afsaneh motevalli haghi

Iran is currently striving to eliminate malaria, making it essential to improve the accurate and rapid diagnosis of suspected cases.

This study aimed to evaluate the effectiveness of the multiplex/semi-nested polymerase chain reaction (PCR) method, using specific primers, for the precise diagnosis of human malaria species.

Seventy-two blood samples from patients and suspected malaria cases were selected and stored at -80°C in the National Malaria Laboratory. DNA extraction was performed to obtain the genetic material for further analysis. Multiplex/semi-nested PCR was conducted, and the results were compared with microscopic examination.

Out of 72 samples, 36 were positive by microscopic analysis, which included: 16 Plasmodium falciparum , 16 P. vivax , 1 P. ovale , 1 P. malariae , 1 mixed P. falciparum - P. vivax , and 1 mixed P. falciparum - P. malariae . Thirty-three cases were diagnosed through molecular analysis: 16 P. falciparum , 13 P. vivax , 1 P. ovale , 1 P. malariae , and 2 mixed P. falciparum - P. vivax .

Issues, such as false positives, underreporting of mixed infections, and mismatched species identified by microscopic methods need to be addressed and improved to ensure accurate diagnoses.

Resistance to artemisinin has threatened major achievements in malaria control, more investigations is needed about resistant strains and related genes. We aimed to induce resistance to artesunate in the Plasmodium falciparum 3D7 strain using intermittent exposure method and comparing P.fk13 gene sequence between susceptible and resistance strains.

P. falciparum 3D7 strain was cultured according to Trager & Jensen method with some modifications. Serial concentrations between 10-2 mol/l, to 10-7mol/l were prepared, then P. falciparum 3D7 was exposed to each of the dilution to determine IC50 and lethal dose. Sensitivity reduction process was started from the concentration of 10-7mol/l and ended at 10-2mol/l. Exposed parasites were collected after at least 27 days after cultivation in each drug concentration. DNA extraction, PCR and sequencing process were performed to investigate any possible mutations in the P.fk13 gene sequence.

Effectiveness of 10-2mol/l concentration of artemisinin was found as a lethal dose. IC50 value was equal to 5˟10-4 mol/l. The resistant strain was provided in the lab, sequenced and registered in the gene bank as P.f Art -2, (accession number MH796123. 1). Alignment of this registered sample showed no mutation in P.f kelch13 gene in comparison with standard strain submitted in the GenBank.

Resistance to artesunate in malaria parasite may occur but with no mutation in the P.f kelch13 gene. Therefore, whole genome sequencing should be applied to determine mutations in resistant strains.

Infections by Plasmodium falciparum, are becoming increasingly difficult to treat. Therefore, there is an urgent need for novel antimalarial agents’ discovery against infection. In present study, we described a 2’-O-Methyl gapmer phosphorothioate oligonucleotide antisense targeting translation initiation region of 3D7 strain RH5 gene.

The study was conducted in Pasteur Institute of Iran in 2020. ODNs effects were measured by microscopic examination and real time RT-PCR. For microscopy, microplates were charged with 2’-OMe ODNs at different dilutions. Unsynchronized parasites were added to a total of 0.4 ml (0.4% parasitemia, 5% red blood cells), and slides were prepared. Proportion of infected cells was measured by counting at least 500 red blood cells.

RH5 genes start codon regions selected as conserved region besed on alignment results. Gap-RH5-As which was complementary to sequence surrounding AUG RH5 start codon significantly reduced parasite growth (>90% at 50 nM) compared to sense sequence control (Gap-RH5-Se) (17%), (P<0.001). RH5 transcripts were dramatically reduced after exposed to ODNs at a concentration of 5-500 nM for 48 h.

Gemnosis delivery of a chimeric gapmer PS-ODN with 2’-OMe modifications at both sides had high antisense activity at low concentrations (10-100 nM) and shown a good efficiency to reach to target mRNA in human RBCs. Anti-parasite effect was correlated to reduction of target gene mRNA level. In addition, 2’-OMe ODNs free delivery is an effective way and does not need any carrier molecules or particles.

A variety of haemoprotozoa including Plasmodium, Haemoproteus and Leucocytozoon cause infections in birds and are transmitted by some known vectors. These parasites cause anemia, low appetite, weakness and ultimate ly death in birds. The present study was aimed to determine these parasites, in birds of Mazandaran and Golestan prov inces in Iran.

The project was performed on 340 live birds in 2016. The samples were collected from February to Septem ber 2016, from each bird, two thin and thick blood smears were prepared and the remaining blood about 1ml was kept in EDTA-containing tubes for molecular studies. The slides were stained with 10% Giemsa, then examined microscopical ly. About ten percent of the negative samples were considered for Polymerase Chain Reaction (PCR) technique, using specific primers to diagnose Plasmodium and Haemoproteus spp. Electrophoresis was done for PCR products and rele vant bands to the parasites were identified based on the size. The considered birds belonged to ducks, chickens, roosters, and pigeons.

From 340 microscopically examined blood samples 32 (9.5%) samples were positive. Twenty-five (7.35%) of them were infected with the genus Haemoproteus. Seven samples (14%) out of 50 microscopically negative samples were found as Haemoproteus or Plasmodium spp when PCR technique was employed.

This study revealed the existence of malaria parasites and other haemosporidia in birds in Iran. Employing molecular methods (PCR examination) could detect more infections.

Malaria parasites cause a tremendous burden of disease in both the tropics and subtropics areas. Growing of drugs resistance in parasites is one of the most threats to malaria control. The aim of study was to investigate the anti-malarial activity of nano-emodin isolated from Rhamnus cathartica on Plasmodium berghei in mice to evaluate parasites inhibition rate using in-vivo test.

The study was conducted in the School of Public Health, Tehran University of Medical Sciences, during 2020. Nano- emodin particles were prepared from Rhamnus cathartica, and confirmed by Zeta Potential Analyzer, DLS and electron microscopy techniques. Mice were infected with P. berghei and treated by emodin nanoparticles. Parasitemia was evaluated in each group in comparison with control group. Toxicity test was done using twice the highest concentration of emodin extract on a separate group of mice and ED50 was calculated.

Emodin extract was significantly effective in all concentrations on D4 (P<0.05). The most effective on parasitemia was observed in 400 mg/kg of Liquid Nano-emodin and solid (non-Nano) emodin. ED50 for emodin extract was determined 220 mg/kg. Toxicity test showed no toxic effect on the subjects.

The emodin extract is safe, lack of side effects. So, it can be used for more and longer period of time and in higher doses. Emodin extract, either in form of liquid and nanoparticle or in a solid form, has the same therapeutic effect on P. berghei in infected Balb/c mice.

Plasmodium falciparumcauses the most fatal form of malaria in humans. At present, the common treatments are not effective enough and the incidence of drug resistance is increasing in malarious areas. Therefore, presenting novel methods for therapeutic purposes assumes significant importance. Recent studies have indicated that aqueous or alcoholic extracts of Curcumalonga and Heracleum persicum show a broad spectrum of anti-microorganisms activity. In this (in vitro) study, the effects of C. longa and H. persicum extracts were assessed on P. falciparum since there has been limited clinical research into their effectiveness on Malaria.

The alcoholic extracts of H. persicum and C. longa were prepared in 10-1, 10-3, and 10-5mg/ml dilutions. These solutions were tested on P. falciparum with 10% parasitemia in RPMI 1640 medium with 10% hematocrit. Each of the dilutions was examined in triplicate and the inhibitory effect of the solutions on parasites was measured via determining the average parasitemia andtheir schizont rate. Finally, the results were analyzed using SPSS software.

The rate of parasitemia declined in three different dilutions of both H. persicum and C. longa. The mean of antiplasmodial inhibitory activity of herbs was 83.23±2.47% in H.persicum and 99.91±0.0% in C.longa. Moreover, all dilutions of both H.persicum and C.longa showed a significant effect on decreasing the percentage of schizont in comparison with the control group (P-value<0.05).

The present study indicated that alcoholic extracts of C. longa and H. persicum possess acceptable antiplasmodial effects and can be developed as valuable alternatives to ineffective antimalarial drugs. These results support the claims of recent studies that C. longa and H. persicum,have considerable antimicrobial activities.Considering thenotable in vitroantiplasmodial efficacy of C.longa and H.persicum, further studies with in vivo method are recommended.

The use of antimalarial drugs with number of compounds in combination form may potentiate each other's activity.

This study was conducted in the School of Public Health, Tehran University of Medical Sciences, Tehran, Iran in 2018. It was based on two methods including in vivo and in vitro tests with aim of considering interaction between chitosan and chloroquine against Plasmodium berghei and P. falciparum parasites using different ratios of the agents with ED50s and IC50s baselines.

Administrating 10 and 20 mg/kg (mouse body weight) of chitosan alone to the P. berghei –infected mice up to 4 successive days resulted in 37% and 45% inhibition of P. berghei respectively, while employing the compound with chloroquine in combination form with ratios of 90/10 and 70/30 (chloroquine/chitosan) had a considerable potentiation including 71.58% and 83.85% inhibition effectiveness against P. berghei. Moreover, 20 mg/L (CCM) concentration of chitosan alone could eliminate 69.55% of P. falciparum in culture medium while in combination with chloroquine in ratios of 90/10 (chloroquine/chitosan) had considerable potentiation including 79.14% inhibition effectiveness. Mean survival time of those mice received combination therapy in ratios of 90/10 and 70/30 (chloroquine/chitosan) was longer than those took up mono therapy of either chloroquine or chitosan based on their ED50s doses.

Interaction between chloroquine and chitosan showed considerable potentiation in combination form against either P. berghei or P. falciparum using in vivo and in vitro tests respectively. Meanwhile, interaction between the above mentioned agents resulted in a notable survival time for those P. berghei-infected mice treated with the combination.

Asymptomatic malaria, which usually exists in low parasitemia, acts as the Plasmodium species reser voirs contributing towards malaria transmission. This situation hinders malaria elimination programs in endemic areas, thus necessitating an active case detection with a high sensitive method and treatment of cases. This is why we used a High Resolution Melting (HRM) assay to monitor the trend of asymptomatic malaria in a malaria endemic area of Iran which is under elimination program.

The peripheral blood was sampled from 271 clinically approved non-febrile individuals from a malaria en demic zone of southeastern Iran for asymptomatic malaria prevalence detection by microscopy, Rapid Diagnostic Tests (RDTs) and HRM methods. The HRM assay was done based on the amplification of 18S SSU rRNA gene.

The HRM assay revealed infections from three individuals out of 271 (1.1% asymptomatic malaria prevalence) from the participants, two Iranian natives with Plasmodium vivax infection and one Pakistani immigrant with P. falcipa rum infection. Neither microscopy nor RDTs detected Plasmodium spp infections from the 271 non-febrile individuals. The nucleotide sequencing analysis of the positive controls used in this study showed a close homology with the refer ence gene bank sequences of P. falciparum 3D7 (CPO16995.1) and P. vivax Sal-1(UO3079.1).

This study revealed a low frequency of asymptomatic malaria trend within malaria endemic areas of southeastern Iran which are under intense elimination program and also the ability of HRM assay in detecting low Plasmodium spp parasitemia beyond the limits of microscopy and RDTs.

This study was designed to detect, if there are asymptomatic malaria infections amongst native and immigrant population from Afghanistan and Pakistan countries in Sistan & Baluchistan Province of Iran, where is under the national malaria elimination program.

This cross-sectional study was performed among native individuals and resident immigrants in the southeastern province of Sistan & Baluchistan from May 2016 to Jul 2017. A total of 271 individuals were considered in this cross- sectional study based on microscopical method, Rapid Diagnostic Tests (RDTs) and PCR techniques. Out of 271 native and immigrant participants 140 (52%) and 131 (48%) were male and female, respectively.

None of the prepared samples was diagnosed as malaria positive case when was considered via above mentioned three techniques.

Neither native nor immigrant individuals had asymptomatic malaria, hinting that national malaria elimination program is performed according to planned schedule in the studied areas

Among the human parasitic diseases, malaria is the main cause of morbidity and mortality. To prevent the high mortality and tracking malaria elimination efforts, a prompt and sensitive diagnosis is essential.  This study aimed to compare High-Resolution Melting (HRM) and microscopic methods to diagnose Plasmodium falciparum and P. vivax.

Eighty-one blood samples were collected from patients with clinical symptoms who were suspect to malaria in Chabahar district, southeastern Iran and also, from those who were referred to Malaria National Laboratory in the Tehran University of Medical Sciences, Tehran, Iran. Microscopic examination and HRM method were used to the diagnosis of Plasmodium parasites simultaneously.

Microscopic results revealed 45 positive cases (12 P. falciparum and 33 P. vivax) out of 81 collected samples while according to HRM analysis results 11 and 33 samples were identified as P. falciparum and P. vivax, respectively. HRM analysis also revealed 1 mixed infection of P. falciparum and P. malariae.

HRM analysis provides a promising mean for simultaneous detection and discrimination of the Plasmodium spp. especially in mixed infection cases.

Circumsporozoite protein (CSP) is one of the most important surface sporozoite antigens in malaria, recently considered as a candidate for vaccination. Considering the importance of CSP, this study was conducted to investigate the polymorphism and genetic diversity of Plasmodium vivax Circumsporozoite Protein (Pvcsp) in the southeastern region of Iran during 2015-2016.

To investigate polymorphism and genetic diversity, 20 blood samples were collected from patients with P. vivax, then DNA was extracted and amplified using partial sequence of CSP gene. Polymerase chain reaction (PCR) products were sequenced and compared to sequences from genomic databases using BLAST. Genetic evaluation and phylogenic analysis were performed using MEGA7 and DnaSP5 software’s on 38 sequences include 20 sequences of our study and 18 sequences of Gene Bank.

Eleven isolates were VK210 genotype and 9 isolates contained VK247. The result of variable segregation nucleotide site indicated that the differentiation of sequences in CSP were 25.67% in our 20 samples which are less than the 38 samples with a value of 26.67%. Comparing the ratio of dN/dS regions in the CSP gene indicates that the CSP varies more synonymously and amino acid has lower variation. Out of 38 samples, 35 unique haplotypes were identified based on 1042 nucleotide sequences in CSP, showing a variation percentage of 99.4%.

The Tajima D analyses showed that CSP gene in P. vivax had a positive number in the total analyzed sequences, which means that the P. vivax mutations are in order to select positive evolution.

Although Plasmodium vivax is usually known as benign malaria, some variations of the parasite can result in acute and sever infection. In this study we tried to determine some genetic variations in PvAMA-1 antigen among the samples were collected form southeastern Iran. About two ml blood samples were collected into EDTA pre-dosed tubes from 30 P. vivax–infected patients individually between 2011 and 2013. A Giemsa stained thick and thin blood film was prepared from each of the patients. A PCR-RFLP technique was employed using EcoR-1, Pvu-II and Hind3 restriction enzymes to determine the allelic variations of the antigen. A 1300bp gene corresponding to PvAMA-1 was selected for the amplification process. Among the total cases identified in this study 90% showed similar bounds when exposed to the restriction enzymes. Nine isolates (accession numbers: KF435081-KF435083 and JF682785-JF682790) were identified and registered in Gene bank. Identity among isolates was more than 96% in nucleotide level. Dendrogram clarified a close relationship among the clusters in spite of geographical distribution of the parasite. This study increased our data about prevalence and variation of PvAMA-1 alleles amongst P. vivax isolates in southeastern parts of Iran where besides native population bears considerable Afghan and Pakistani immigrants.

We evaluated the anti-malarial activity of Heracleum persicum individually and in combination with chloroquine.
This study was conducted at the School of Public Health, Tehran University of Medical Sciences, Tehran, Iran in 2015-2016. The Peter̛ s method was used for determining fifty percent effective dose (ED50) of the H. persicum extract and chloroquine individually against chloroquine sensitive P. berghei in small white mice. Six experimental groups for H. persicum and 6 groups for chloroquine and two control group (positive and negative) were considered for determination of ED50. Interaction between H. persicum and chloroquine also was evaluated based on fixed ratios method. Ratios of 0/100, 20/80, 40/60, 60/40, 80/20, 100/0 of ED50 of chloroquine and H. persicum respectively were tested against the parasite. Then inhibitory effects of two drugs were calculated and plotted in the relevant graphs.
Overall, 1500 mg/kg, and 1000 mg/kg concentrations of H. persicum against P. berghei resulted in ED50 and ED74 respectively. ED50 of chloroquine against the parasite was obtained as 1.4 mg/kg of mouse body weight. Moreover, combination of H. persicum and chloroquine showed a weak potentiation in ratios of 40/60 (chloroquine . persicum) with 64% inhibition, but not in other ratios.
Although H. persicum individually showed a reasonable antimalarial efficacy against chloroquine sensitive P. berghei, in combination with chloroquine it showed additive or antagonism result except in ratios of 40%CQ%HP.

Trichomonas muris is one of the most common protozoa diagnosed in rodents. The trichomonads are generally described as presenting only trophozoite form while pseudocyst is another morphological form of trichomonads identified among gastrointestinal and genitourinary trichomonads. We identified and described different shapes of T. muris pseudocysts and trophozoite in stool samples were collected from rodents including Merinos persicus, Mus musculus and Cricetulus migratorius.
In this cross-sectional study, stool samples from 204 trapped rodents were collected from Meshkin Shahr during Mar to Dec 2014. Samples were preserved in formalin 10% and PVA solution and transferred to Department of Medical Protozoology and Mycology, School of Public Health, Tehran University of Medical Sciences. Formalin-ether concentration method was used for the samples. The slides were stained with trichrome staining method and observed under light microscope.
The trophozoites were classified as T. muris based on size (18 to 24 µm), presence of three anterior flagella, recurrent flagellum, undulating membrane, and axostyle in direct examination and stained slides with trichrome staining method. Fifty-five out of 204 (27%) rodents were infected with T. muris in which 51(25%) samples pseudocysts form were observed. The spherical bodies of pseudocyst with almost 8 µm size, contained internalized flagella, an undulating membrane with recurrent flagellum, axostyle, and costa were seen. The pseudocysts were more prevalent than trophozoite form and pseudocysts were found with different shapes in this study.
T. muris pseudocysts were found in stool samples of caught rodents for the first time in northwestern Iran.

Astrodaucus persicus (Apiaceae) is one of the two species of this genus which grows in different parts of Iran. Roots of this plant were rich in benzodioxoles and used as food additive or salad in Iran and near countries. The aim of present study was evaluation of antimalarial and cytotoxic effects of different fractions of A. persicus fruits and roots extracts.
Ripe fruits and roots of A. persicuswere extracted and fractionated by hexane, chloroform, ethyl acetate and methanol, separately. Antimalarial activities of fractions were performed based on Plasmodium berghei suppressive test in mice model and percentage of parasitemia and suppression were determined for each sample. Cytotoxicity of fruits and roots fractions were investigated against human breast adenocarcinoma (MCF-7), colorectal carcinoma (SW480) and normal (L929) cell lines by MTT assay and IC50 of them were measured.
Hexane fraction of roots extract (RHE) and ethyl acetate fraction of fruits extract (FEA) of A. persicus demonstrated highest parasite inhibition (73.3 and 72.3%, respectively at 500 mg/kg/day) which were significantly different from negative control group (P According to the results, RHE and FEA fractions of A. persicus could be introduced as excellent choice for antimalarial drug discovery. In addition, cytotoxic activity of RHE was noticeable.

Rodents have important role as reservoirs of different parasites. The aim of this study was to determine helminth parasites of abundant rodents in Meshkin-Shahr, Ardabil Province northwest Iran.
From April 2014 to March 2015; 205 rodents including 118 Meriones persicus, 63 Mus musculus and 24 Cricetulus migratorius were collected, using live traps. All rodents were dissected and their different tissues examined for infectivity with helminth parasites.
Overall, 74.2% of rodents were infected with helminth parasites. The rate of infectivity in M. persicus, M. musculus and C. migratorius was 82.2%, 61.9%, 66.7%, respectively. In general, among all 205 rodents, the species and infection rates of helminthes were as follows: Nematoda: Trichuris sp. (46.8 %), Capillaria hepatica (18.1%), Syphacia frederici (14.2%), Aspicularis tetraptera (3.4%), Trichuris rhombomidis (2%), Heligmosomom sp. (2%), Streptopharagus kuntzi (0.5%), Spiruridae gen. sp. (0.5%); Cestoda: Hymenolepis nana fraterna (16.6%) Hymenolepis diminuta (7.3%) tetratiridium of Mesocestoides sp. (1%), Paranoplocephala sp. (0.5%), Cysticercus fasciolaris (0.5%), Taenia endothoracicus larva (0.5%), and Acanthocephala: Moniliformis moniliformis (18.5%).
Variable species of helminthes circulate in the rodents of the study area. Presence of several zoonotic species highlights the potential risk of infections for public health.

This study was proposed to monitor the situation of asymptomatic malaria among the native population and Afghani and Pakistani immigrants in Kahnooj and Ghale-Ganj districts from Kerman Province, Southeastern Iran.
A number of 180 and 120 individuals from Kahnooj and Ghale-Ganj respectively were registered and considered based on a cross-sectional surveillance method. From 300 registered cases, 200 individuals (66.7%) were selected among Afghani and Pakistani immigrants and the rest (33.3%) were native resident individuals. All samples were processed with employing microscopical examination, Rapid Diagnostic Tests (RDTs) and Semi- nested Multiplex PCR techniques.
None of the samples collected from native residents showed any malaria parasite, but among Afghani immigrants, one asymptomatic vivax malaria was detected in a 12 yr old girl with 280 parasites per microliter of blood. Moreover, one symptomatic vivax malaria was detected from a Pakistani immigrant with 47560 parasites per microliter of blood. All results obtained via microscopical method, confirmed by RDTs and PCR techniques.
To achieve the malaria elimination program different studies are needed that to be performed. Monitoring the asymptomatic malaria in all over the malaria endemic areas especially among the immigrant individuals is the most crucial necessity.

Parasite lactate dehydrogenase (pLDH) is extensively employed as malaria rapid diagnostic tests (RDTs). Moreover, it is a well-known drug target candidate. However, the genetic diversity of this gene might influence perfor­mance of RDT kits and its drug target candidacy. This study aimed to determine polymorphism of pLDH gene from Iranian isolates of P. vivax and P. falciparum. Genomic DNA was extracted from whole blood of microscopically confirmed P. vivax and P. falciparum infected patients. pLDH gene of P. falciparum and P. vivax was amplified using conventional PCR from 43 symptomatic malaria patients from Sistan and Baluchistan Province, Southeast Iran from 2012 to 2013. Sequence analysis of 15 P. vivax LDH showed fourteen had 100% identity with P. vivax Sal-1 and Belem strains. Two nucleotide substitutions were detected with only one resulted in amino acid change. Analysis of P. falciparum LDH sequences showed six of the seven sequences had 100% homology with P. falciparum 3D7 and Mzr-1. Moreover, PfLDH displayed three nucleotide changes that resulted in changing only one amino acid. PvLDH and PfLDH showed 75%-76% nucleotide and 90.4%-90.76% amino acid homology. pLDH gene from Iranian P. falciparum and P. vivax isolates displayed 98.8-100% homology with 1-3 nucleotide substitutions. This indicated this gene was relatively conserved. Additional studies can be done weather this genetic variation can influence the performance of pLDH based RDTs or not.

The number of malaria cases is declining worldwide; however, it remains as a serious health problem. Diagnosing unusual cases is the most im­portant issue to manage the problem. This study designed to describe the number of falciparum and vivax malaria infected patients referred to Malaria Reference Labora­tory in Tehran University of Medical Science from 2000 to 2012. A retrospective study was conducted based on the collected question­naires from each patient referred to the laboratory. Diagnosing results and demo­graphic information for positive cases were analyzed using SPSS software. Problem­atic cases were evaluated for any difficulties in diagnosis or in clinical signs. Scanning and molecular methods were performed whenever there was an atypical case referred to the laboratory. Some of the samples had various difficulties for diagnosing such as presence of fussed gametocytes and schizonts of Plasmodium falciparum in peripheral blood and CCHF like hemoragic disorders. Plasmodium vivax caused a large proportion of the cases (76.1%) in con­trast with P. falciparum that included smaller proportion (22.8%) and the rest (1.1) belonged to mixed infection. Most of the positive cases (69.6%) were belonged to Afghani people. Men (94.6%) showed more infection than women (5.4%), moreo­ver the most infection (44.5%) was seen at a range of 21-30 yr. In the case of existing atypical issues to diagnose, it is needed to per­form more precise microscopical examination beyond the current standard condi­tions. Sometimes molecular method is required to verify the exact agent of the dis­ease.

Malaria is still one of the major health problems in comparison with any other parasitic disease in Iran with considerable economic and mortality consequences. Sistan-and-Baluchestan, Hormozgan and Kerman are the most affected provinces in the country approximately 96% of the cases are reported from these three provinces. The aim of this study was to determine the frequency, distribution and rate of parasitaemia of human Plasmodium (P.) species in patients infected with malaria parasites in Kerman province. A total of 92,798 peripheral blood smears were collected from suspected malaria patients during the period 2009-10. Thin and thick blood smears were prepared according to the World Health Organization (WHO) standard procedure. Percentage of parasitaemia was determined based on the number of parasites in the positive slides. The Chi-square test was used for data analysis. A total of 571 samples were found to contain human Plasmodium species, including 523, 44, and 4 cases of P. vivax, P. falciparum and mixed infection, respectively. The results also showed that, as compared with the previous year, the total number of P. vivax cases decreased in 2010 by 33.96%. The highest level of parasitaemia was observed in one of the patients infected with P. falciparum, with 77240 parasites/µl of blood, and the lowest in a patient infected with P. vivax, with 48 parasites/µl of blood. There were no differences between the positive and negative cases as regards parameters such as nationality, habitat or gender (Chi-square, p<0.05). Furthermore, based on the Mann-Whitney test, there was no significant difference between the mean counts of P. falciparum and P. vivax (p-value = 0.464). Considering that Iran is in the elimination stage of malaria, patient finding and rapid, timely diagnosis of the disease are very important, particularly cases coming from Pakistan and Afghanistan, helping sustainability of the elimination program.

Asymptomatic malaria is a great challenge in the control، elimination and eradication programs of the disease in the endemic areas. The infected individuals with asymptomatic malaria are not cured and are، consequently، a potential source for contamination of the mosquito vectors and spread of the disease in the area. Therefore، detection of asymptomatic infected people is very important as regards combating the disease. This study was conducted to determine the presence and prevalence of asymptomatic malaria in Jask district، Hormozgan Province، Iran during 2012-13، in the hope that the results will help in designing strategies to eliminate the disease in the area. A total of 200 persons under coverage of health centers in Jask district were selected randomly and enrolled in the study. From each subject a 5-ml blood sample was taken in 3 occasions (total number of samples = 600)، slides p repared and examined using microscopic and molecular (PCR) methods، as well as rapid diagnostic (RDT) tests. None of the 600 slides prepared microscopically showed any positive malaria case. Neither did any of those prepared by RDTs or Nested-PCR. The findings of this study indicate that implementation of the malaria control program has been successful in the area; therefore the malaria elimination program should continue.

One of the main reasons of hemorrhagic fevers is Ebola. The high rate of mortality and lack of definite treatment have been caused this infection to be a serious problem in the world. Ebola, especially in the early stages, when causes symptoms such as fever, anorexia and nausea, can be confused with malaria infection and conversely, severe malaria with Ebola. Plasmodium falciparum is an important cause of severe malaria that more than other types of plasmodium confused with Ebola. The patient is a 54-year-old man who had gone to Sudan about 8 months ago. The patient reported that fever, chills and headache had been started one week before traveling from Sudan to Iran and hematuria was added to his symptoms in third week of illness in Iran. He was referred to the emergency department with probable diagnosis of Ebola. Plasmodium falciparum gametocytes were revealed in his peripheral blood smear. Finally, he was treated with Coartem (artemether/lumefantrine) for malaria and after clinical improvement discharged to home with good condition. Ebola should be suspected in every patient with fever and a history of traveling to endemic areas. Considering the fact that in most areas where Ebola is endemic also malaria is common, lack of clinical suspicion to malaria causes that clinicians mistake malaria with Ebola. Necessary laboratory tests to rule out important differential diagnoses in patients with suspected Ebola virus contains: Peripheral blood smear for malarial parasite and blood culture and blood cell counts to investigate typhoid fever and other bacterial infections. Therefore, malaria should be considered as an important differential diagnosis in every patient suspected with Ebola.

One of the most important enzymatic disorders that interact with malaria is deficiency of G6PD (Gloucose-6-phosphate dehydrogenase). This enzyme protects red blood cells from hydrogen peroxide and other oxida­tive damages. Distribution of this enzyme deficiency usually accompa­nies with low level distribution of malaria disease in most malarious areas. So this hypothesis may be considered that the G6PD deficiency could be protec­tive against malaria. Totally 160 samples were taken from vivax malaria infected and non-infected individuals. Preparing blood smears and quantitative test for G6PD deficiency were employed for all of the samples. To ensure accuracy of the malaria in negative samples besides using microscopical examination, semi-nested multiplex PCR was also performed for the two groups. In microscopical examination 36 and 124 samples were vivax ma­laria positive and negative respectively. Out of 36 P.vivax positive cases 3 (8.3%) cases were detected to be G6PD deficient versus 30 (24.2%) cases out of 124 P. vivax negative cases. The results showed a significant differentiation between P. vivax positive and P. vivax negative cases in the rate of G6PD defi­ciency (3/36 in positive cases versus 30/124 in negative cases) (P<0.05). vivax malaria positive individuals with G6PD deficiency showed too mild symptoms of Malaria or even asymptomatic.

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 4th & 7th 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.