Spironucleus muris and Eperythrozoon coccoides in Rodents from Northwestern Iran: Rare Infections

Background: Rodents perform a crucial role in dispersal of zoonosis causes globally. We aimed to investigation about infection levels of parasitic agents in rodents’ population in Meshkinshahr areas, northwest of Iran from Apr to Sep 2014. Methods: Two hundred four rodents were trapped and anaesthetized. A sample of blood was collected via cardi­opuncture from each one. Thin and thick blood smears were prepared and stained with Giemsa. All stained smear were examined under light microscopy with high magnification by two expert microscopists. Every suspected uni­cellular observed were measured microscopically and compared with key references to diagnose. Results: Captured rodents were identified as three genera including Meriones persicus, Mus musculus, Cricetulus migraturius. Protozoa identified in this study were included of Spironucleus muris and Eperythrozoon coccoides, these parasites were observed in blood smear of 0.98% of rodents. S. muris and E. coccoides were seen in M. mus­culus and C. migraturius, respectively. Conclusion: The present study increases awareness about Eperythrozoonosis in rodents and its potential transmis­sion to domestic animals and even to human in rural districts in Iran. Moreover, the attack of Spironucleus on the mucus of colon and its systemic risk was confirmed

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

Diagnosing Malaria Cases Referred to the Malaria Reference Laboratory in Tehran University of Medical Science, Iran

Background: 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. Methods: 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. Results: 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. Conclusion: 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

Determination of Asymptomatic Malaria among Afghani and Pakistani Immigrants and Native Population in South of Kerman Province, Iran

Background: 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. Methods: 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. Results: 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. Conclusion: 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.

Interaction between Chitosan and Chloroquine against Plas-modium berghei and P. falciparum using In-Vivo and In-Vitro Tests

Background: The use of antimalarial drugs with number of compounds in combination form may potentiate each other's activity. Methods: 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. Results: 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. Conclusion: 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

Endoparasites of Wild Rodents in Southeastern Iran

Background: This study was aimed to collect wild rodents for endoparasites determination in some parts of Sistan and Baluchistan Province, southeastern Iran nearby Pakistan and Afghanistan countries. Methods: A total of 100 wild rodents were captured alive with cage traps. Various samples were collected from blood and feces, also impression smear prepared from different organs. The samples were prepared by formalin-ether or stained with Giemsa, after that were examined under microscope. Results: All the caught rodents (47 Tatera indica, 44 Meriones hurriana, 5 Gerbilus nanus and 4 Meriones libycus) were studied for endoparasites emphasizing to their zoonotic aspects. Endoparasites including Spirurida, Hymenolepis diminuta, Hymenolepis nana feraterna, Trichuris trichiura, Skerjabino taenia, Trichostrongylus spp, Entamoeba muris, Chilomastix mesnili and Leishmania spp were parasitologically identified. Conclusion: Among 9 genera or species of the identified parasites at least 5 of them have zoonotic and public health importance

Antimalarial activity of Alcoholic Extract of Curcuma longa and Heracleum persicum on Cultivated Plasmodium falciparum 3D7 strain: Antimalarial Effect of C. longa and H. persicum

Introduction: Plasmodium falciparum causes the most fatal form of malaria in human. At present, the common treatments are not effective enough and the incidence of drug resistance is being increased in malarious areas. Therefore, presenting the novel methods for therapeutic purposes assumes significant importance.  Recent studies indicated that aqueous or alcoholic extracts of Curcuma longa and Heracleum persicum showed 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. Materials and Methods:  The alcoholic extracts of H. persicum and C. longa were prepared in 10-1, 10-3 and 10-5 mg/ml dilutions. These solutions were tested on P. falciparum with 10% parasitemia in RPMI 1640 medium with 10% hematocrit. Each of dilutions was examined in triplicate and the inhibitory effect of the solutions on parasites was measured via determining the average parasitemia and their schizont rate. Finally, the results were analyzed using SPSS software. Results: 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 significant effect on decreasing of schizont percentage in comparison with control group (P-value<0.05). Conclusions: the present study indicated that alcoholic extracts of C. longa and H. persicum possess acceptable antiplasmodial effects and could 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 notable in vitro antiplasmodial efficacy of C.longa and H.persicum, further studies with in vivo method is recommended

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

Background: 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.Methods: 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 and100/0 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 daysResults: 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.Conclusion: A. aucheri individually can be effective in reducing the parasite while in combination with chloroquine loses its property

DNA Sequence Polymorphism of the Lactate Dehydrogenase Genefrom Iranian Plasmodium vivax and Plasmodium falciparum Isolates

Background: 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. Methods: 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. Results: 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. Conclusion: 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

Considering the malaria is essential in a patient with suspected Ebola: case report

Background: 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. Case presentation: 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. Conclusion: 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