Surveillance and Vector Control of Lymphatic Filariasis in the Republic of Korea

ObjectivesUntil the early 2000s, lymphatic filariasis would commonly break out in the coastal areas in Korea. Through steady efforts combining investigation and treatment, filariasis was officially declared eradicated in 2008. This study surveyed the density of vector species of filariasis in past endemic areas, and inspected filariasis DNA from collected mosquitoes for protection against the reemergence of filariasis.MethodsBetween May and October 2009, mosquitoes were caught using the black night trap in past endemic coastal areas: Gyeongsangnam-do, Jeollanam-do, and Jeju-do. The collected mosquitoes were identified, and the extracted DNA from the collected vector mosquitoes was tested by polymerase chain reaction for Brugia malayi filariasis.ResultsOchletotatus togoi, Anophel es (Hyrcanus) group and Culex pipiens were most frequently caught in Jeollanam-do (Geomun Island, Bogil Island, Heuksan Island), Jeju-do (Namone-ri, Wimi-ri). and Gyeongsangnam-do (Maemul Island). DNA of B malayi was not found in Och Togoi and An (Hyrcanus) group as main vectors of filariasis.ConclusionLymphatic filariasis was not found in the vector mosquitoes collected in past endemic areas. However, considering that the proportion of vector species is quite high, there is a potential risk that filariasis could be reemerging through overseas travel or trade. Thus, there is a need to continuously monitor vector mosquitoes of lymphatic filariasis.     

Detection of Asymptomatic Carriers of Plasmodium vivax among Treated Patients by Nested PCR Method in Minab,Rudan and Bashagard,Iran

Background

Plasmodium vivax is the most widespread species of Plasmodium in humans and causing about 80 million clinical cases annually. This study was undertaken to detect P. vivax in asymptomatic treated vivax malaria patients to trace latent/sub-patent malaria infection.

Method

The venous blood of all detected cases with P. vivax in Bashagard, Minab and Roodan Districts in Hormozgan Province from 2009 to 2010 was examined by microscopic and nested PCR methods for presence of the parasite.

Results

In microscopic examination of peripheral blood smears, all samples were negative for the presence of the parasites. But, we detected two P. vivax related bands in the electrophoresis of the nested PCR products (120 bp).

Conclusion

Following up the malaria cases after treatment by a combination of methods, or new diagnostics such as RDTs can be included in the priorities of malaria elimination program in Iran.     

Plasmodium falciparum serine repeat antigen 5 (SE36) as a malaria vaccine candidate

A devastating disease spread by mosquitoes with high-efficiency, malaria imposes an enormous burden for which no licensed vaccine currently exists. Although the genome complexity of the parasite has made vaccine development tenuous, an effective malaria vaccine would be a valuable tool for control, elimination and eventual eradication. The Plasmodium serine repeat antigen 5 (SERA5) is an abundant asexual blood stage antigen that does not show any antigenic variation and exhibits limited polymorphism, making it a suitable vaccine candidate. Identified by comparing the IgG status of people in endemic areas with protective immunity and those with malaria symptoms, the vaccine potential of the N-terminal domain of Plasmodium falciparum SERA5 is also strongly supported by experimental data and immune responses both measured in vitro and in animal challenge models. The current understanding of SERA5 will be presented, particularly in relation to its path towards clinical development. The review highlights lessons learned and sorts out issues upon which further research efforts are needed.     

Increasing incidence of malaria in the Negro River basin, Brazilian Amazon

Malaria in Brazil is virtually restricted to the Amazon Region, where it has a heterogeneous geographic distribution. We reviewed secondary data in order to describe the regional and temporal distribution of 8018 malaria cases seen between 2003 and 2007 in Santa Isabel do Rio Negro, a municipality in the northwest Brazilian Amazon. A significant rise in malaria incidence, mainly in the Yanomami Indian reservation, was observed during this time. Anopheline breeding sites were also mapped and entomological data were obtained through the capture of larval and adult mosquitoes. Thirty-three potential breeding sites were identified in the urban and periurban areas, 28 of which were positive for anopheline larvae. Anopheles darlingi specimens were captured in both intra- and peridomicile locations in the urban areas. Demographic data were also assessed via a sectional survey, revealing that the majority of dwellings were vulnerable to mosquitoes. This study suggests that urban and periurban areas of this municipality are highly susceptible to epidemic malaria, which is endemic in the Yanomami Indian reservation near the city. In addition, transmission can be perpetuated autochthonously in the urban area, drawing attention to the continuous need for preventative measures such as controlling adult and aquatic stages of mosquitoes and improving housing.     

The Rate of Plasmodium vivax Infectivity within Gloucose-6-Phosphate Dehydrogenase (G6PD) Deficient Individuals in Hormozgan Province,Iran

Background

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.

Methods

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.

Results

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).

Conclusion

vivax malaria positive individuals with G6PD deficiency showed too mild symptoms of Malaria or even asymptomatic.     

Malaria Epidemiology in Mersin Province,Turkey from 2002 to 2011

Background

Malaria is an infectious disease caused by Plasmodium spp. with high morbidity and mortality in human in tropical and subtropical regions. In recent years, number of malaria cases has been significantly reduced because of fight with the disease in Turkey. This study intended to investigate the malaria epidemiology in Mersin Province from 2002 to 2011 using data from the provincial Public Health Directorate.

Methods

Over ten years, 303573 blood samples were taken from the people by active and passive surveillance methods and blood smears were prepared. Smears were stained with Giemsa and examined under the microscope.

Results

Totally, 73 people including 44 male and 29 female were positive in terms of Plasmodium spp. It was determined that P. vivax observed in 67 cases while P. falciparum in 6 cases. Cases were mainly observed in 15 to 44 years old range, showed an increase between June-September periods and a significant decrease after 2006. Out of the 73 malaria cases, 54 cases were from Mersin Province and 13 cases were imported from another province of Turkey. Six cases were transmitted from abroad.

Conclusion

These results provide information about malaria epidemiology in an endemic area in Turkey and contribute its prevention in Mersin Province.     

Unusual transmission of Plasmodium falciparum, Bordeaux, France, 2009

Plasmodium falciparum malaria is usually transmitted by mosquitoes. We report 2 cases in France transmitted by other modes: occupational blood exposure and blood transfusion. Even where malaria is not endemic, it should be considered as a cause of unexplained acute fever.     

Ecological transition from natural forest to tea plantations: effect on the dynamics of malaria vectors in the highlands of Cameroon

From October 2002 to September 2003, an entomological survey was carried out in a rural forested fringed village in the highlands of Mount Cameroon region to determine the temporal dynamics of the anopheline population and the intensity of malaria transmission. A total of 2387 Anopheles spp. were collected, with A. funestus predominating (59.9%), followed by A. hancocki (24.4%) and A. gambiae s.l. (15.7%). Considerable differences were observed in the nocturnal biting cycles of parous mosquitoes, with peak activity in the latter part of the night. PCR revealed that all specimens of the A. funestus group were A. funestus s.s. and all specimens from the A. gambiae complex were A. gambiae s.s. of the S molecular form. Plasmodium falciparum sporozoite rates of 17.3% and 8.5% were recorded for A. funestus and A. hancocki, respectively, with an anthropophilic rate of 96.3%. A strong positive correlation (r = 0.996) was found between the human-biting rate and the entomological inoculation rate (EIR). Malaria transmission was very high and perennial, with an estimated annual EIR of 460.1 infective bites per person per year. These results confirm that in high agricultural activity areas, A. funestus can be by far the major malaria vector responsible for malaria transmission.     

Immunization with genetically attenuated P. falciparum parasites induces long-lived antibodies that efficiently block hepatocyte invasion by sporozoites

Whole-parasite malaria vaccines have shown promise in clinical trials. We recently reported the first human trial of a malaria vaccine based on Plasmodium falciparum genetically attenuated parasites (PfGAP). Herein we report for the first time that PfGAP induces prolonged functional humoral responses in humans. Six volunteers were exposed to 5 bites of PfGAP-infected mosquitoes followed by approximately 200 bites. Plasma collected from all volunteers 3 months after the last exposure efficiently inhibits invasion of hepatocytes by P. falciparum sporozoites. The level of inhibition observed is comparable to that attained using plasma collected after 4–5 intravenously administrations of high numbers of irradiated sporozoites, validating the potential of PfGAP malaria vaccines. Our data highlight the role of antibody responses in pre-erythrocytic stages of human malaria, and suggests that to be protective, malaria vaccines might need to elicit long-lasting functional antibodies in addition to cellular responses.     

In vivo Susceptibility of Plasmodium vivax to Chloroquine in Southeastern Iran

Background

Plasmodium vivax is the predominant species causes of malaria with about 90% total annual reported malaria in Iran. This study conducted to determine the susceptibility of Plasmodium vivax isolates to chloroquine in Sistan and Balochistan Province, southeastern Iran.

Methods

A total 270 subjects with symptomatic malaria and confirmed P. vivax infection completed the designed 28-day in vivo study. The thick and thin film blood smears were screened for malaria parasites by microscopy. The nested PCR was applied using the Plasmodium 18 subunit ribosomal ribonucleic (Ssr RNA) genes for detecting mixed infections and diagnosis of parasites in the samples with low parasite on days 0, 5, 6, 7, and 28.

Results

P. vivax was cleared in 15%, 50%, 95%, and 100% of patients on days 1, 2, 3, 4 respectively by microscopy assessment. Six patients were exhibited specific P. vivax band in nested PCR on day 5. No recurrence was observed on days 7, 14 and 28. Mean (±standard deviation) parasite clearance time was 2.41 (±0.8) days.

Conclusion

P. vivax is still susceptible to chloroquine in Southeatern Iran. This finding is compatible with results of neighboring countries Pakistan and Afghanistan.     

Sequence Diversity of pfmdr1 and Sequence Conserve of pldh in Plasmodium falciparum from Indonesia: Its implications on Designing a Novel Antimalarial Drug with Less Prone to Resistance

Background

pfmdr1 and its variants are molecular marker which are responsible for antibiotics resistance in Plasmodium falciparum, a parasitic carrier for malaria disease. A novel strategy to treat malaria disease is by disrupting parasite lactate dehydrogenase (pLDH), a crucial enzyme for Plasmodium survival during their erythrocytic stages. This research was aimed to investigate and characterize the pfmdr1 and pldh genes of P. falciparum isolated from Nusa Tenggara Indonesia.

Methods

Genomic DNA of P.falciparum was isolated from malaria patients in Nusa Tenggara Indonesia. pfmdr1 was amplified using nested PCR and genotyped using Restriction Fragment Length Polymorphism (RFLP). pldh was amplified, sequenced, and analyzed using NCBI public domain databases and alignment using Clustal W ver. 1.83.

Results

Genotyping of the pfmdr1 revealed that sequence diversity was extremely high among isolates. However, a sequence analysis of pldh indicated that open reading frame of 316 amino acids of the gene showing 100% homology to the P. falciparum 3D7 reference pldh (GeneBank: {"type":"entrez-nucleotide","attrs":{"text":"XM_001349953.1","term_id":"124513265","term_text":"XM_001349953.1"}}XM_001349953.1).

Conclusion

This is the first report which confirms the heterologous of pfmdr1 and the homologous sequences of P.falciparum pldh isolated from Nusa Tenggara Islands of Indonesia, indicating that the chloroquine could not be used effectively as antimalarial target in the region and the pLDH-targeted antimalarial compound would have higher chance to be successful than using chloroquine for curbing malaria worldwide.     

Prediction Forecast for Culex tritaeniorhynchus Populations in Korea

Objectives

Japanese encephalitis is considered as a secondary legal infectious disease in Korea and is transmitted by mosquitoes in the summer season. The purpose of this study was to predict the ratio of Culex tritaeniorhynchus to all the species of mosquitoes present in the study regions.

Methods

From 1999 to 2012, black light traps were installed in 10 regions in Korea (Busan, Gyeonggi, Gangwon, Chungbuk, Chungnam, Jeonbuk, Jeonnam, Gyeongbuk, Gyeongnam, and Jeju) to capture mosquitoes for identification and classification under a dissecting microscope. The number of mosquitoes captured/week was used to calculate its daily occurrence (mosquitoes/trap/night). To predict the characteristics of the mosquito population, an autoregressive model of order p (AR(p)) was used to execute the out-of-sample prediction and the in-sample estimation after presumption.

Results

Compared with the out-of-sample method, the sample-weighted regression method''s case was relatively superior for prediction, and this method predicted a decrease in the frequency of Cx. tritaeniorhynchus for 2013. However, the actual frequency of this species showed an increase in frequency. By contrast, the frequency rate of all the mosquitoes including Cx. tritaeniorhynchus gradually decreased.

Conclusion

The number of patients with Japanese encephalitis has been strongly associated with the occurrence and density of vector mosquitoes, and the importance of this infectious disease has been highlighted since 2010. The 2013 prediction indicated an increase after an initial decrease, although the ratio of the two mosquito species decreased. The increase in vector density may be due to changes in temperature and the environment. Thus, continuous prevalence prediction is warranted.     

Assessment of microbial larvicide spraying with Bacillus thuringiensis israelensis, for the prevention of malaria

Objectives

The aim of this study was to assess the contribution of microbial larvicide spraying, Bacillus thuringiensis israelensis, as prevention strategy against malaria.

Methods

An experimental study consisted in spraying B. thuringiensis israelensis in a district during 1 year has been conducted. Another district (control) was not sprayed. Eight hundred and two children were evaluated, thick drop and swab examination was performed for those presenting with fever. The larval density was calculated in their habitats as well as larvicide remanence. Capture of mosquitoes with human bait allowed determining human exposure to bites at night, and identifying anopheles after dissection.

Results

The incidence of pediatric malaria was 13.8% in the sprayed district and 31.4% in the control district. The parasitic load ranged from 2000 to 42,000 parasites/μL in the sprayed district and 2000 to 576,000 parasites/μL in the control district. Plasmodium falciparum was the most frequent (97.8%) plasmodial species. In the control district, at least 20 larvae by liter of water were counted; anopheles larvae were found in 11 larval habitats out of 15 (73.33%). The human exposure to anopheles bites at night was 14.25 in the sprayed district and 33.13 in the control district. The remanence of B. thuringiensis israelensis was estimated at 9 days in the sprayed district.

Conclusion

The larvicide B. thuringiensis israelensis may be used in vector control strategy for the prevention of malaria.     

Malaria in Mazandaran,Northern Iran: Passive Case Finding During 1997-2012

Background

Malaria is one of the most important parasitic diseases in tropical and temperate regions. The aim of this study was to determine the trend of malaria in Mazandaran Province, northern Iran during 1997-2012.

Methods

This retrospective study was conducted from 1997 to 2012. The population''s study was individuals who registered at health centers of Mazandaran Province. Peripheral blood smear were prepared for each case, stained with Giemsa and examined by light microscope. In addition to demographic data, other parameters including Slide Positive Rate (SPR), Annual Parasite Incidence (API) and Annual Blood Examination Rate (ABER) were analyzed.

Results

In total, 844 cases of malaria were reported. Plasmodium vivax was predominant species with 821 cases (97.4%). The number of malaria cases increased from 1997 to 2005 and then decreased to 3 cases in 2011. Some cities had not reported any cases during last three years. The highest infection rate, 163(20.07%), was seen in 2001-02. The SPR had the highest value (0.54%) in 2004-05. The maximum API and ABER were observed in 2001-02 and 1997-98. 641(75.9%) of cases were imported from hyperendemic areas such as Afghanistan and South-eastern Iran and 94 (11.1%) malaria patients were recorded as introduced cases. The highest infection rate of malaria (21.3%) was seen in Babolsar.

Conclusion

Extensive malaria control should be continued to Mazandaran to become malaria-free region and in prevention of re-introduction stage.     

In Vivo Antimalarial Effects of Iranian Flora Artemisia khorassanica against Plasmodium berghei and Pharmacochemistry of its Natural Components

Background

The aim of this study was to evaluate the antimalarial effects of Iranian flora Artemisia khorassanica against Plasmodium berghei in vivo and pharmacochemistry of its natural components.

Methods

The aerial parts of Iranian flora A. khorasanica were collected at flowering stage from Khorassan Province, northeastern Iran in 2008. They were air-dried at room temperature; powder was macerated in methanol and the extract defatted in refrigerator, filtered, diluted with water, then eluted with n-hexane and finally non-polar components were identified through Gas Chromatography and Mass Spectroscopy (GC-MS). Toxicity of herbal extracts was assessed on naïve NMRI mice, and its anti-malarial efficacy was investigated on infected Plasmodium berghei animals. This is the first application on A. khorssanica extract for treatment of murine malaria. The significance of differences was determined by Analysis of Variances (ANOVA) and Student''s t-test using Graph Pad Prism Software.

Results

The herbal extract was successfully tested in vivo for its anti-plasmodial activity through artemisin composition, which is widely used as a standard malaria treatment.

Conclusion

Although, this study confirmed less anti-malarial effects of A. khorssanica against murine malaria in vivo, however there are some evidences on reducing pathophysiology by this medication. In complementary assay, major components were detected by GC-MS analysis in herbal extract including chrysanthenone (7.8%), palmitic acid (7.4%) and cis-thujone (5.8%). The most retention indices of the component are given as n-eicosane, palmitic acid and n-octadecane.     

Longitudinal and Cross-Sectional Genetic Diversity in the Korean Peninsula Based on the P vivax Merozoite Surface Protein Gene

Objectives

Vivax malaria has reemerged and become endemic in Korea. Our study aimed to analyze by both longitudinal and cross-sectional genetic diversity of this malaria based on the P vivax Merozoite Surface Protein (PvMSP) gene parasites recently found in the Korean peninsula.

Methods

PvMSP-1 gene sequence analysis from P vivax isolates (n = 835) during the 1996-2010 period were longitudinally analyzed and the isolates from the Korean peninsula through South Korea, the demilitarized zone and North Korea collected in 2008-2010 were enrolled in an overall analysis of MSP-1 gene diversity.

Results

New recombinant subtypes and severe multiple-cloneinfection rates were observed in recent vivax parasites. Regional variation was also observed in the study sites.

Conclusion

This study revealed the great complexity of genetic variation and rapid dissemination of genes in P vivax. It also showed interesting patterns of diversity depending, on the region in the Korean Peninsula. Understanding the parasiteninsula. Under genetic variation may help to analyze trends and assess the extent of endemic malaria in Korea.     

Noninvasive Test for Tuberculosis Detection among Primates

Traditional testing methods have limited epidemiologic studies of tuberculosis among free-living primates. PCR amplification of insertion element IS6110 of Mycobacterium tuberculosis from fecal samples was evaluated as a noninvasive screening test for tuberculosis in primates. Active tuberculosis was detected among inoculated macaques and naturally exposed chimpanzees, demonstrating the utility of this test.     

Report of a Consultation on the Optimization of Clinical Challenge Trials for Evaluation of Candidate Blood Stage Malaria Vaccines, 18–19 March 2009, Bethesda,MD, USA

Development and optimization of first generation malaria vaccine candidates has been facilitated by the existence of a well-established Plasmodium falciparum clinical challenge model in which infectious sporozoites are administered to human subjects via mosquito bite. While ideal for testing pre-erythrocytic stage vaccines, some researchers believe that the sporozoite challenge model is less appropriate for testing blood stage vaccines. Here we report a consultation, co-sponsored by PATH MVI, USAID, EMVI and WHO, where scientists from all institutions globally that have conducted such clinical challenges in recent years and representatives from regulatory agencies and funding agencies met to discuss clinical malaria challenge models. Participants discussed strengthening and harmonizing the sporozoite challenge model and considered the pros and cons of further developing a blood stage challenge possibly better suited for evaluating the efficacy of blood stage vaccines. This report summarizes major findings and recommendations, including an update on the Plasmodium vivax clinical challenge model, the prospects for performing experimental challenge trials in malaria endemic countries and an update on clinical safety data. While the focus of the meeting was on the optimization of clinical challenge models for evaluation of blood stage candidate malaria vaccines, many of the considerations are relevant for the application of challenge trials to other purposes.     

Differences in automated depolarization patterns of Plasmodium faiciparum and P. vivax malaria infections defined by the Cell-Dyn® CD4000 haematology analyser

Of 1014 samples submitted for full blood count analysis and malaria screening, 854 were designated malaria-negative by blood film microscopy, 79 were unequivocally identified as Plasmodium vivax and 81 as P. falciparum. All samples were additionally analysed with the Abbott Cell-Dyn® CD4000 haematology instrument, and leucocyte differential plots of 90° polarized vs. 90° depolarized (NEU-EOS plot) and 90° depolarized vs. 0° light (EOS I plot) scatter were specifically examined for abnormal depolarization patterns. Depolarization pattern types were correlated with microscopy (species) results, and these correlations were consolidated by polymerase chain reaction analysis. All 854 microscopically-designated malaria-negative samples showed a type 1 (normal) CD4000 depolarization pattern. Abnormal pattern types 2, 3a and 3b were entirely restricted to one of the two malaria categories. Plasmodium falciparum malaria showed two CD4000 pattern types only; a ‘normal’ type 1 pattern was seen in 36/75 (48%) cases and the remaining 39 cases were all abnormal pattern type 3a. In contrast, most (79/85) P. vivax malaria cases showed a distinctive clustered EOS I population (types 2 and 3b patterns) that was not seen with P. falciparum. Automated depolarization analysis provides an effective means of detecting malaria-associated haemozoin, and the patterns of intracellular haemozoin further appear to provide species differentiation between P. falciparum and P. vivax.     

Wash resistance and residual efficacy of long-lasting polyester netting coated with α-cypermethrin (Interceptor) against malaria-transmitting mosquitoes in Assam, northeast India

Malaria is endemic in Assam, northeast India, with low-to-moderate transmission of the causative parasites, mostly by Anopheles minimus. Plasmodium falciparum is the predominant parasite (>60%), with remaining cases being due to P. vivax. As an alternative intervention for malaria control, long-lasting insecticidal nets [Interceptor coated with α-cypermethrin 10% suspension concentrate (SC), 0.667% w/w, 0.2 g/m²] underwent field evaluation for laboratory wash resistance and residual efficacy in field conditions against malaria-transmitting mosquitoes. Based on entomological observations, the Interceptor net intervention was the most effective, corresponding to the lowest mosquito vector density in experimental villages. There was virtual disappearance of A. minimus in Interceptor net villages in contrast to the untreated net intervention and the no-net control. Contact cone bioassay tests revealed 100% mortality in the A.minimus group of mosquito species in the community using the Interceptor net, which was consistent during the follow-up monitoring period (October 2006 to April 2007) in field conditions. Similar levels of mortality were observed in laboratory-washed nets compared with unwashed nets, and wash resistance was consistent even after the 20th serial wash at fortnightly intervals. Community compliance and acceptance of the Interceptor net was high, with decreased nuisance due to biting mosquitoes and other household insect pests being reported.