Assessment and Management of Mercury Leaching from a Riverbank.

The South River located in the city of Waynesboro, Virginia, contains mercury (Hg) contamination due to historical releases from an industrial facility operating between 1929 and 1950. In 2015, two sampling events were conducted in two of the contaminated bank regions (Constitution Park and North Park) to evaluate non-particulate total mercury (THg) and methylmercury (MeHg) concentrations in bank interstitial waters during river base flows and during bank drainage after flooding events. Porewater THg and MeHg at the bank-water interface were measured using diffusive gradient in thin-film devices (DGTs). The results showed THg mercury concentrations during bank drainage were approximately a factor of 3 higher than during base flow conditions. To have a better understanding of the parameters that control Hg leaching, a series of laboratory experiments were designed using South River sediments. The field and laboratory assessment showed that drainage/inundation cycles can lead to high THg concentration leachate from contaminated sediment due to increased partitioning from solids under oxic bank conditions and mobilization by the drainage waters. The results also demonstrated that methyl mercury concentrations at the bank-water interface are highest under base flow when conditions are more reduced due to the absence of oxic water exchange with the surface water. A remedial approach was implemented involving partial removal of surficial sediments and placement of biochar (to reduce non-particulate THg) and an armoring layer (to reduce erosion). DGT Measurements after bank stabilization showed THg decreased by a factor of ~200 and MeHg concentration by a factor of more than 20.

Disparate selection of mutations in the dihydrofolate reductase gene (dhfr) of Plasmodium ovale curtisi and P. o. wallikeri in Africa.

Plasmodium ovale curtisi and P. ovale wallikeri are both endemic in sub-Saharan Africa, the Middle East and Southeast Asia. Molecular surveillance data for drug resistance in P. ovale spp. is limited at present. We analysed polymorphisms in the podhfr, pocrt and pocytb genes of P. ovale spp. in 147 samples collected from travelers returning to China from Africa. Two podhfr mutations, S58R and S113N/T were detected in P. ovale curtisi with high/moderate frequencies of 52.17% and 17.39%, respectively. Evidence of positive selection (dN/dS = 2.41) was found for podhfr in P. ovale curtisi and decreased diversity (He) of microsatellite markers flanking the mutant alleles suggests that selective sweeps have occurred for both. Mutations E34G (1.50%) and L43V (1.50%) in pocrt of P. ovale curtisi, and E34G (3.70%), I102M (1.80%) and V111F (1.80%) of P. ovale wallikeri were found at low frequencies. Mutations R66K (6.20%), R75K (11.63%) and R95K (3.88%) of pocytb were found in both P. ovale curtisi and P. ovale wallikeri. These results suggest that the podhfr gene of P. ovale curtisi may be subject to drug selection in Africa, warranting further attention. We observed significant differences in the prevalence and distribution of podhfr mutations between the two P. ovale species, suggestive of fundamental biological differences between them.

Case-based malaria surveillance and response: implementation of 1-3-7 approach in Jiangsu Province, China.

Following initiation of China's National Malaria Elimination Action Plan (NMEAP) in 2010, China's 1-3-7 surveillance and response approach was developed and rolled out in China to facilitate the malaria control programme and accelerate the progress of malaria elimination. Innovative strategies and interventions have been developed and implemented in Jiangsu Province to facilitate case-based malaria surveillance and response. A total of 9879 malaria cases were reported in Jiangsu Province from 2001 to 2020. Since 2012, no indigenous malaria cases have been reported in Jiangsu Province. However, in recent years, there has been a substantial increase of imported cases from abroad. To continue improving the malaria surveillance and response system, Jiangsu Province has conducted population-based health education to improve the healthcare seeking behaviour of malaria patients, strengthened the capacity of health facilities to improve the performance of malaria diagnosis and treatment, and strengthened health workforce capacity to improve the implementation of 1-3-7 approach. Continually improving surveillance and response system can play a critical role in the early detection and rapid response of individual malaria cases and prevent the re-establishment of malaria.

Differential metabolome responses to deltamethrin between resistant and susceptible Anopheles sinensis.

Insecticide-based vector control measures play an important role in the prevention and control of insect-borne infectious diseases such as malaria; however, insecticide resistance has become a severe global problem for vector control. To date, the metabolic mechanism by which Anopheles sinensis, the most widely distributed malaria vector in China and Asia, detoxifies insecticides is not clear. In this study, the molecular metabolite changes in both the larval and adult stages of deltamethrin susceptible (DS) and deltamethrin-resistant (DR) An. sinensis mosquitoes were analysed by using liquid chromatography tandem mass spectrometry (LC-MS/MS) after exposure to deltamethrin. There were 127 differential metabolites in larval DR An. sinensis and 168 in adults. Five metabolites (glycerophosphocholine, deoxyguanosine, DL-methionine sulfoxide, D-myo-inositol-3-phosphate and N-acetyl-alpha-D-glucosamine1-phosphate) were downregulated in both DR larvae and adults, and one metabolite (aspartyl-glutamine) was upregulated, and the ratio of down- and up-regulation of these metabolites was 5:1. The differential metabolites between the DS and DR mosquitos were mainly classified into organic oxygen compounds, carboxylic acids and their derivatives, glycerophospholipids and purine nucleotides, and the common pathway enriched in both the larval and adult DR An. sinensis was glycerophospholipid metabolism. The findings of this study provide further mechanistic understanding of insecticide resistance in An. sinensis.

The complete mitochondrial genome of major malaria vector Anopheles anthropophagus (Diptera: Culicidae) in China.

Anopheles anthropophagus (Xu and Feng 1975) is the major vector of malaria in Eastern and Southern China. The species An. anthropophagus is considered a synonym of An. lesteri (Baisas & Hu, 1936), although they differ in several key biological characteristics. Here, we report the complete mitochondrial genome of An. anthropophagus for the first time. The mitogenome of An. anthropophagus is a typical circular, double-stranded molecule with a total length of 15,413 base pairs, and contains 13 protein-coding genes, 22 transfer RNA genes, two ribosomal RNA genes, and an AT-rich control region. A phylogenetic analysis of the complete mitogenomes of 16 species of Anopheles (Culicidae) revealed that An. anthropophagus is closely related to An. sinensis (Wiedemann 1828), in the family Culicidae. The An. anthropophagus mitogenome provides new data for further taxonomic and phylogenetic studies of the genus Anopheles.

Improving the surveillance and response system to achieve and maintain malaria elimination: a retrospective analysis in Jiangsu Province, China.

BACKGROUND: Following initiation of China's National Malaria Elimination Action Plan (NMEAP) in 2010, the '1-3-7' approach was developed and rolled out in China to facilitate the malaria elimination programme and accelerate malaria elimination. This study aims to summarize and condense these experiences through a retrospective analysis in Jiangsu Province, which could be adapted and applied in other malaria elimination settings worldwide. METHODS: A retrospective analysis of imported malaria cases into China identified through an improved surveillance and response system in Jiangsu Province was carried out for the period of 2001-2020. To improve the malaria surveillance and response system, Centers for Diseases Control and Prevention from the prefectures and counties in Jiangsu province conducted population-level health education to improve healthcare seeking behavior, strengthened capacity of health facilities to improve performance of malaria diagnosis and treatment, and raised the capacity of public health providers to improve implementation of the '1-3-7' approach. Categorical variables were carried out by Chi square tests with Fisher's exact correction. RESULTS: From 2001 to 2020, a total of 9,879 malaria cases were reported in Jiangsu Province. Since 2012, no indigenous malaria cases have been reported in Jiangsu Province. However, in recent years, there has been a substantial increase of imported falciparum malaria cases. Between 2012 and 2020, an estimated 61.57 million individuals have benefited from population-level health education in Jiangsu Province. For healthcare-seeking services among the 2,423 imported malaria cases, 687 (28.4%) and 1,104 (45.6%) cases visited hospitals on the first day and the second day from symptom onset, respectively. A total of 1,502 (61.9%) cases were diagnosed on the first day at medical facilities. Jiangsu Province achieved 100%, 99.4% and 98.3% completion rate in terms of case detection and notification (within one day), case investigation (within three days) and foci response and disposition (within seven days), respectively. The improved surveillance and response system in Jiangsu Province plays an important role in preventing the re-introduction of malaria and maintaining the malaria-free status. CONCLUSIONS: Jiangsu Province has maintained its malaria-free status since 2012. The continuous improvement of a surveillance and response system plays an important role in the early detection and rapid response of potential malaria-related outbreaks in Jiangsu, China, and has important lessons for other malaria eliminating settings. Remaining vigilant in the detection of imported malaria cases and maintaining an active surveillance and response system is critical to sustain the success of malaria elimination.

A PCR-Based Technique to Track the Geographic Origin of Plasmodium falciparum With 23-SNP Barcode Analysis.

Increased population movement has increased the risk of reintroducing parasites to elimination areas and also dispersing drug-resistant parasites to new regions. Therefore, reliable and repeatable methods to trace back to the source of imported infections are essential. The recently developed 23-single-nucleotide polymorphism (SNP) barcode from organellar genomes of mitochondrion (mt) and apicoplast (apico) provides a valuable tool to locate the geographic origin of Plasmodium falciparum. This study aims to explore the feasibility of using the 23-SNP barcode for tracking P. falciparum by polymerase chain reaction and sequencing, while providing geographical haplotypes of isolates that originated from Central Africa. Based on 23-SNP barcode analysis, SNPs were found at seven loci; 27 isolates were confirmed to have originated in West Africa, and this study also showed four isolates from Central Africa (Equatorial Guinea, 3; Republic of Congo, 1) that originated in East Africa. This study provides the sequence data from Central Africa and fills 23-SNP barcode data gaps of sample origins.

Cost-effectiveness analysis of malaria rapid diagnostic test in the elimination setting.

BACKGROUND: As more and more countries approaching the goal of malaria elimination, malaria rapid diagnostic tests (RDT) was recomendated to be a diagnostic strategy to achieve and maintain the statute of malaria free, as it's less requirments on equipment and experitise than microscopic examination. But there are very few economic evaluations to confirm whether RDT was cost-effective in the setting of malaria elimination. This research aimed to offer evidence for helping decision making on malaria diagnosis strategy. METHODS: A cost-effectiveness analysis was conducted to compare RDT with microscopy examination for malaria diagnosis, by using a decision tree model. There were three strategies of malaria diagnostic testing evaluated in the model, 1) microscopy, 2) RDT, 3) RDT followed by microscopy. The effect indicator was defined as the number of malaria cases treated appropriately. Based on the joint perspective of health sector and patient, costs data were collected from hospital information systems, key informant interviews, and patient surveys. Data collection was conducted in Jiangsu from September 2018 to January 2019. Epidemiological data were obtained from local malaria surveillance reports. A hypothetical cohort of 300 000 febrile patients were simulated to calculate the total cost and effect of each strategy. One-way, two-way, and probabilistic sensitivity analysis were performed to test the robustness of the result. RESULTS: The results showed that RDT strategy was the most effective (245 cases) but also the most costly (United States Dollar [USD] 4.47 million) compared to using microscopy alone (238 cases, USD 3.63 million), and RDT followed by microscopy (221 cases, USD 2.75 million). There was no strategy dominated. One-way sensitivity analysis reflected that the result was sensitive to the change in labor cost and two-way sensitivity analysis indicated that the result was not sensitive to the proportion of falciparum malaria. The result of Monte Carlo simulation showed that RDT strategy had higher effects and higher cost than other strategies with a high probability. CONCLUSIONS: Compared to microscopy and RDT followed by microscopy, RDT strategy had higher effects and higher cost in the setting of malaria elimination.

Confirmation of the absence of local transmission and geographic assignment of imported falciparum malaria cases to China using microsatellite panel.

BACKGROUND: Current methods to classify local and imported malaria infections depend primarily on patient travel history, which can have limited accuracy. Genotyping has been investigated as a complementary approach to track the spread of malaria and identify the origin of imported infections. METHODS: An extended panel of 26 microsatellites (16 new microsatellites) for Plasmodium falciparum was evaluated in 602 imported infections from 26 sub-Saharan African countries to the Jiangsu Province of People's Republic of China. The potential of the 26 microsatellite markers to assign imported parasites to their geographic origin was assessed using a Bayesian method with Markov Chain Monte Carlo (MCMC) as implemented in the program Smoothed and Continuous Assignments (SCAT) with a modification to incorporate haploid genotype data. RESULTS: The newly designed microsatellites were polymorphic and are not in linkage disequilibrium with the existing microsatellites, supporting previous findings of high rate of recombination in sub-Saharan Africa. Consistent with epidemiology inferred from patients' travel history, no evidence for local transmission was found; nearly all genetically related infections were identified in people who travelled to the same country near the same time. The smoothing assignment method assigned imported cases to their likely geographic origin with an accuracy (Angola: 59%; Nigeria: 51%; Equatorial Guinea: 40%) higher than would be achieved at random, reaching statistical significance for Angola and Equatorial Guinea. CONCLUSIONS: Genotyping using an extended microsatellite panel is valuable for malaria case classification and programme evaluation in an elimination setting. A Bayesian method for assigning geographic origin of mammals based on genetic data was adapted for malaria and showed potential for identification of the origin of imported infections.

Evaluating the implementation of rapid diagnostic tests in a malaria elimination setting.

BACKGROUND: It was recommended that malaria rapid diagnostic tests (RDTs) should be available in all epidemiological situations. But evidence was limited on the implementation of RDTs and its effectiveness in malaria elimination settings. This study examined the implementation of RDTs and how it affected the diagnosis of imported malaria patients in Jiangsu Province, China. METHODS: To scale up RDTs, this study developed an intervention package with four major elements covering the supply of RDT test, the training on RDTs, the monitoring and management of RDT use, and the advocacy of RDTs. By using a pretest-posttest control group design, we implemented the interventions in 4 cities in Jiangsu Province with the rest nine cities as controlled areas, from January 2017 to January 2018. Difference-in-Difference approach was used to evaluate the impact of the scale-up of RDTs on the identification of malaria cases. Three binary outcome measures were included to indicate delayed malaria diagnosis, malaria cases with confirmed malaria diagnosis at township-level institutions, and severe malaria cases, respectively. Linear probability regression was performed with time and group fixed effects and the interaction term between time and group. RESULTS: Intervention areas received sufficient RDT test supply, regular professional training programs, monthly tracking and management of RDT supply and use, and health education to targeted population. The implementation of interventions was associated with 10.8% (P = 0.021) fewer patients with delayed diagnosis. But intervention areas did not see a higher likelihood of having confirmed diagnosis from township-level institutions (coefficient = -0.038, P = 0.185) or reduced severe malaria cases (coef. = 0.040, P = 0.592). CONCLUSIONS: The comprehensive package of RDT implementation in this study is promising in scaling up RDT use and improving access to care among malaria patients, especially in malaria elimination settings.

Classification of induced malaria case in an elimination setting: investigation of transfusion-transmitted malaria cases.

BACKGROUND: Since the National Malaria Elimination Action Plan was launched in China in 2010, local malaria transmission has decreased rapidly. Zero indigenous cases were reported since 2017. However, after 2010, the proportion of imported cases in China increased from 45.7% in 2010 to 99.9% in 2016, and almost all provinces of China have reported imported cases in recent years. Prevention of the reintroduction of malaria into China is crucial for the maintenance of its malaria-free status. Hence, it is of utmost importance to correctly identify the source of malaria infections within the country. CASE INTRODUCTION AND RESPONSE: In 2016 and 2017, three laboratory-confirmed cases of malaria caused by Plasmodium falciparum were identified in patients with no previous travel history to endemic areas were reported in Jiangsu Province, China, where malaria due to P. falciparum was eliminated about 30 years ago. These were diagnosed after 41, 31 and 39 days of seeking treatment, respectively, and all of them had received blood transfusions. Further investigations indicated that two of the cases had received blood from foreign students (from Indonesia and Ghana), and the other had received blood from an individual who had worked in Equatorial Guinea. All three blood donors were traced, and found to be carrying asymptomatic P. falciparum infections by microscopic examination and PCR. Furthermore, five polymorphic microsatellite markers (C1M4, C4M62, C13M13, C14M17, and C13M63) were typed and used to link parasites from the donors with those of the transfusion-receiving patients. CONCLUSIONS: Three transfusion-transmitted malaria cases were identified in China, all of which were due to the transfusion of blood donated by individuals who had contracted malaria outside the country. These cases can provide a reference for those faced with similar challenges in malaria case identification and classification in other regions. In addition, a stricter screening policy including the use of appropriate detection methods for malaria parasites should be developed and adopted for blood donation in regions undergoing malaria elimination.

Malaria Elimination in China: Improving County-Level Malaria Personnel Knowledge of the 1-3-7 Strategy through Tabletop Exercises.

As China moves to the prevention of reestablishment of malaria, maintaining skills for malaria in county personnel on the "1-3-7" surveillance and response strategy is critical. China's "1-3-7" strategy defines targets used to guide and monitor malaria case reporting, investigation, and response, respectively: reporting of malaria cases within 1 day, their confirmation and investigation within 3 days, and the appropriate public health response to prevent further transmission within 7 days. Assessing the knowledge of local CDC malaria personnel on the "1-3-7" surveillance and response strategy is urgently needed. In June 2016, two different training modules (classroom-style teaching and tabletop exercises) were conducted for 125 CDC staff in Jiangsu Province, China, to determine the effectiveness of the two training modules on CDC staff knowledge and learning of the "1-3-7" strategy. The classroom-style training module just imparted the malaria knowledge to participants through teaching. Tabletop exercises were carried out through discussion-based scenarios using questions and answers on the "1-3-7" strategy. Questionnaires assessing knowledge improvement were designed and administered to personnel responsible for malaria surveillance and response activities, including at baseline and end line. Overall, knowledge of the "1-3-7" strategy for malaria elimination was 63.2% correct at baseline, 70.6% after implementing a classroom-style teaching module (χ2 = 11.20, P = 0.001), and 84.6% after the tabletop exercise module (χ2 = 48.82, P < 0.001). The knowledge of each component of the "1-3-7" strategy improved significantly after the tabletop exercise module. The total proportion of respondents with a high score (greater than or equal to 75%) was 82.7% in the classroom-style module and 95.2% in the tabletop exercise module. The proportion of respondents with a high score significantly increased after tabletop exercises in the stratified demographic groups of men who work at the county CDC level, have a bachelor's degree, hold a professional title as professor or assistant, are aged 31-50 years, and have attained 11-20 years of service with the CDC compared with the classroom-style module. Acceptability of the classroom-style module (78.2%) compared with tabletop exercises (94.4%) by the CDC malaria personnel increased significantly (χ2 = 11.96, P = 0.004). Feedback from participants on the modules suggest the tabletop exercises were an effective training method, which could maintain and improve the knowledge and capacity for malaria surveillance and response in basic CDC level personnel in China.

Assessment of false negative rates of lactate dehydrogenase-based malaria rapid diagnostic tests for Plasmodium ovale detection.

Currently, malaria rapid diagnostic tests (RDTs) are widely used for malaria diagnosis, but test performance and the factors that lead to failure of Plasmodium ovale detection are not well understood. In this study, three pLDH-based RDTs were evaluated using cases in China that originated in Africa. The sensitivity of Wondfo Pf/Pan, CareStart pLDH PAN and SD BIOLINE Pf/Pan in P. ovale detection was 70, 55 and 18%, respectively. CareStart was worse at detecting P. o. curtisi (36.5%) than at detecting P. o. wallikeri (75.0%), and SD could not detect P. o. curtisi. The overall detection ratio of all three RDTs decreased with parasite density and pLDH concentration. Wondfo, CareStart and SD detected only 75.0, 78.1 and 46.9% of the P. ovale cases, respectively, even when the parasitemia were higher than 5000 parasites/µL. Subspecies of P. ovale should be considered while to improve RDT quality for P. ovale diagnosis to achieve the goal of malaria elimination.

Plasmodium ovale curtisi and Plasmodium ovale wallikeri in Chinese travelers: Prevalence of novel genotypes of circumsporozoite protein in the African continent.

Imported malaria due to Plasmodium ovale curtisi and P. ovale wallikeri infections from African countries has increased recently (2011-2014) in Chinese travelers. We report novel genotypes, their prevalence and the predominant haplotypes of P. ovale curtisi and P. ovale wallikeri circumsporozoite protein (CSP) from 20 African countries in Chinese travelers. These genotypes should be considered while designing a CSP-based vaccine against P. ovale malaria.

Cost-effectiveness analysis of malaria rapid diagnostic tests: a systematic review.

BACKGROUND: Rapid diagnostic tests (RDT) can effectively manage malaria cases and reduce excess costs brought by misdiagnosis. However, few studies have evaluated the economic value of this technology. The purpose of this study is to systematically review the economic value of RDT in malaria diagnosis. MAIN TEXT: A detailed search strategy was developed to identify published economic evaluations that provide evidence regarding the cost-effectiveness of malaria RDT. Electronic databases including MEDLINE, EMBASE, Biosis Previews, Web of Science and Cochrane Library were searched from Jan 2007 to July 2018. Two researchers screened studies independently based on pre-specified inclusion and exclusion criteria. The Consolidated Health Economic Evaluation Reporting Standards (CHEERS) checklist was applied to evaluate the quality of the studies. Then cost and effectiveness data were extracted and summarized in a narrative way. Fifteen economic evaluations of RDT compared to other diagnostic methods were identified. The overall quality of studies varied greatly but most of them were scored to be of high or moderate quality. Ten of the fifteen studies reported that RDT was likely to be a cost-effective approach compared to its comparisons, but the results could be influenced by the alternatives, study perspectives, malaria prevalence, and the types of RDT. CONCLUSIONS: Based on available evidence, RDT had the potential to be more cost-effective than either microscopy or presumptive diagnosis. Further research is also required to draw a more robust conclusion.

The challenge of maintaining microscopist capacity at basic levels for malaria elimination in Jiangsu Province, China.

BACKGROUND: Local malaria transmission has decreased rapidly since the National Malaria Elimination Action Plan was launched in China in 2010. However, imported malaria cases from Africa and Southeast Asia still occur in China due to overseas laborers. Diagnosis by microscopy is the gold standard for malaria and is used in most hospitals in China. However, the current capacity of microscopists to manage malaria cases in hospitals and public health facilities to meet the surveillance needs to eliminate and prevent the reintroduction of malaria is unknown. METHODS: Malaria diagnoses were assessed by comparing the percentage of first visit and confirmed malaria diagnoses at Centers for Disease Control and Prevention (CDCs) and hospitals. The basic personnel information for public health departments and hospitals at different levels was investigated. The skills of microscopists for blood smear preparation and slide interpretation were also examined at the county and township levels. RESULTS: Inaccurate rate with 13.49% and 7.32%, respectively, in 2013 and 2014, from 341 and 355 reported cases from sub-provincial levels in Jiangsu province. Most of the 523 malaria cases reported in Nantong Prefecture from 2000 to 2014 involved patients who first visited county CDCs seeking treatment, however, none of these cases received confirmed diagnosis of malaria in townships or villages.The staff at county CDCs and hospitals with a higher education background performed better at making and interpreting blood smears than staff from townships. CONCLUSIONS: The network for malaria elimination in an entire province has been well established. However, an insufficient capacity for malaria diagnosis was observed, especially the preparing and reading the blood smears at the township and village levels, which is a challenge to achieving and maintaining malaria elimination.

Return of chloroquine sensitivity to Africa? Surveillance of African Plasmodium falciparum chloroquine resistance through malaria imported to China.

BACKGROUND: Chloroquine (CQ) was the cornerstone of anti-malarial treatment in Africa for almost 50 years, but has been widely withdrawn due to the emergence and spread of resistance. Recent reports have suggested that CQ-susceptibility may return following the cessation of CQ usage. Here, we monitor CQ sensitivity and determine the prevalence of genetic polymorphisms in the CQ resistance transporter gene (pfcrt) of Plasmodium falciparum isolates recently imported from Africa to China. METHODS: Blood samples were collected from falciparum malaria patients returning to China from various countries in Africa. Isolates were tested for their sensitivity to CQ using the SYBR Green I test ex vivo, and for a subset of samples, in vitro following culture adaptation. Mutations at positions 72-76 and codon 220 of the pfcrt gene were analyzed by sequencing and confirmed by PCR-RFLP. Correlations between drug sensitivity and pfcrt polymorphisms were investigated. RESULTS: Of 32 culture adapted isolates assayed, 17 (53.1%), 6 (18.8%) and 9 (28.1%) were classified as sensitive, moderately resistant, and highly resistant, respectively. In vitro CQ susceptibility was related to point mutations in the pfcrt gene, the results indicating a strong association between pfcrt genotype and drug sensitivity. A total of 292 isolates were typed at the pfcrt locus, and the prevalence of the wild type (CQ sensitive) haplotype CVMNK in isolates from East, South, North, West and Central Africa were 91.4%, 80.0%, 73.3%, 53.3% and 51.7%, respectively. The only mutant haplotype observed was CVIET, and this was almost always linked to an additional mutation at A220S. CONCLUSIONS: Our results suggest that a reduction in drug pressure following withdrawal of CQ as a first-line drug may lead to a resurgence in CQ sensitive parasites. The prevalence of wild-type pfcrt CQ sensitive parasites from East, South and North Africa was higher than from the West and Central areas, but this varied greatly between countries. Further surveillance is required to assess whether the prevalence of CQ resistant parasites will continue to decrease in the absence of widespread CQ usage.

The increasing importance of Plasmodium ovale and Plasmodium malariae in a malaria elimination setting: an observational study of imported cases in Jiangsu Province, China, 2011-2014.

BACKGROUND: Following initiation of China's National Malaria Elimination Action Plan in 2010, indigenous malaria infections in Jiangsu Province decreased significantly. Meanwhile imported Plasmodium infections have increased substantially, particularly Plasmodium ovale and Plasmodium malariae. Given the risk for malaria resurgence, there is an urgent need to understand the increase in imported P. ovale and P. malariae infections as China works to achieve national malaria elimination. METHODS: An observational study of imported malaria cases in Jiangsu Province, China was carried out for the period of 2011-2014. RESULTS: A total of 1268 malaria cases were reported in Jiangsu Province from 2011 to 2014. Although imported Plasmodium falciparum cases (n = 1058) accounted for 83.4 % of all reported cases in Jiangsu, P. ovale cases (14, 19, 30, and 46) and their proportion (3.7, 9.6, 8.8, and 13.0 %) of all malaria cases increased over the 4 years. Similarly, P. malariae cases (seven, two, nine, and 10) and proportion (1.9, 1.0, 2.6, and 2.8 %) of all malaria cases increased slightly during this time. A total of 98 cases of Plasmodium ovale curtisi (47/98, 48 %) and Plasmodium ovale wallikeri (51/98, 52 %) were identified as well. Latency periods were significant among these Plasmodium infections (p = 0.00). Also, this study found that the latency periods of P. ovale sp., P. malariae and Plasmodium vivax were significantly longer than P. falciparum. However, for both P. ovale curtisi and P. ovale wallikeri infections, the latency period analysis was not significant (p = 0.81). Misdiagnosis of both P. ovale and P. malariae was greater than 71.5 and 71.4 %, respectively. The P. ovale cases were misdiagnosed as P. falciparum (35 cases, 32.1 %), P. vivax (43 cases, 39.4 %) by lower levels of CDCs or hospitals. And, the P. malariae cases were misdiagnosed as P. falciparum (ten cases, 35.7 %), P. vivax (nine cases, 32.1 %) and P. ovale sp. (one case, 3.6 %). Geographic distribution of imported P. ovale sp. and P. malariae cases in Jiangsu Province mainly originated from sub-Saharan Africa such as Equatorial Guinea, Nigeria, and Angola. CONCLUSIONS: Although the vast majority of imported malaria cases were due to P. falciparum, the increase in other rare Plasmodium species originating from sub-Saharan Africa and Southeast Asia should be closely monitored at all levels of health providers focusing on diagnosis and treatment of malaria. In addition to a receptive vector environment, long latency periods and misdiagnosis of P. malariae and P. ovale sp. increase the risk of re-introduction of malaria in China.

Production of Plasmodium vivax enolase in Escherichia coli and its protective properties.

Plasmodium vivax predominates in South-East Asia and the American continent, causes significant morbidity and inflicts a huge socioeconomic burden. Sequencing completion of the Plasmodium vivax genome and transcriptome provides the chance to identify antigens. Enolase is the eighth enzyme in the glycolytic pathway, which, apart from its glycolytic function, also possess antigenic properties and is present on the cell wall of many invasive organisms, such as Candida albicans. In order to assess whether enolase of Plasmodium vivax is also antigenic, in this study, we first reported the expression and purification of recombinant Plasmodium vivax enolase (r-Pven) in Escherichia coli, using prokaryotic expression vector. The r-Pven was expressed in soluble form in E. coli, and the expression was verified by SDS-PAGE and western blotting analysis. The r-Pven was purified to 90% purity by nickel-nitrilotriacetic acid (Ni2+-NTA) resin chromatography. For reactivity with r-Pven, compared with the average values of the reactivity of control serum samples, the average values of the reactivity of 99 individual serums from vivax malaria patients appeared higher, and there was significant difference between them (p=0.0117<0.05). Mice anti-r-Pven antibodies inhibited the growth of in vitro cultures of P. falciparum. Mice immunized with r-Pven showed protection against a challenge with the mouse malarial parasite Plasmodium berghei. The antibodies raised against r-Pven were specific for Plasmodium and did not react to the host tissues. These observations established Plasmodium vivax enolase to be a potential protective antigen.