rate of plasmodium vivax infectivity within gloucose-6-phosphate dehydrogenase [G6PD] deficient individuals in hor-mozgan province, Iran

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

Evaluation of effectiveness of ethanolic extract of Artemisia aucheri, individually and in combination with chloroquine, on chloroquine - sensitive strain of Plasmodium berghei in sourian mice

Drug resistance in malaria parasites is extending in the world particularly in chemical synthesized drugs such as 4- aminoquinolines and aminoalcoholes. Employing herbal extracts is encouraged by WHO in the malarious areas. In this study, the effectiveness of ethanolic extract of Artemisia aucheri individually and in combination with chloroquine, has been considered against chloroquine - sensitive strain of Plasmodium berghei. At the first stage, ED50 of A. aucheri and chloroquine on P. berghei was calculated using in vivo test. Then based on the ED50s combination of A. aucheri and chloroquine with ratios of 0/100, 10/90, 20/80, 30/70, 40/60, 50/50, 60/40, 70/30, 80/20, 90/10 and 100/0 were tested against the parasite. For evaluating the adverse effect of A. aucheri on the mice, for two weeks 1000mg/kg of the extract was daily employed and the mice were followed up for fifty days. ED50s for chloroquine and A. aucheri were 1.6mg/kg and 1000mg/kg respectively. The outcome of two drugs combination on the mice showed antagonistic effects on the chloroquine - sensitive strain of parasite. Two weeks daily administration of A. aucheri had no toxic effect on the mice. A. aucheri individually can be effective in reducing the parasite while in combination with chloroquine loses its property

effect of otostegia persica in combination with chloroquine on chloroquine-sensitive and chloroquine-resistant strains of plasmodium berghei using in-vivo fixed ratios method

Malaria is one of the worldwide parasitic diseases which threaten the life of hundreds of millions of people at the malarious areas each year. The emergence of chloroquine-resistant strains of Plasmodium falciparum in most of the malarious areas has encountered the relevant countries with some difficulties about treating the acute cases of the disease particulary if the monotherapy regimen has been used. Because of many advantages for the combination therapy, the effectiveness of chloroquine [CQ] and Otostegia persica [OP], a medicinal plant in combination form, was tested against the chloroquine-sensitive and chloroquine-resistant strains of Plasmodium berghei in sourian mouse using in-vivo adapted fixed ratios method in this study. At the first step, ED[50]s [50% effective dose] of chloroquine and O. persica against both CQ-sensitive and CQ-resistant strains of P. berghei were calculated using in-vivo test in the mice. Ratios of 0, 10, 30, 50, 70, 90 and100% from each ED[50] were prepared and contrarily combined together to make the following fixed ratios of 0/100, 10/90, 30/70, 50/50, 70/30, 90/10, and 100/0 of CQ/OP and the parasites were exposed to the combined ratios. Determination of ED[50]s showed 1.1 mg/Kg and 2.4 mg/Kg of mouse body weight for chloroquine in CQ-sensitive and CQ-resistant strains respectively and 450 mg/Kg for O. persica in both strains. The results also showed that the combinations of "50% CQ + 50% OP", "30% CQ + 70% O.P" and "70% CQ + 30% OP" were more effective than other combinations against CQ-sensitive strain. The fixed ratio combinations of chloroquine and O. persica showed an additive in CQ-resistant strain. Toxicity consideration showed no toxic effect of the combinations on the mice. Otostegia persica potentiated the effectiveness of chloroquine against the chloroquine-sensitive strain of P. berghei but not on chloroquine-resistant P. berghei. Moreover, the greatly modified fixed ratios method in this study can be considered as useful methods for in-vivo combination tests in murine malaria parasites

A Recombinant Plasmodium vivax Apical Membrane Antigen-1 to Detect Human Infection in Iran

In Iran, Plasmodium vivax is responsible for more than 80% of the infected cases of malaria per year. Control interventions for vivax malaria in humans rely mainly on developed diagnostic methods. Recombinant P. vivax apical membrane antigen-1 (rPvAMA-1) has been reported to achieve designing rapid, sensitive, and specific molecular diagnosis. This study aimed to perform isolation and expression of a rPvAMA-1, derived from Iranian patients residing in an endemic area. Then, the diagnostic efficiency of the characterized Iranian PvAMA-1 was assessed using an indirect ELISA method. For this purpose, a partial region of AMA-1 gene was amplified, cloned, and expressed in pET32a plasmid. The recombinant His-tagged protein was purified and used to coat the ELISA plate. Antibody detection was assessed by indirect ELISA using rPvAMA-1. The validity of the ELISA method for detection of anti-P. vivax antibodies in the field was compared to light microscopy on 84 confirmed P. vivax patients and compared to 84 non-P. vivax infected individuals. The ELISA cut-off value was calculated as the mean+2SD of OD values of the people living in malaria endemic areas from a south part of Iran. We found a cut-off point of OD=0.311 that showed the best correlation between the sera confirmed with P. vivax infection and healthy control sera. A sensitivity of 81.0% and specificity of 84.5% were found at this cut off titer. A good degree of statistical agreement was found between ELISA using rPvAMA-1 and light microscopy (0.827) by Kappa analysis.

Genetic structure of Plasmodium vivax population assessed by sequence analysis of the merozoite surface protein 3 beta gene

The endemicity and transmission intensity levels of malaria are related to genetic diversity of the parasites. Merozoite surface protein 3beta [MSP3beta] is an important marker for assessing the polymorphic nature of Plasmodium vivax while it is also a vaccine candidate against the parasite. In this study we investigated the genetic structure of P. vivax population by sequence analysis of a polymorphic region of the P. vivax MSP3beta gene in isolates from Iran. Blood samples were collected from 100 patients with clinical symptoms. DNA was extracted and the target gene was amplified by polymerase chain reaction [PCR]. The sequences of 17 samples were used for sequence analysis using nucleotide Blast search and ClustalW multiple alignment. Phylogenetic tree was derived to describe the geographical branching and relationships. A large number of nucleotide insertions and deletions were observed in the sequences of polymorphic region of PvMSP3beta gene that were not specific in each biotype. Single nucleotide polymorphism [SNP] was found extensively in the sequences. The phylogenetic analysis did not show any significant geographical branching. The lack of any geographical branching and extensive polymorphism in MSP3beta gene of P. vivax isolates suggests that more investigations are needed to find a more suitable gene in order to develop a vaccine