5% were village dwellers, mostly referred on cold months of the year the number of referrals was lowest during summer. The most frequently used diagnostic test (especially in recent years) was DAT, positive in 77% of tested cases (titers/1600). We also performed BMA on 30 patients, of which 87% (26) were positive. 58%) of the subjects had been diagnosed in about a month after the disease onset and 93% within 4 months. The most common clinical features were anemia, fever, anorexia, splenomegaly (all present in >90% of cases) other manifestations were malnutrition, cough, hepatomegaly, diarrhea, vomiting, Iymphadenopathy, hemorrhage, jaundice, edema and ascitis. In 6 cases (3%) the chief complaint was febrile convulsion. The most common laboratory findings were anemia (mean Hb - 8.96 mg/dl), elevated ESR (highest level being 140mm/l hour), thrombocytopenia and neutropenia. Liver enzymes were elevated in one third of the patients. 96% of the cases treated with Glucantime the remaining 4% were relapse cases or treatment failures and had been given Amphotricin-B. 95.8% (211 cases) had been discharged in good general condition 3% (6) had died. The exact causes of death were not mentioned in hospital records.
According to these findings, Kala-azar is a major health problem in Ardebil province, especially within the rural population and specifically in Germi and Bilehsavar districts. It requires more intense control and more medical facilities alongside education programs for health workers and general population.

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The purpose of this study was to determine toxoplasmosis seropositivity rates and to record ascending serologic titers in high-school girls. We also searched for possible risk factors. This descriptive-analytic study used 414 serum specimens collected from high school girls in six different regions of Esfahan city through cluster random sampling. The sera were taken from the tip of the finger by hematocrite microtubes. Samples were studied by the Indirect Immunofluorescent Assay Test(IFAT) for estimation of serum titers. Data were analyzed using chi-square (X²) and t tests. The overall seropositivity rate was 18.4% in 14-19 year-old girls and this rate increased with age. IFAT titers in 98% of the positive samples were at 1:100 the remaining 2% showed values above this threshold. For the latter group, we performed another titration test to determine exact titers .The supposed risk factors were age, place of residence, education level (parents and student), consumption of undercooked meat and raw liver, occupation and parent income, and exposure to contaminated material from cats and fowls. The highest positivity rates (27.5%) were recorded in District 1, while the lowest rates (14.5%) were seen in District 5. Significant relationships were found between seropositivity and exposure to cats and fowls.(P>0/05). No significant relationship was observed with the other factors. There was a low level of knowledge about toxoplasmosis and the relevant risk factors: only 2.4% of the subjects were relatively well-informed on this subject. There were not any acute cases. Toxoplasma infection is very important because of its socioeconomic implications, so control measures seem crucial. All seronegative women should be know about this infection and its transmission routes. Education is the most important element in prevention and must be promoted via the mass media, other education systems and the health managers. The studied group in this survey were girls at the usual age of marriage or younger, and it is recommended that health system managers continue to offer education on practices that help prevent infectious diseases in general and toxoplasmosis in particular.

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Background and Aim: In this study, we compared a latex agglutination test (KatexR) with the direct agglutination test (DAT) for the diagnosis of human visceral leishmaniasis (HVL) in an area with a high prevalence of zoonotic visceral leishmaniasis (ZVL) in Iran.
Materials and Methods: The validity of the KatexR (Kalon Biological Ltd, England) for detection of urinary Leishmania antigens in HVL was compared with standard direct agglutination test on 92 visceral leishmaniasis patients (29 were acute and 65 were chronic phase) and 90 healthy controls over a period of one year.
Results: We detected a sensitivity of 82.7% and a specificity of 98.9% in the acute phase. As for the chronic phase, the test showed a sensitivity of 6.15% and a specificity of 98.9% at a 1:3200 cut-off titer. Concordance between KatexR and DAT was 94 .9% in the acute phase and 59. 3% in the chronic phase. As the rapid, noninvasive Katex does not require much expertise or elaborate equipment.
Conclusion: It can be used for screening and diagnosis of human visceral leishmaniasis in remote endemic foci.

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Background and Aim: Visceral leishmaniasis (kala-azar), the most dangerous form of leishmaniasis, is endemic in some parts of Iran, e.g. Ardabil, Fars, East Azerbaijan, Bushehr and Qom provinces. In recent years, the incidence of VL has increased in the Nourabad-Mamassani district in Fars Province. This study was carried out to detect VL vectors and infection rates in this region over the 2003-2004 period.
Material and Methods: Sand flies were captured in the selected villages by means of sticky traps, aspirators and CDC miniature light traps. Heads and distal abdominal segments were used for species identification and other body parts were used for DNA extraction. We employed a semi-nested PCR technique to detect Leishmania, with specific kDNA primers (LIN R4 - LIN 17 – LIN19). Some specimens were dissected for leptomonad infection.
Results: A total of 12688 sand flies were collected. Phlebotomus (Paraphlebotomus) alexandri was the second most prevalent species (17.34%). The anthropophilic index of this species was 32.5%. Five cases (4.17%) of L. infantum infection were detected among the 120 P. alexandri examined by PCR method. We also observed two cases of leptomonad infection among the 112 dissected specimens.
Conclusion: High prevalence rates and anthropophilic index of P. alexandri plus its natural infection with L. infantum provide enough evidence to implicate this species as the main vector species of VL in the region and the second proven kala-azar vector in Iran. Besides, the Mahoor-Milaty district of Noorabad-Mamassani was identified as a new endemic focus.

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Background and Aim: Toxoplasma gondii is a protozoan parasite infecting humans and warm-blooded animals. Toxoplasmosis in pregnancy could cause neurologic disorders in the fetus. In immunocompromised hosts, the infection can be reactivated with life-threatening consequences. Detection of the parasite or its components would constitute a better definition of acute infection.

Material and Methods: To detect T. gondii antigen and DNA, twenty serum samples in the acute phase of infection were tested. Polyclonal antibodies were isolated from immunized rabbits and SDS-page and immunoblotting were performed. Also, PCR was done with amplification of the B₁ gene with two primers.

Results: In one patient T. gondii antigen band with a molecular weight of 30 kDa was detected. Parasitemia was detected in the same patient and the 570 bp amplified DNA fragment was isolated. Others had negative results in both immunoblotting and PCR. The patient with positive results had been infected accidentally with a rather virulent C56 strain in the laboratory.

Conclusion: The results indicate that antigen and DNA of T. gondii can be detected during the short acute phase of infection.

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Background and Aim: Leishmaniasis is a parasitic disease transmitted by sand-flies. Seventeen out of the 30 Iranian provinces are contaminated. Although many therapeutic modalities have been suggested, no definite treatment for this widespread infection is available. Many drugs currently used for cutaneous leishmaniasis have their side effects and create complications-drug resistance, etc. This has prompted researchers to get interested in drugs of a natural origin. The aim of this study was to investigate and compare the efficacies of an extract of propolis and systemic glucantime against cutaneous leishmaniasis in Balb/c mice.

Materials and Methods: Groups of inbred, female Balb/c mice, aged 4-6 weeks, were inoculated with 1.6 × 10 6 viable stationary-phase promastigotes in the base of tail. The mice were provided by Iran Pasteur Institute, Tehran and were randomized into three groups of 9 mice each. Groups 1 to 3 were treated with pure 70% ethanol, systemic glucantime, or the propolis hydroalcoholic extract for a 6-week period the 4th group served as control. The statistical tests including student t-test were used for data analysis.

Results: Overall, 27 mice were studied. We did not observe any statistically significant difference among the groups with regard to the size of the ulcer at the beginning (P>0.01). At the end f the period, there was a significant difference with regard to reduction of ulcer size between the propolis extract and glucantime groups in favor of the propolis extract (P= 0.001). Further analysis of the data showed that the largest reduction in the ulcer size was caused by the propolis extract, as compared with the control, glucantime and alcohol groups.

Conclusion: Our results suggest that the propolis extract is more effective in the treatment of cutaneous leishmaniasis in mice. We suggest that the efficacy of this extract alone or in combination with drugs be assessed against human cutaneous leishmaniasis, using randomized clinical trials.

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Background and Aim: Visceral leishmaniasis is a systemic parasitic disease with a high fatality rate in under-5-year-old children. The disease is endemic in some parts of Iran, particularly in the north-west region. In 2001 a visceral leishmaniasis (VL) surveillance system was established for children aged ≤ 12 years in the primary health system in Meshkin-Shahr District, Ardebil Province, situated in the north-west of Islamic Republic of Iran.

Materials and Methods: All cases with clinical signs and symptoms of VL and confirmed positive by the direct agglutination test (DAT) were referred for physical examination and treatment.

Results: The mean annual incidence of VL decreased significantly from 1.88 per 1000 children before (1985-2000), to 0.77 per 1000 child population after (2001-07), the intervention. In the control area with no surveillance, it increased from 0.11 to 0.23 per 1000.

Conclusion: Early detection of VL using serological tests and timely treatment of cases can decrease the mortality and morbidity rates of VL in endemic areas.

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Background and Aim: Problems related to conventional Fenton oxidation, including neccesity of having a low pH and production of considerable amounts of sludge, have prompted researchers to consider chelating agents to improve the pH operating range and iron nano-oxide particles to reduce excess sludge. The main objective of this study was to remove pyrene from contaminated soils by a modified Fenton oxidation method  at neutral pH.

Materials and Methods: Experiments were conducted using various concentrations of H2O2 (0-500 mM), iron nano-oxides (0-60 mM), reaction times (0.5-24 hours) and several chelating agents, namely, sodium pyrophosphate, ethylene diamine tetra-acetic acid, sodium citrate, fulvic and humic acids, to eliminate pyrene from soil (concentrations of 100-500 mg/kg).

Results: The efficiency of removal of pyrene at an initial concentration of 100 mg/kg was 99 % at the following reaction conditions: H2O2 and iron nano-oxide concentrations of 300 mM and 30 mM, respectively pH=3 and a reaction time of 6 hours. The initial pyrene concentration of 100 mg/kg decreased to 7 mg/kg at optimum conditions using sodium pyrophosphate as the chelating agent at pH 7.    

Conclusion: The modified Fenton oxidation method, using iron nano-oxide at optimum conditions as defined in this research, is an efficient alternative for chemical remediation or pre-treatment of soils contaminated wih pyrene at neutral pH. 

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Background and Aim: Considering the ongoing national malaria elimination program in Iran, establishing a bank of human Plasmodium genes and proteins can be very useful for research purposes. This study was conducted to collect some of the native isolates of human Plasmodia from endemic areas in the country.

Materials and Methods: A 2ml vein-punctured blood sample was prepared from each confirmed malaria case. The samples were dispensed in EDTA pre-dosed tubes and cryopreserved for further tests. Moreover, relevant Geimsa-stained thick and thin blood smears were kept in a safe place. Tests for genetic indicators of MSP-1 was performed for each of the P. vivax samples with the RFLP-PCR techniques. In addition, an in vivo drug sensitivity test was performed for each P. falciparum case. Collecting and cryopreserving samples will continue.

Results: A total of 131 samples, including 109, 19 and 3 P. vivax, P. falciparum and mixed samples, respectively, were preserved with relevant data such as species, parasitaemia and nationality of the donor. MSP-1 gene classification resulted in three different haplotypes including Hap.1, Hap.2 and Hap.3 with frequencies of 20.6%, 41.2% and 38.2%, respectively. The In vivo drug sensitivity tests on P. facilparum isolates showed that all of the isolates were sensitive to the current drug of choice, namely, a combination of artesunate and fansidar.

Conclusion: This study resulted in the preservation of considerable amounts of P. vivax and P. falciparum samples for further relevant studies and research purposes.