Detection and evolutionary characterization of arboviruses in mosquitoes and biting midges on Hainan Island, China, 2019-2023.

We conducted a large-scale survey of arboviruses in mosquitoes and biting midges to assess the presence and spread of mosquito-borne pathogens currently circulating on Hainan Island, China. RT-PCR assays were used to detect the arbovirus species, distribution, and infection rates in mosquitoes and biting midges. Cell inoculation and high throughput sequencing were performed to isolate the viruses and assemble full viral genomes. Phylogenetic analysis was conducted to identify the viral genotypes and evolutionary relationships with known viruses. During 2019-2023, 32,632 mosquitoes and 21,000 biting midges were collected from 14 of 18 cities/counties on Hainan Island. Japanese encephalitis virus (JEV) was detected in Culex mosquitoes from five cities/counties, where the minimum infection rate (MIR) was 1.6 (0.6-2.6) per 1,000 females tested. Tembusu virus (TMUV) was detected in Culex mosquitoes from three cities/counties with MIR1.0 (0-2.2) per 1,000. Getah virus (GETV) was detected in Armigeres mosquitoes from Qionghai city with MIR 7.1 (0-15.2) per 1,000. Oya virus (OYAV) and Bluetongue virus (BTV) were detected in biting midges from Wanning city with MIRs of 0.4 (0-1.2) and 0.1 (0-10.2) per 1,000, respectively. Three JEV strains were isolated and clustered within the genotype I group, which is presently the dominant genotype in China. Three TMUV strains were isolated for the first time on Hainan Island that belonged to Cluster 3. Three isolated GETVs were identified as Group 3. BTV was reported for the first time on Hainan Island, and the complete genome for one BTV strain was successfully assembled, which was classified as serotype 1 based on the sequences of segment 2. These results stress the need to develop adequate surveillance plan measures to better control the public health threat of arboviruses carried by mosquitoes and biting midges in local regions.

Development and evaluation of RT-qPCR assays for two neglected orthobunyaviruses: Oya virus and Ebinur Lake virus.

OBJECTIVES: Oya virus (OYAV) and Ebinur lake virus (EBIV) belong to the genus Orthobunyavirus within the Peribunyaviridae family, and both are recognized as the novel virus with potential threat to the animal or public health. Given their potential to cause outbreaks and their detection in diverse samples across different regions, the need for a reliable and efficient molecular detection method for OYAV and EBIV becomes imperative. METHODS: The S-segment of OYAV and EBIV was used for designing specific primer and probe sets, which were employed in a real-time reverse transcription quantitative PCR (RT-qPCR) assay. The analytical performance of these assays, encompassing specificity, sensitivity, reproducibility, and fitness for purpose, was thoroughly evaluated across various sample matrices. RESULTS: The developed RT-qPCR assays were very specific to their respective targets. Both assays were highly reproducible (%CV<3) and sensitive with the 95% limit of detection (LOD) of 0.80 PFU/mL for OYAV primer probe set and 0.37 PFU/mL for EBIV primer probe set. Furthermore, the assays fitness for purpose was good as it could detect the specific viruses in virus-spiked serum samples, virus-inoculated mosquito samples, field caught mosquitoes and biting midge samples. CONCLUSIONS: Our study has successfully developed specific, sensitive, and reliable RT-qPCR assays for the detection of OYAV and EBIV. These assays hold great promise for their potential application in clinical and field samples in the future.

Factors affecting the transmission of dengue fever in Haikou city in 2019.

In this study, due to multiple cases of dengue fever in two locations in Haikou, Hainan, several factors affecting the transmission of dengue fever in Haikou in 2019 were analyzed. It was found that dengue fever spread from two sites: a construction site, which was an epidemic site in Haikou, and the university, where only four confirmed cases were reported. Comparative analysis revealed that the important factors affecting the spread of dengue fever in Haikou were environmental hygiene status, knowledge popularization of dengue fever, educational background, medical insurance coverage and free treatment policy knowledge and active response by the government.

Factors Contributing to the Pre-Elimination of Malaria from Hainan Island, China, 1986-2009.

Malaria was endemic in Hainan Island, China, for a lengthy period before its elimination. Here, we aim to gain a better understanding of malaria elimination by assessing and quantifying the relative effects of longitudinal changes in specific antimalarial interventions, socioeconomic development approaches, and malaria vectors on malaria prevalence in Hainan during the 1986-2009 pre-elimination period. Annual data were collected on the incidence of malaria, the number of residents protected by drugs (RPD), the number of residents protected by vector control, the presence of Anopheles minimus and Anopheles dirus, the annual per capita income of rural residents, major cash crop (rubber plantation) areas, the literacy rate of adult rural residents, and the rate of reinforced concrete house construction in rural areas. Backward stepwise multiple linear regression models were developed to identify the factors associated with the annual malaria incidence (AMI). The AMI declined from 20.3 to 0.8 per 10,000 population from 1986 to 2009; this decrease was significantly associated with an increase in the number of RPD and improved literacy rate among rural adults. The results of this study implied that the sustained distribution of antimalarial drugs and increase in education levels in risk areas significantly impacted the reduction and elimination of malaria infection in Hainan. We suggest that these findings could be applicable to malaria-endemic areas in Southeast Asia with similar natural and socioeconomic environments to Hainan and where malaria incidence has decreased to a low level.

Surveillance and Control of Malaria Vectors in Hainan Province, China from 1950 to 2021: A Retrospective Review.

Malaria is a serious mosquito-borne tropical disease impacting populations in tropical regions across the world. Malaria was previously hyperendemic in Hainan Province. Due to large-scale anti-malarial intervention, malaria elimination in the province was achieved in 2019. This paper reviews the literature on the ecology, bionomics, and control of malaria vectors in Hainan from 1951 to 2021. We searched PubMed, and the China national knowledge infrastructure (CNKI) database for relevant articles published and included three other important books published in Chinese or English in order to summarize research on species, distribution, vectorial capacity, ecology, the resistance of malaria vectors to insecticides, and malaria vector control in Hainan Province. A total of 239 references were identified, 79 of which met the criteria for inclusion in our review. A total of six references dealt with the salivary gland infection of Anophelines, six with vectorial capacity, 41 with mosquito species and distribution, seven with seasonality, three with blood preference, four with nocturnal activity, two with flight distance, 13 with resistance to insecticides, and 14 with vector control. Only 16 published papers met the criteria of addressing malaria vectors in Hainan over the last 10 years (2012-2021). Anopheles dirus and Anopheles minimus are primary malaria vectors, mainly distributed in the southern and central areas of Hainan. Indoor residual spraying with DDT and the use of ITNs with pyrethroid insecticides were the main interventions taken for malaria control. Previous studies on ecology, bionomics, and resistance of vectors provided scientific evidence for optimizing malaria vector control and contributed to malaria elimination in Hainan Province. We hope our study will contribute to preventing malaria reestablishment caused by imported malaria in Hainan. Research on malaria vectors should be updated to provide scientific evidence for malaria vector control strategies post-elimination as the ecology, bionomics, and resistance of vectors to insecticides may change with changes in the environment.

Prevalence of glucose-6-phosphate dehydrogenase deficiency (G6PDd) and clinical implication for safe use of primaquine in malaria-endemic areas of Hainan Province, China.

Primaquine, the only licensed antimalarial drug for eradication of Plasmodium vivax and Plasmodium ovale malaria, may cause acute hemolytic anemia in individuals with glucose-6-phosphate dehydrogenase deficiency (G6PDd) during treatment. The different prevalence and distribution patterns of G6PDd in Hainan, the ancient malaria-endemic area, are unclear. This study included 5,622 suspected malaria patients between 2009 and 2011 in 11 counties of Hainan. Glucose-6-phosphate dehydrogenase deficiency prevalence was determined using the fluorescent spot test (FST) and malaria patients was confirmed by a positive light microscopy. The G6PDd prevalence for different ethnic groups, genders, and counties were calculated and compared using χ2-test. Spatial cluster and Spearman rank correlation of G6PDd prevalence and malaria incidence were analyzed. The overall G6PDd prevalence of study population was 7.45%. The G6PDd prevalence of males, Li ethnic minority, and malaria patients was significantly higher than that of females, Han ethnic majority, and non-malarial patients (p < 0.01), respectively. The spatial cluster of G6PDd and malaria located in south-western and central-southern Hainan, respectively, with no significant correlation. The study provides essential information on G6PDd prevalence in ancient malaria-endemic areas of Hainan Province. We also highlight the need for a better understanding of the mechanisms underlying the relationship between G6PDd prevalence and malaria incidence. These findings provide a reference for the safety of the primaquine-based intervention, even after malaria elimination.

Malaria elimination on Hainan Island despite climate change.

Background: Rigorous assessment of the effect of malaria control strategies on local malaria dynamics is a complex but vital step in informing future strategies to eliminate malaria. However, the interactions between climate forcing, mass drug administration, mosquito control and their effects on the incidence of malaria remain unclear. Methods: Here, we analyze the effects of interventions on the transmission dynamics of malaria (Plasmodium vivax and Plasmodium falciparum) on Hainan Island, China, controlling for environmental factors. Mathematical models were fitted to epidemiological data, including confirmed cases and population-wide blood examinations, collected between 1995 and 2010, a period when malaria control interventions were rolled out with positive outcomes. Results: Prior to the massive scale-up of interventions, malaria incidence shows both interannual variability and seasonality, as well as a strong correlation with climatic patterns linked to the El Nino Southern Oscillation. Based on our mechanistic model, we find that the reduction in malaria is likely due to the large scale rollout of insecticide-treated bed nets, which reduce the infections of P. vivax and P. falciparum malaria by 93.4% and 35.5%, respectively. Mass drug administration has a greater contribution in the control of P. falciparum (54.9%) than P. vivax (5.3%). In a comparison of interventions, indoor residual spraying makes a relatively minor contribution to malaria control (1.3%-9.6%). Conclusions: Although malaria transmission on Hainan Island has been exacerbated by El Nino Southern Oscillation, control methods have eliminated both P. falciparum and P. vivax malaria from this part of China.

Erratum: Author Correction: Malaria elimination on Hainan Island despite climate change.

[This corrects the article DOI: 10.1038/s43856-022-00073-z.].

The relationship between rising temperatures and malaria incidence in Hainan, China, from 1984 to 2010: a longitudinal cohort study.

BACKGROUND: The influence of rising global temperatures on malaria dynamics and distribution remains controversial, especially in central highland regions. We aimed to address this subject by studying the spatiotemporal heterogeneity of malaria and the effect of climate change on malaria transmission over 27 years in Hainan, an island province in China. METHODS: For this longitudinal cohort study, we used a decades-long dataset of malaria incidence reports from Hainan, China, to investigate the pattern of malaria transmission in Hainan relative to temperature and the incidence at increasing altitudes. Climatic data were obtained from the local meteorological stations in Hainan during 1984-2010 and the WorldClim dataset. A temperature-dependent R0 model and negative binomial generalised linear model were used to decipher the relationship between climate factors and malaria incidence in the tropical region. FINDINGS: Over the past few decades, the annual peak incidence has appeared earlier in the central highland regions but later in low-altitude regions in Hainan, China. Results from the temperature-dependent model showed that these long-term changes of incidence peak timing are linked to rising temperatures (of about 1·5°C). Further, a 1°C increase corresponds to a change in cases of malaria from -5·6% (95% CI -4·5 to -6·6) to -9·2% (95% CI -7·6 to -10·9) from the northern plain regions to the central highland regions during the rainy season. In the dry season, the change in cases would be 4·6% (95% CI 3·7 to 5·5) to 11·9% (95% CI 9·8 to 14·2) from low-altitude areas to high-altitude areas. INTERPRETATION: Our study empirically supports the idea that increasing temperatures can generate opposing effects on malaria dynamics for lowland and highland regions. This should be further investigated and incorporated into future modelling, disease burden calculations, and malaria control, with attention for central highland regions under climate change. FUNDING: Scientific and Technological Innovation 2030: Major Project of New Generation Artificial Intelligence, National Natural Science Foundation of China, Beijing Natural Science Foundation, National Key Research and Development Program of China, Young Elite Scientist Sponsorship Program by CAST, Research on Key Technologies of Plague Prevention and Control in Inner Mongolia Autonomous Region, and Beijing Advanced Innovation Program for Land Surface Science.

MicroRNA-186 is associated with hypoxia-inducible factor-1α expression in chronic obstructive pulmonary disease.

BACKGROUND: MicroRNAs (miRNAs) are a family of small noncoding RNAs and are essential in the regulation of gene expression. Their impacts on gene expression have been reported in various diseases. The role of hypoxia-inducible factor 1 alpha (HIF-1α) in the development and progression of chronic obstructive pulmonary disease (COPD) has also been demonstrated. However, the role of microRNA-186 (miR-186) in relation to HIF in COPD is unknown. METHODS: Cell culture experiments were performed using human lung fibroblast cells (MRC-5). Cell viability was determined by MTT and flow cytometry assays. Reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blot analysis were used to assess the expression levels of HIF-1α and inflammatory cytokines. Dual-luciferase reporter assays were used to reveal the correlation between miR-186 and HIF-1α. RESULTS: After miR-186 transfection, the cell lines showed reduced proliferation and increased apoptosis. After overexpression of miR-186, we found that the HIF-1α expression level was reduced in MRC-5 cells. We found that miR-186 can affect apoptosis of inflammatory fibroblasts through the regulation of HIF-1α and affect the downstream signaling pathways. CONCLUSIONS: These data suggested that miR-186 contributes to the pathogenesis of COPD and that miRNA-186 may also affect the HIF-1α-dependent lung structure maintenance program.

Telomere length, ZNF208 genetic variants and risk of chronic obstructive pulmonary disease in the Hainan Li population.

BACKGROUND: Chronic obstructive pulmonary disease (COPD) is a disease characterized by airflow limitation. It is not completely reversible and shows progressive development. ZNF208 rs8105767 affects telomere length, although the impact of telomere on COPD is still controversial. In the present study, we aimed to explore the impact of the ZNF208 gene polymorphism on telomere length and also that of telomere length on COPD in the Hainan Li population. METHODS: In total, 270 COPD patients and 288 controls were recruited. Telomere length was measured by a quantitative real-time polymerase chain reaction. Five single nucleotide polymorphisms in ZNF208 were selected and genotyping was performed using MassARRAY software (Agena Bioscience Co. Ltd, San Diego, CA, USA). Differences in telomere length among the subjects with three genotypes of related genes were assessed using analysis of variance. Unconditional logistic regression was used to calculate odds ratios (OR) as the indicator of association between telomere length and COPD risk. RESULTS: Relative telomere length in the COPD group and control group was 0.66 ± 0.47 and 1.44 ± 0.89, respectively. We grouped according to a median of 0.8284 for telomere length and observed that the risk of COPD for individuals with a telomere length less than 0.8284 is 2.92 times that for individuals with a telomere length longer than 0.8284 (OR = 2.92, 95% confidence interval = 2.01-4.25, p = 1.91 × 10-8 ). Subjects carrying the G allele of rs2188972 had a longer telomere length. Subjects carrying the carrying the CA genotype of rs8103163 and AC genotype of rs7248488 had a longer telomere length compared to wild-type individuals. CONCLUSIONS: Shorter telomeres increase COPD risk and the ZNF208 polymorphism affects telomere length in COPD patients.

Prevalence and risk factors of chronic obstructive pulmonary diseases in a Hlai community in Hainan Island of China.

OBJECTIVES: We investigated prevalence and risk factors of chronic obstructive pulmonary disease (COPD) in a population of Hlai (the Li) ethnicity, a major minority, in Qicha Town, Changjiang County, Hainan Province, PRC, during 2014. METHODS: All residents at the age of 40 years or older were interviewed with standardized questionnaires. Spirometry was performed to measure the possible airflow limitation. According to the GOLD criteria, post bronchodilator FEV1/FVC < 70% was defined as COPD. Case-control study was used to screen the risk factors by analyzing COPD group (212 cases) and non-COPD control group (236 cases). Single factor analysis and multiple factor logistic regression analysis were used as statistical methods. RESULTS: The prevalence of COPD in the residents at the age of 40 years or older of Hlai community was 5.07% (286/5637) (95% CI = 0.045-0.057). In the logistic regression analysis, the COPD prevalence was 5.07% (147/2901) in men and 5.08% (139/2736) in women, respectively, with odds ratio (OR) 1.003, 95% CI 0.790-1.272 and P > 0.05, suggesting that the sex did not affect the COPD prevalence in the investigated samples, but age (OR = 1.096), expectoration (OR = 87.917), locomotor activity limitation (OR = 3.908) and frequency of respiration (OR = 2.512) were risk factors and associated with the development of COPD. Notably, although the tobacco smoker in male and female COPD patients were 48.6% (54/111) and 4.0% (4/101), respectively, passive smokers in female with COPD were 45.6% (46/101). CONCLUSION: In the Hlai population aged ≥40 years, the COPD prevalence was 5.07%. Smoking, age, expectoration, locomotor activity limitation and frequency of respiration were risk factors of COPD in Hlai ethnicity.

Extensive Resistance of Anopheles sinensis to Insecticides in Malaria-Endemic Areas of Hainan Province, China.

Anopheles sinensis is one of the major malaria vectors and among the dominant species in Hainan Province, China. The resistance of An. sinensis to insecticides is an important threat to malaria control. However, few reports on insecticide resistance of An. sinensis were reported in this area. Eight districts in Hainan Province were selected as the study areas. Insecticide susceptibility bioassays were tested on wild-caught female mosquitoes of An. sinensis to 4% dichlorodiphenyltrichloroethane (DDT), 0.05% deltamethrin, and 5% malathion by using the World Health Organization standard resistance tube assay procedure. All the tested An. sinensis mosquitoes demonstrated resistance to 4% DDT, with less than 72% mortality in the standard assay. The populations from Baisha and Qiongzhong demonstrated possible resistance to 0.05% deltamethrin, with 94-95% mortality, whereas the populations from other districts demonstrated resistance to 0.05% deltamethrin in the standard assay. The populations from Baisha, Qiongzhong, and Dongfang demonstrated susceptibility to 5% malathion, but the populations from other districts demonstrated resistance. These results facilitate the improvement of effective control strategies for malaria vector mosquitoes in Hainan.

A Cost-Effectiveness Analysis of Plasmodium falciparum Malaria Elimination in Hainan Province, 2002-2012.

In Hainan Province, China, great achievements in elimination of falciparum malaria have been made since 2010. There have been no locally acquired falciparum malaria cases since that time. The cost-effectiveness of elimination of falciparum malaria has been analyzed in Hainan Province. There were 4,422 falciparum malaria cases reported from 2002 to 2012, more cases occurred in males than in females. From 2002 to 2012, a total of 98.5 disability-adjusted life years (DALYs) were reported because of falciparum malaria. Populations in the age ranges of 15-25 and 30-44 years had higher incidences and DALYs than other age groups. From 2002 to 2012, malaria-related costs for salaries of staff, funds from the provincial government, national government, and the GFATM were US$3.02, US$2.24, US$1.44, and US$5.08 million, respectively. An estimated 9,504 falciparum malaria cases were averted during the period 2003-2012. The estimated cost per falciparum malaria case averted was US$116.5. The falciparum malaria elimination program in Hainan was highly effective and successful. However, funding for maintenance is still needed because of imported cases.

Sensitivity of Plasmodium falciparum to antimalarial drugs in Hainan Island, China.

Pyronaridine and artesunate have been shown to be effective in falciparum malaria treatment. However, pyronaridine is rarely used in Hainan Island clinically, and artesunate is not widely used as a therapeutic agent. Instead, conventional antimalarial drugs, chloroquine and piperaquine, are used, explaining the emergence of chloroquine-resistant Plasmodium falciparum. In this article, we investigated the sensitivity of P. falciparum to antimalarial drugs used in Hainan Island for rational drug therapy. We performed in vivo (28 days) and in vitro tests to determine the sensitivity of P. falciparum to antimalarial drugs. Total 46 patients with falciparum malaria were treated with dihydroartemisinin/piperaquine phosphate (DUO-COTECXIN) and followed up for 28 day. The cure rate was 97.8%. The mean fever clearance time (22.5 ± 10.6 hr) and the mean parasite clearance time (27.3 ± 12.2 hr) showed no statistical significance with different genders, ages, temperatures, or parasite density (P > 0.05). The resistance rates of chloroquine, piperaquine, pyronarididine, and artesunate detected in vitro were 71.9%, 40.6%, 12.5%, and 0%, respectively (P < 0.0001). The resistance intensities decreased as follows: chloroquine > piperaquine > pyronarididine > artesunate. The inhibitory dose 50 (IC50) was 3.77 × 10(-6) mol/L, 2.09 × 10(-6) mol/L, 0.09 × 10(-6) mol/L, and 0.05 × 10(-6) mol/L, and the mean concentrations for complete inhibition (CIMC) of schizont formation were 5.60 × 10(-6) mol/L, 9.26 × 10(-6) mol/L, 0.55 × 10(-6) mol/L, and 0.07 × 10(-6) mol/L, respectively. Dihydroartemisinin showed a strong therapeutic effect against falciparum malaria with a low toxicity.

Cytochrome P450 2C9 (CYP2C9) polymorphisms in Chinese Li population.

BACKGROUND: The frequencies of Cytochrome P450 2C9 (CYP2C9) genotypes were various between populations. The aim of this study was to investigate the frequencies of the major variants of the CYP2C9 in Chinese Li minority populations. METHODS: The promoter, exons and surrounding introns, and 3'-untranslated region of the CYP2C9 gene was detected by DNA sequencing to investigate in 100 unrelated healthy Chinese Li subjects. The protein function prediction was used the online tools: Sorting Intolerant From Tolerant (SIFT) and Phenotyping Version 2 (PolyPhen-2). The comparison of CYP2C9 allele frequencies in different populations were analyzed by Chi-square (χ(2)) test. Linkage disequilibrium (LD) analysis was performed using Haploview software. RESULTS: We identified 17 different CYP2C9 single nucleotide polymorphisms (SNPs) in the Li population, including two missense mutations (3549 G > A and 42614 A > C) and two silent mutations (3514 T > Cand 50298A > T). The protein function prediction revealed the two missense mutations result in protein damaging. In addition, we detected the allele frequencies of CYP2C9*1, CYP2C9*3 and CYP2C9*42 were 98%, 1%, and 1%, respectively. Finally, we compared three major allelic frequency (CYP2C9*1, CYP2C9*2, and CYP2C9*3) between Li and other populations. We found that our results were similar to East Asians and Africans.

Eliminating Plasmodium falciparum in Hainan, China: a study on the use of behavioural change communication intervention to promote malaria prevention in mountain worker populations.

BACKGROUND: In the island of Hainan, the great majority of malaria cases occur in mountain worker populations. Using the behavioral change communication (BCC) strategy, an interventional study was conducted to promote mountain worker malaria prevention at a test site. This study found the methods and measures that are suitable for malaria prevention among mountain worker populations. METHODS: During the Plasmodium falciparum elimination stage in Hainan, a representative sampling method was used to establish testing and control sites in areas of Hainan that were both affected by malaria and had a relatively high density of mountain workers. Two different methods were used: a BCC strategy and a conventional strategy as a control. Before and after the intervention, house visits, core group discussions, and structural surveys were utilized to collect qualitative and quantitative data regarding mountain worker populations (including knowledge, attitudes, and practices [KAPs]; infection status; and serological data), and these data from the testing and control areas were compared to evaluate the effectiveness of BCC strategies in the prevention of malaria. RESULTS: In the BCC malaria prevention strategy testing areas, the accuracy rates of malaria-related KAP were significantly improved among mountain worker populations. The accuracy rates in the 3 aspects of malaria-related KAP increased from 37.73%, 37.00%, and 43.04% to 89.01%, 91.53%, and 92.25%, respectively. The changes in all 3 aspects of KAP were statistically significant (p < 0.01). In the control sites, the changes in the indices were not as marked as in the testing areas, and the change was not statistically significant (p > 0.05). Furthermore, in the testing areas, both the percentage testing positive in the serum malaria indirect fluorescent antibody test (IFAT) and the number of people inflicted decreased more significantly than in the control sites (p < 0.01). CONCLUSION: The use of the BCC strategy significantly improved the ability of mountain workers in Hainan to avoid malarial infection. Educational and promotional materials and measures were developed and selected in the process, and hands-on experience was gained that will help achieve the goal of total malaria elimination in Hainan.

[Resistance of Anopheles sinensis to three common insecticides in Hainan Province].

Anopheles sinensis adults were collected by cow-baited trap in Haikou City, Sanya City, Lingshui County, Changjiang County and Qiongzhong County of Hainan Province during 2011-2013. The mosquitoes were exposed to insecticide impregnated papers with discriminating concentrations of DDT (4%), deltamethrin (0.05%), and malathion (5%) using WHO standard assays. Knockdown rate was recorded at 10, 15, 20, 30, 40, 50, and 60 min, and KT50 values were calculated. Mortality was recorded after 24 hours of exposure. The resistance level was graded as sensitive group (S) with a mortality rate of 98%-100%, preliminary resistance group (M) with mortality rate of 80%-97%, and resistance group (R) with mortality rate of lower than 80%. The results showed that the mortality rate of An. sinensis in Qiongzhong County in 24 h-post-exposure to 0.05% deltamethrin was 95.0% with a resistance degree of M. That to 0.05% deltamethrin in the other 4 sites was 17.0%-63.0%, all with a resistance degree of R. That to 4% DDT in Haikou, Sanya, Lingshui, Qiongzhong and Changjiang was 36.0%, 27.0%, 24.0%, 59.1%, and 82.0%, with a resistance level of R, R, R, R, and M, respectively. That to 5% malathion in Haikou, Sanya, and Lingshui was 16.0%-41.0%, all with a resistance degree of R, while that to malathion in Qiongzhong and Changjiang was 100% and 98.0%, respectively, with a resistance level of S.

A case of Plasmodium ovale wallikeri infection in a Chinese worker returning from West Africa.

In contrast to the gradual reduction in the number of locally transmitted malaria cases in China, the number of imported malaria cases has been increasing since 2008. Here, we report a case of a 39-year-old Chinese man who acquired Plasmodium ovale wallikeri infection while staying in Ghana, West Africa for 6 months in 2012. Microscopic examinations of Giemsa-stained thin and thick blood smears indicated Plasmodium vivax infection. However, the results of rapid diagnostic tests, which were conducted 3 times, were not in agreement with P. vivax. To further check the diagnosis, standard PCR analysis of the small-subunit rRNA gene was conducted, based on which a phylogeny tree was constructed. The results of gene sequencing indicated that this malaria is a variant of P. ovale (P. ovale wallikeri). The infection in this patient was not a new infection, but a relapse of the infection from the one that he had contracted in West Africa.

[Resistance assay of malaria vectors to four kinds of common insecticides in some endemic areas of Hainan Province].

OBJECTIVE: To determine the resistance of malaria vectors to four kinds of common insecticides in some endemic areas of Hainan Province. METHODS: Anopheline mosquitoes were collected between 2008 and 2010 from malaria endemic areas where insecticides were used for years. Anopheles dirus were collected from human-baited trap in Wangxia Town of Changjiang County. An. minimus and An. sinensis were collected by cow-baited trap in Jiangbian Town of Dongfang City. F0 generation female An. sinensis, F1 generation of female An. dirus and An. minimus were selected and exposed to insecticide impregnated papers with discriminating concentrations of DDT (4%), deltamethrin (0.05%), cyfluthrin (0.15%), and malathion (5%) using WHO standard assays. Knockdown rate was recorded at 10, 15, 20, 30, 40, 50, and 60 min, and KTm values were calculated. Mortality was recorded after 24 hours of exposure. RESULTS: Mortality in An. dirus was 100% to DDT, deltamethrin and malathion. Knockdown rate of An. dirus exposed to DDT and deltamethrin was 82.0% and 100%, with a KT50 value of 46.9 and 18.4 min, respectively. Mortality of An. minimus to DDT, deltamethrin, cyfluthrin and malathion was 98.1%, 99.0%, 100%, and 100%, respectively. The knockdown rate of An. minimus to DDT, deltamethrin, and cyfluthrin was 96.3%, 99.0%, and 100%, respectively, and the KTs value was 31.3, 16.8, and 7.4 min, respectively. Mortality of An. sinensis to DDT, deltamethrin, and malathion was 19.8%, 22.9%, and 43.8%, respectively. Knockdown rate of An. sinensis to DDT and deltamethrin was 2.0%, the KT50 can not be calculated. CONCLUSION: An. dirus and An. minimus, the main malaria vectors in the survey sites of Hainan Province, are susceptible to the four insecticides, while secondary malaria vector An. sinensis showed resistance to DDT, deltamethrin, and malathion.