[Comparison between the effects of ethanolic extract of Artemisia annua and chloroquine on Plasmodium berghei in white mice]

Considering the importance of using native and traditional herbal medicines in the treatment of malaria, we made a comparison between the effects of an ethanolic extract of Artemisia annua and chloroquine on Plasmodium berghei in Sourian mice. In this study 50 Sourian mice were divided into 10 groups, each group consisted of five mice. The evaluation was done according to the Peters test. Nine of ten groups were infected with P. berghei. The first 6 groups were given extracts of Artemisia annua at different concentrations. The seventh and eighth groups received chloroquine and placebo, respectively. The ninth group received no treatment [control group].The tenth group was not infected with Plasmodium berghei and did not receive any treatment and was used for determination of accidental mortality of the mice. In each group, the levels of parasitaemia were determined on the 4[th] and 7[th] days, and compared with those of the control group. To find the most effective concentration, the test was repeated with 55 mice divided into 11 groups using limited spectrum of drug concentrations. We used SPSS software and T-test for data analysis. The results indicated that Artemisia annua extract at the concentrations of 1100 and 1300 mg/kg significantly decreased P. berghei parasitaemia in the infected mice, but at the concentration of 1100 mg/kg Artemisia annua had less toxicity [P<0.05]. According to the results of this study Artemisia annua at the concentration of 1100 mg/kg showed considerable effect on P. berghei

[Entomological research and its specific course in Iran from 1935 to 2008]

Malaria remains an important vector-borne disease globally and is a threat for human life. Forty percent of the world's populations who are living in low-income countries are at risk of malaria. The disease exists in Iran and caused economic and social damages. As result of malaria control program that has been done during the past years, the disease is eliminated from the most parts of the country, so that it is only reporting from a small part in these years. During this study, all available papers, books and thesises were reviewed and articles from Iranmedex, DIS and PubMed databanks were also used. Furthermore the related reports from different sources were noted. The extensive studies have important information about malaria vectors. In this study the data about malaria vectors and related training courses are listed. During this study the related papers, Books and thesises which have been reviewed. Although efforts, surveillance system, diagnostic and treatment facilities, as well as knowledge and attitude of peoples regarding to health behavior are improved nowadays, there are significant improvements about decreasing the malaria cases. Risk of the disease exists because of population exchange and asymptomatic cases. The malaria can be studied with both public health and economical aspects. This paper represents entomological studies of malaria during 1935 by the end of 2008. our study revealed that, based on recent malaria national program, the authorities should make an emphasis on vector control monitoring, resistance management, malaria evaluation and because of weak supervision on all malaria operation at stage of elimination of malaria, accurate and careful suppersional require to reach the objective and goal of elimination

Comparison of microscopical examination and semi-nested multiplex polymerase chain reaction in diagnosis of Plasmodium Falciparm and P. vivax

We compared light microscopy examination and a semi-nested multiplex PCR [SnM-PCR] assay in endemic areas of the Islamic Republic of Iran. A total of 68 individuals with malaria-positive and suspected malaria symptoms were included in the study. Giemsa-stained thick blood films were examined under a light microscope for malaria parasites in 100 and 200 fields. DNA was extracted from blood samples and SnM-PCR based on the amplification of the small subunit ribosomal RNA [ssrRNA] gene sequences was applied. Microscopical examination showed that 48.5% [33.8% P. vivax and 14.7% P.falciparum] and 50% [35.3% P. vivax and 14.7% P.falciparum] of the samples were positive in 100 and 200 fields respectively. SnM-PCR showed the same results as the 200 field microscopy

Sequence analysis of different domains of plasmodium vivax apical membrane antigen [PvAMA-1 gene] locus in Iran

Plasmodium vivax is responsible for approximately 80 million malaria cases in the world. Apical membrane antigen1 [AMA-1] is a type I integral membrane protein present in all Plasmodium species. AMA-1 interferes in critical steps of invasion of human hepatocytes by sporozoites and red blood cells by merozoites and is one of the most immunodominant antigens for eliciting a protective immune response in human. It is considered as a promising antigen for inclusion in a vaccine against P. vivax. Since more knowledge is needed to lighten the scope of such antigen we compared genetic variation in P. vivax AMA-1from an Iranian isolate with those reported from some of the other malarious countries so far. P. vivax genomic DNA was extracted from the whole blood of an Iranian patient with patent P. vivax infection. The nucleotide sequence for 446 amino acid [AA] residues [42-488 of PvAMA-1] was amplified by PCR and cloned in pUC19 vector for sequencing. Sequence analysis of the antigen showed a high degree of identity [99%] with strong homology to the PvAMA-1 gene of P. vivax S3 and SKO814 isolates from India and Korea [Asian isolates] respectively, and 96% similarity with P. vivax Sal-1 AMA-1 gene from El Salvador. We cloned and characterized three domains of PvAMA-1 gene from an Iranian patient. Predicted protein sequence of this gene showed some discrepancies in corresponding protein in comparing with similar genes reported from other malarious countries

parasitological and serological study in malaria suspected patients in Hormozgan Province, Southeastern Iran

Presence of malaria immune factors induced by erythrocytic stages is widely used as an epidemiological approach to diagnose the infection mainly to distinguish the current, recent and past infections. This study was performed to find out the status of malaria, using microscopical and serological [IFA] methods in Bandar-Abbas and Minab, two malarious districts in Hormozgan Province of Iran. 408 patients with suspected malaria symptoms were enrolled. Conventional microscopic examination and serological IFA test were employed for diagnosis of malaria. The rates of agreement between microscopical and serological diagnosis were analyzed by Kappa test. 17.9% and 1.7% of the samples were microscopically diagnosed as P. vivax and P. falciparum, respectively. On the other hand, the serum samples were sero-positive with P. vivax and P. falciparum antigens in 54.2% and 32.1% of the samples, respectively. Serological IFA method could mainly determine the past history of malaria infection, but it was not helpful in detection of current infections. Moreover, there was no significant agreement between microscopical and serological [IFA] method0s in diagnosis of malaria

Molecular monitoring of plasmodium vivax infection after radical treatment in southeastern Iran

The aim was to evaluate the relapse risk of vivax malaria in patients who received radical treatment in Hormozgan Province, a malarious area located on southeast of Iran. A total of 95 symptomatic vivax malaria infected patients were enrolled in urban health centers of Bandar-Abbas, Minab, Bandar-Jask and Bashagard districts of Hormozgan Province, southeast of Iran from January 2008 to March 2009 for consideration as a case-series study. DNA was extracted from parasite infected whole blood samples. A polymorphic region of Plasmodium vivax merozoite surface protein 1 [pvMSP1] was selected and a PCR method was employed for all the samples to amplify the specific variable gene fragment. The obtained fragments in primary and secondary samples were sequenced. Both nucleotide and amino acid sequences of the samples were investigated for returned patients. 3.2% of the patients experienced a second attack between 83-199 days after the initial episode of infection. Alignment of nucleotide and their deduced amino acid sequences between pair sequences of primary and secondary isolates revealed 8 and 6 dissimilarities respectively for the first case, and 9 and 7 dissimilarities for the second case. Although microscopical examination of recurrent thick blood smear of the third patient confirmed new P. vivax infection, the venous blood sample was accidentally missed. Sequencing results of primary and returned isolates 1P, 1S, 2P, 2S and 3P in this study showed an identity with BP13, T117, BP13, TC28 and Chesson genotypes respectively. The returned [secondary] isolates may account to be for the sake of reinfection

Residual effects of deltamthrin WG 25% as a new formulation on different surfaces against anopheles stephensi, in Southeastern Iran

Indoor residual spraying [IRS] is functioned as national interventions against malaria in southeastern foci of Iran and deltamethrin WP one of the insecticides have been used since past decade. In this study, the residual activity of the wettable granule [WG] was studied on different surfaces in hut scale trial against Anopheles stephensi in Iranshahr district, southeastern Iran. Three dosages of 25, 40 and 50 mg a.i./m2 of deltamethrin WG 25% formulation were applied on plaster, cement, mud, and wooden surfaces using Hudson[Registered] X-pert compression sprayer having 10 litters capacity. The residual effects of deltamethrin WG 25% on different surfaces was assessed based on reduction of mortality An. stepehnsi from 100% to about 70%. At 25, 40 and 50 mg a.i./m2 the WG formulation of deltamethrin had a bioefficacy for about 2, 3 and 4 months respectively. There was an expectable fluctuation in mortality of An. stephensi at different sprayed surfaces as well as dosages. The proposed 50 mg/m2 WG is the longest activity for up to 4 months which needs to be applied for two spraying cycles per year at the climatically condition of southwestern Iran

Partial sequence analysis of merozoite surface protein-3 alpha gene in plasmodium vivax isolates from malarious areas of Iran

Approximately 85-90% of malaria infections in Iran are attributed to plasmodium vivax, while little is known about the genetics of the parasite and its strain types in this region. This study was designed and performed for describing genetic characteristics of plasmodium vivax population of Iran based on the merzoite surface protein-3 alpha gene sequence. Through a descriptive study we analyzed partial P. viva merozoite surface protein-3 alpha gene sequences from 17 clinical P. vivax isolates collected from malarious areas of Iran. Genomic DNA was extracted by Q1 Aamp DNA blood mini kit, amplified through nested PCR for a partial nucleotide sequence of PvMSP-3alpha gene in P. vivax. PCR-amplified products were sequenced with an ABI Prism Perkin-Elmer 310 sequencer machine and data were analyzed with clustal W software. Analysis of PVMSP-3alpha gene sequence demonstrated extensive polymorphisms, but the sequence identity between isolates of same types was relatively high. We identified specific insertions and deletions for the types A, B and C variants of vivax in our isolates. In phylogenetic comparison of geographically separated isolates, there was not a significant geographical branching of the parasite populations. The highly polymorphic nature of isolates suggests that more investigations of the PvMSP-3alpha gene are needed to explore its vaccine potential

Variation of the chloroquine resistance transporter [crt] gene in chloroquine-resistant and chloroquine-sensitive plasmodium berghei

The emergence and spread of chloroquine resistant Plasmodium falciparum in the world stimulated some investigators to consider different aspects of chloroquine resistance in human and rodent Plasmodia. Using animal Plasmodia, particularly primate and rodent Plasmodia can be useful model for human Plasmodia studies. In this study we have tried to consider and compare the sequence of chloroquine resistance transporter [crt] gene among chloroquine-resistant and chloroquine-sensitive strains of Plasmodium berghei. This experimental study was performed at the Malaria Laboratory of School of public health. DNA was extracted from two strains of P. berghei which their resistance and sensitivity had been demonstrated in mice with treatment by chloroquine. By using specific primer for crt gene some parts of this gene were amplified by PCR, and obtained fragments were then sequenced and compared. There were considerable differences in crt gene between two strains. Sequenced 1212 bp of crt gene fragment in the two strains showed 43 differences at nucleotides level and 16 differences in presumed coding amino acids. crt can be addressed as a considerable gene which involves in induction of resistance to chloroquine in P. berghei, as P. falciparum. The results increased such a promise that considering crt gene in chloroquine-sensitive and chloroquine-resistant P. berghei can prepare suitable and helpful fields for more understanding the molecular aspects of chloroquine-resistance in Plasmodia and reversing the effectiveness of 4-aminoquinolines particularly chloroquine for treatment of drug resistant Plasmodia

Diversity of merozoite surface protein-3beta gene of plasmodium vivax isolates from Iran

Plasmodium vivax malaria accounts for approximately 88% of malaria cases in Iran. There is limited information on genetic diversity of P. vivax in the country and a need to develop and apply an effective vaccine against the disease is necessary. Among many potential candidates, MSP -3beta gene is promising target. This study was designed and carried out to determine the variation of this gene as genetic marker in population of malarious areas of Iran. Blood sample of 85 P. vivax isolates from four southern and east-southern provinces of the country assessed for polymorphism of PvMSP-3beta gene by PCR/RFLP method. Based on the size of PCR product of the gene, 7 genetically different types of parasite has been distinguished. Two alleles were simultaneously visible in 19% of the cases. Results from PCR/RFLP analysis of PvMSP-3beta gene showed at least 15 allelic groups. Multiple infections have been found in 2.4% of the cases. PvMSP-3beta gene was highly diverse in P. vivax isolates of malarious areas of Iran, and can be a suitable marker for population genetic studies of P. vivax. More investigations on PvMSP-3beta genes are needed to reveal genetic structure of P. vivax in Iran

Ophthalmomyiasis caused by flesh fly [Diptera: Sarcophagidae] in a patient with eye malignancy in Iran

Here we describe a case of Ophthalmomyiasis in a male patient with basal cell carcinoma. During the operation several live and motile maggots were removed from the lesion. Preliminary examination on the larvae confirmed their affiliation to the genus Sarcophaga [Diptera: Sarcophagidae].This genus is widely distributed throughout the world and species are very difficult to identify. The authors made attempt to approach species identification by rearing larvae to the adult flesh flies, but due to shortage of adult male specimen, reliable diagnosis in the level of species was not obtained. Possible interaction between ocular myiasis and malignancy concerning the case has not been addressed in this paper

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

Effects of anti-mosquito salivary glands and deglycosylated midgut antibodies of anapheles stephensi on fecundity and longevity

With the aim of controlling malaria by reducing vector population, the effects of antibodies produced against salivary glands and deglycosylated midgut antigens of Anopheles stephensi mosquitoes on fecundity and longevity of the same species were tested. Three deglycosylated preparations of midgut and two preparations of salivary glands were produced, conjugated with aluminum hydroxide gel, and subcutaneously injected to shoulders of TO [Turner Out-bred] mice. After 4 immunizations and assurance of enough antibody production against utilized antigenic suspensions, effects of blood feeding on immunized and control mice were assayed. Insoluble preparation of midgut showed the strongest effect with 23.5% reduction in egg laying, and increasing death rate of vectors in third day after feeding. No significant reduction in fecundity or survivorship was seen with other preparations. Anopheles midgut insoluble antigens are potential candidates for designing vaccines against malaria vectors and further investigations need to be done to find effective antigens and the best way of their use

In-vivo monitoring of the response of falciparum and vivax Plasmodia to chloroquine in Bandar-Abbas and Kahnoudj, South-East Iran, 1997-1999

The in-vivo 28 days extended test was applied for monitoring of the responses of P. falciparum and P. vivax to chloroquine in malaria patients referred to Malaria Research Laboratory in Bandar-Abbas Training and Health Research Center. The selected patients were treated with standard dose of chloroquine [25 mg/kg over 3 days]. Primaquine was also administered in a single dose [0.75 mg/kg in the third day] as gametocytocidal in falciparum and weekly [0.75 mg/kg/w for 8 weeks] as anti-relapse in vivax cases. From 76 falciparum malaria patients, 63 cases were followed up for 28 days; in 40 patients [63.5%] the parasites were resistant to chioroquine at RI and RII levels. In 13 patients that the asexual forms of P. falciparum were disappeared in their blood by the day 7 and they were not accessible more than one and two weeks, the response of the parasite was considered as either sensitive [S] or resistant at RI level [SRI]. The rate of the chioroquine-resistant cases of P. falciparum among Afghan refugees was higher [86.3%] than the rate in Iranian patients [51.2%]. Totally, there is no significantly difference of chloroquine-resistant rate and levels of P. falciparum in the studied areas in compare to the results of previous study. In 323 vivax malaria tested patients the mean of parasite clearance time [MPCT] was 2.91 days [ranged 1-5 days] and the parasite is still highly sensitive to chioroquine

Status of insecticide resistance in anopheles culicifacies [diptera: culicidae] in Ghasreghand district, Sistan and Baluchistan province, Iran

Anopheles culicifacies s.L plays an important role in transmission of malaria in Sistan and Baluchistan province, southeastern Iran. Adult susceptibility test on field-collected mosquitoes was conducted in Ghasreghand district. WHO diagnostic test procedures revealed that adult females were resistant to 0.4% dieldrin [mortality 64.5 +/- 3.13], tolerant to 0.1% propoxur [mortality 88.5 +/- 2.24] and susceptible to 4% DDT [mortality 98.75 +/- 0.8], 5% malathion [mortality 100%], 0.1% bendiocarb [mortality 98.86 +/- 0.7], 0.25% permethrin [mortality 98.4 +/- 0.1] and 0.1% lambdacyhalothrin [mortality 100%]. Malathion and lambdacyhalothrin had the highest efficacy against this species when they were exposed at the diagnostic dose for 1 hour followed by a 24 hour recovery period. Dieldrin, DDT and malathion had been used for malaria control as an indoor residual spraying. The implication of these findings in the control programme is discussed

Induction of drug resistance in Plasmodium falciparum: an intermittent drug exposure method

The production of experimentally induced drug resistance in the laboratory provides valuable opportunities for investigators to study the nature and genetics of drug resistance mechanisms to a given agent, patterns of cross resistance and the mode of action of drugs. At the beginning the continuous drug exposure was chosen as a standard procedure to produce drug' resistant strains of P. falciparum,.but later on some other methods were also applied. An intermittent drug exposure method as a novel procedure has been introduced in this study. Intermittent exposure of chloroquine resistant Kl and chloroquine sensitive T9.96 strains of P. falciparum to halofantrine culminated in a relatively rapid reduction in sensitivity to the drug. The response of halofantrifle - resistnat K1HF and T9.96 strains and parent parasites to halofantrifle, inefloquine, quinine and chloroquine was determined. The results indicated that the effectiveness of halofantrine to K1HF and T9.96HF strains decreased 9 and 3 folds respectively, compared to the parent parasites. Cross -resistance occurred among halofantrine. mefloquine and quinine. Halofantrine resistance was associated with enhanced chloroquine sensitivity in the strain derived from chloroquine - resistant K1 strain, hut not in the strain derived from chloroquine - sensitive T9.96 parasites