登革热的传播模式、蚊媒及防控

在自然界存在两种登革热传播模式:人-伊蚊-人循环,蚊媒是埃及伊蚊与白纹伊蚊。猴-伊蚊-猴循环,蚊媒是白纹伊蚊与白雪伊蚊群。我国学者首先于1975年从无输入性病例的我国西南边疆山林地区的白纹伊蚊体内分离到登革热病毒4型,白纹伊蚊承担两种传播模式的中介。本研究介绍了埃及伊蚊与白纹伊蚊的生态习性与全球及在中国的分布。认为在我国厦门地区迄今为止还未曾发现过埃及伊蚊的存在,也简介了沃尔巴克体新技术防控蚊媒研究的进展。     

本期重点推介

正登革病毒(dengue virus)是一种流行广泛的蚊媒病毒,主要经伊蚊Aedes传播,可导致登革热等重大疾病。由于长期使用杀虫剂对蚊媒进行防控,使蚊虫产生了抗药性。为了检验蚊虫抗药性对其感染和传播病毒的影响,南方医科大学公共卫生学院贾志荣和陈晓光等筛选建立了白纹伊蚊Aedes albopictus抗溴氰菊酯的实验室品系,并利用RT-PCR和RTq PCR检测了感染登革病毒后0,4,7和10 d白纹伊蚊溴氰     

白纹伊蚊幼虫期经香茅液处理后成蚊刺叮选择行为的变化

植物性驱蚊剂的开发研究受到了极大的关注,但昆虫的习惯化行为反应是制约有效植物性驱蚊剂发展的障碍之一.为了了解登革热重要媒介昆虫白纹伊蚊Aedes albopictus对植物性驱避剂的行为机制,本研究在分析了香茅油溶液对幼虫存活影响和对成蚊驱避作用的基础上,将白纹伊蚊幼虫于经稀释的香茅Cymbopogon nardus油溶液中饲养,然后通过局部涂肤的方法,测定白纹伊蚊雌蚊的刺叮选择行为变化.结果显示,香茅油对成蚊具有显著的驱避效果,但是幼虫经香茅油溶液饲养后,尽管仍具有显著的驱避作用,但是44.5 mg/L和178 mg/L的低浓度香茅油对白纹伊蚊的驱避作用发生了明显的减弱.说明幼虫期对精油溶液的经历,可诱导白纹伊蚊刺叮选择行为的习惯化反应,从而降低植物性驱避剂的驱避效果.     

登革2型病毒E蛋白结构域Ⅲ的表达及多克隆抗体制备

登革病毒(Dengue virus, DENV)属于黄病毒科(Flaviviridae),黄病毒属(Flavivirus),为单股正链RNA病毒,有4个不同的血清型(DENV- 1,2,3,4),主要通过埃及伊蚊(Aedes aegypti)和白纹伊蚊(Aedes albopictus)传播,可引起登革热、登革出血热、登革休克综合征等多种疾病 [1,2].     

我国白纹伊蚊对三种拟除虫菊酯类杀虫剂抗性检测诊断剂量的建立

【目的】建立中国白纹伊蚊Aedes albopictus成蚊对溴氰菊酯、氯菊酯和高效氯氟氰菊酯杀虫剂抗性检测的诊断剂量。【方法】应用溴氰菊酯、氯菊酯和高效氯氟氰菊酯原药制作不同浓度的药膜滤纸,接触筒法测定白纹伊蚊实验室敏感品系成蚊对3种拟除虫菊酯类杀虫剂的敏感性,记录1 h的击倒数和24 h的死亡数。应用Excel 2007和SPSS20.0进行数据统计处理,并制作杀虫剂的毒力回归线,计算各自的LC_(50)和LC_(99)值。以2倍LC_(99)值作为区分抗性和敏感种群的诊断剂量,制作药膜滤纸,接触筒法生物测定海口市白纹伊蚊现场种群成蚊对杀虫剂的抗药性。【结果】溴氰菊酯、高效氯氟氰菊酯和氯菊酯杀虫剂对白纹伊蚊实验室敏感品系成蚊的LC_(50)值分别为0.00619%,0.01403%和0.05009%,LC_(99)值分别为0.05175%,0.11859%,和0.53165%,相对应的诊断剂量分别为0.1035%,0.2372%和1.0633%。应用上述溴氰菊酯、氯菊酯和高效氯氟氰菊酯诊断剂量测定的海口市白纹伊蚊现场种群的死亡率分别为22.58%,36.29%和40.83%,表明该种群对这3种菊酯类杀虫剂均已产生了抗性。【结论】本研究建立的白纹伊蚊对3种拟除虫菊酯类杀虫剂的诊断剂量可作为该蚊成蚊抗药性监测的参考。     

白纹伊蚊和埃及伊蚊对基孔肯雅病毒的易感性和传播性的研究

用４株基孔肯雅病毒经口感染白纹伊蚊和埃及伊蚊,进行了易感性和传播性的研究。结果表明,这两种蚊虫对基孔肯雅病毒易感。无论白纹伊蚊或埃及伊蚊,感染后第５－６天即可通过吸血将病毒传播给乳鼠,至第８－１３天,传播率可高达５５．５５％－１００％。感染蚊亦可经叮咬将病毒传播给小鸡。埃及伊蚊的易感性和传播率高于白纹伊蚊。实验还发现,不同来源毒株之间存在一定差异,如分离自云南白纹伊蚊的Ｍ８１株的感染率和传播率均高于其它毒株。这些结果表明,白纹伊蚊和埃及伊蚊在基孔肯雅病毒的保存和传播中起重要作用。     

豫南罗索线虫防制轮胎积水内白纹伊蚊的试验

白纹伊蚊Aedes albopictus是我国和东南亚地区最常见的蚊种之一,分布面广,与人关系密切,为乙型脑炎和登革热的传播媒介。防制白纹伊蚊对消除此两种病的流行和传播具有重要意义。汽车的废旧轮胎多堆放于露天场所,雨后积水成为白纹伊蚊适宜的孳生地,附近居民深受其害,为此选择了某厂区轮胎积水环境投放豫南罗索线虫(Rommanomermis yunanensis)作     

登革2型病毒E蛋白结构域Ⅲ的表达及多克隆抗体制备

登革病毒（Dengue virus,DENV）属于黄病毒科（Flaviviridae）,黄病毒属（Flavivirus）,为单股正链RNA病毒,有4个不同的血清型（DENV-1,2,3,4）,主要通过埃及伊蚊（Aedes aegypti）和白纹伊蚊（Aedes albopictus）传播,可引起登革热、登革出血热、登革休克综合征等多种疾病。     

中国白纹伊蚊亚组伊蚊的研究——Ⅰ.成蚊

白纹伊蚊亚组(Aedes albopictus subgroup)是复蚊亚属(Stegomyia)伊蚊的重要类群之一,例如其中的白纹伊蚊是我国登革热的媒介之一,而且在福建、四川等地,它被认为与乙型脑炎的传播有关。本亚组所属种类,尤其是它们的幼虫和蛹,都很近似,因而过去有些种类不易区分。最近黄耀民(Huang,1972a;1979)对东南亚的本亚组蚊种作了比较详细的分类研究,才澄清了一些问题,提高了这方面的水平。     

AalDV-3介导的miRNA海绵载体构建及其在埃及伊蚊体内外的抑制效果测试

miRNA不仅广泛参与了蚊生长、发育的调控,还参与了媒介与病原体的相互作用,因此miRNAs可能作为蚊虫防制与蚊媒病原体控制的靶点,但目前尚无有效的转导系统。为研究白纹伊蚊浓核病毒-3(AalDV-3)是否可以作为有效的载体,我们利用内含子内表达miRNA海绵的策略,以埃及伊蚊内源性miR-210为靶基因,构建了重组病毒AalDV-anti-210。使用跨内含子引物通过RT-PCR验证了内含子可在蚊体内外的剪接效果,通过qPCR方法对埃及伊蚊Aag2细胞与埃及伊蚊幼虫的感染后的miR-210的表达水平进行检测。结果证实,含有miRNA海绵的内含子可以有效的蚊体内外剪接,重组病毒可以在蚊体内外高效的抑制靶miR-210的表达水平。该结果为探索AalDV-3在蚊虫基因功能研究及生物杀虫剂方面的潜在价值提供了理论依据。     

Resistance of Aedes aegypti from the state of São Paulo,Brazil, to organophosphates insecticides

Since the reintroduction of Aedes aegypti in the state of S o Paulo, in the middle of the 1980-decade, organophosphate insecticides are being used to control the dengue vector. In 1996, an annual program for monitoring the susceptibility of Ae. aegypti to the insecticides was implemented. Some of the results of this monitoring program are presented. Ae. aegypti populations from ten localities have been submitted to bioassays with the diagnostic dose of temephos and fenitrothion. Only two (Mar lia and Presidente Prudente) remain susceptible to both insecticides and one (Santos) exhibits true resistance. Ae. aegypti from the remaining localities showed an incipient altered susceptibility. Resistance ratios varied from 1.2 to 2.9 for temephos and from 1.5 to 3.2 to fenitrothion, indicating moderate levels of resistance. Biochemical assays did not detect alterations in the enzyme acetilcholinesterase, but indicated that resistance is associated with esterases.     

Larval competition alters susceptibility of adult Aedes mosquitoes to dengue infection

Dengue, the most important human arboviral disease, is transmitted primarily by Aedes aegypti and, to a lesser extent, by Aedes albopictus. The current distributions of these invasive species overlap and are affected by interspecific larval competition in their container habitats. Here we report that competition also enhances dengue infection and dissemination rates in one of these two vector species. We determined the effects of competition on adult A. aegypti and A. albopictus, comparing their susceptibility to infection with a Southeast Asian strain of dengue-2 virus. High levels of intra- or interspecific competition among larvae enhanced the susceptibility of A. albopictus to dengue virus infection and potential for transmission, as indicated by disseminated infections. Doubling the number of competing larvae (A. albopictus or A. aegypti), led to a significant (more than 60%) increase in the proportion of A. albopictus with disseminated dengue-2 infection. Competition-enhanced vector competence appears to result from a reduction in 'barriers' (morphological or physiological) to virus infection and dissemination and may contribute to the importance of A. albopictus in dengue transmission. Similar results for other unrelated arboviruses suggest that larval competition, common in mosquitoes, should be considered in estimates of vector competence for pathogens that infect humans.     

The efficacy of a chitin synthesis inhibitor against field populations of organophosphate-resistant Aedes aegypti in Brazil

The mosquito Aedes aegypti is the main focus of dengue control campaigns. Because of widespread resistance against conventional chemical insecticides, chitin synthesis inhibitors (CSIs) are considered control alternatives. We evaluated the resistance status of four Brazilian Ae. aegypti populations to both the organophosphate temephos and the pyrethroid deltamethrin, which are used in Brazil to control larvae and adults, respectively. All vector populations exhibited high levels of temephos resistance and varying rates of alterations in their susceptibility to pyrethroids. The effect of the CSI novaluron on these populations was also investigated. Novaluron was effective against all populations under laboratory conditions. Field-simulated assays with partial water replacement were conducted to evaluate novaluron persistence. Bioassays were continued until an adult emergence inhibition of at least 70% was attained. We found a residual effect of eight weeks under indoor conditions and novaluron persisted for five-six weeks in assays conducted in an external area. Our data show that novaluron is effective against the Ae. aegypti populations tested, regardless of their resistance to conventional chemical insecticides.     

Association of insecticide use and alteration on Aedes aegypti susceptibility status

Dengue and dengue hemorrhagic fever, vector-borne diseases transmitted by the mosquito Aedes aegypti, are presently important public health problems in Brazil. As the strategy for disease control is based on vector control through the use of insecticides, the development of resistance is a threat to programs efficacy. The objective of this study was to compare the Aedes aegypti susceptibility in nine vector populations from the state of S?o Paulo and seven from Northeast region of Brazil, since there was a difference on group of insecticide used between the areas. Bioassays with larvae and adult were performed according to the World Health Organization methods. The results showed higher resistance levels to organophosphates group in populations from the Northeast region where this group was used for both larvae and adult control than in S?o Paulo where organophosphates were used for larvae and pyretroids for adult control. Resistance to pyretroids in adults was widespread in S?o Paulo after ten years of use of cypermethrin while in vector populations from the Northeast region it was punctual. The difference in resistance profile between the areas is in accordance to the group of insecticide used.     

Pyrethroid resistance in Aedes aegypti and Aedes albopictus from Port‐au‐Prince,Haiti

In Port‐au‐Prince, Haiti, the status of insecticide resistance has not recently been evaluated for Aedes aegypti (L) and Aedes albopictus (Skuse) populations. No prophylactics exist for dengue, so prevention is only through vector control methods. An earthquake occurred in Haiti on January 12, 2010, with a magnitude of 7.0 Mw that devastated the area. Dengue became a major concern for the humanitarian relief workers that entered the country. Bottle bioassays were conducted in the field on adult mosquitoes reared from larvae collected from the grounds of the U.S. Embassy and from an adjacent neighborhood in eastern Port‐au‐Prince, Haiti. At the CDC, Fort Collins, CO, bioassays, molecular, and biochemical assays were performed on mosquitoes reared from field‐collected eggs. A small percentage of the population was able to survive the diagnostic dose in bioassays run in Haiti. Mosquitoes tested at the CDC demonstrated no phenotypic resistance. A variety of factors could be responsible for the discrepancies between the field and lab data, but temperature and larval nutrition are probably most important. Knowledge of localized resistance and underlying mechanisms helps in making rational decisions in selection of appropriate and effective insecticides in the event of a dengue outbreak.     

Elevated activity of an Epsilon class glutathione transferase confers DDT resistance in the dengue vector, Aedes aegypti

Glutathione transferases (GSTs) play a central role in the detoxification of xenobiotics such as insecticides and elevated GST expression is an important mechanism of insecticide resistance. In the mosquito, Anopheles gambiae, increased expression of an Epsilon class GST, GSTE2, confers resistance to DDT. We have identified eight GST genes in the dengue vector, Aedes aegypti. Four of these belong to the insect specific GST classes Delta and Epsilon and three are from the more ubiquitously distributed Theta and Sigma classes. The expression levels of the two Epsilon genes, a Theta GST and a previously identified Ae. aegypti GST [Grant and Hammock, 1992. Molecular and General Genetics 234, 169-176] were established for an insecticide susceptible and a resistant strain. We show that the putative ortholog of GSTe2 in Ae. aegypti (AaGSTe2) is over expressed in mosquitoes that are resistant to the insecticides DDT and permethrin. Characterisation of recombinant AaGSTE2-2 confirmed the role of this enzyme in DDT metabolism. In addition, unlike its Anopheles ortholog, AaGSTE2-2 also exhibited glutathione peroxidase activity.     

Effect of Moringa oleifera flower extract on larval trypsin and acetylcholinesterase activities in Aedes aegypti

Aedes aegypti control is crucial to reducing dengue fever. Aedes aegypti larvae have developed resistance to organophosporous insecticides and the use of natural larvicides may help manage larval resistance by increasing elements in insecticide rotation programs. Here, we report on larvicidal activity of Moringa oleifera flower extract against A. aegypti L(1), L(2), L(3), and L(4) as well as the effect of flower extract on gut trypsin and whole-larval acetylcholinesterase from L(4.) In addition, the heated flower extract was investigated for larvicidal activity against L(4) and effect on larval gut trypsin. Moringa oleifera flower extract contains a proteinaceous trypsin inhibitor (M. oleifera flower trypsin inhibitor, MoFTI), triterpene (β-amyrin), sterol (β-sitosterol) as well as flavonoids (kaempferol and quercetin). Larvicidal activity was detected against L(2), L(3), and L(4) (LC(50) of 1.72%, 1.67%, and 0.92%, respectively). Flower extract inhibited L(4) gut trypsin (MoFTI K(i) = 0.6 nM) and did not affect acetylcholinesterase activity. In vivo assay showed that gut trypsin activity from L(4) treated with M. oleifera flower extract decreased over time (0-1,440 min) and was strongly inhibited (98.6%) after 310 min incubation; acetylcholinesterase activity was not affected. Thermal treatment resulted in a loss of trypsin inhibitor and larvicidal activities, supporting the hypothesis that flower extract contains a proteinaceous trypsin inhibitor that may be responsible for the deleterious effects on larval mortality.     

Estimation of Aedes aegypti (Diptera: Culicidae) population size and adult male survival in an urban area in Panama

Traditional mosquito control strategies rely heavily on the use of chemical insecticides. However, concerns about the efficiency of traditional control methods, environmental impact and emerging pesticide resistance have highlighted the necessity for developing innovative tools for mosquito control. Some novel strategies, including release of insects carrying a dominant lethal gene (RIDL^®), rely on the sustained release of modified male mosquitoes and therefore benefit from a thorough understanding of the biology of the male of the species. In this report we present the results of a mark-release-recapture study aimed at: (i) establishing the survival in the field of laboratory-reared, wild-type male Aedes aegypti and (b) estimating the size of the local adult Ae. aegypti population. The study took place in Panama, a country where recent increases in the incidence and severity of dengue cases have prompted health authorities to evaluate alternative strategies for vector control. Results suggest a life expectancy of 2.3 days for released male mosquitoes (confidence interval: 1.78-2.86). Overall, the male mosquito population was estimated at 58 males/ha (range 12-81 males/ha), which can be extrapolated to an average of 0.64 pupae/person for the study area. The practical implications of these results are discussed.     

Field evaluation of a lethal ovitrap against dengue vectors in Brazil

Field evaluation of a "lethal ovitrap" (LO) to control dengue vector Aedes mosquitoes (Diptera: Culicidae), was undertaken in two Brazilian municipalities, Areia Branca and Nilopolis, in the State of Rio de Janeiro. The LO is designed to kill Aedes via an insecticide-treated ovistrip (impregnated with deltamethrin). In each municipality, the intervention was applied to a group of 30 houses (10 LOs/house) and compared to 30 houses without LOs in the same neighbourhood. Five LOs were put outside and five LOs inside each treated house. Three methods of monitoring Aedes density were employed: (i) percentage of containers positive for larvae and/or pupae; (ii) total pupae/house; (iii) total adult females/house collected by aspirator indoors. Weekly mosquito surveys began during the month before LO placement, by sampling from different groups of 10 houses/week for 3 weeks pre-intervention (i.e. 30 houses/month) and for 3 months post-intervention in both treated and untreated areas. Prior to LO placement at the end of February 2001, Aedes aegypti (L) densities were similar among houses scheduled for LO treatment and comparison (untreated control) at each municipality. Very few Ae. albopictus (Skuse) were found and this species was excluded from the assessment. Post-intervention densities of Ae. aegypti were significantly reduced for most comparators (P < 0.01), as shown by fewer positive containers (4-5 vs. 10-18) and pupae/house (0.3-0.7 vs. 8-10) at LO-treated vs. untreated houses, 3 months post-treatment at both municipalities. Numbers of adult Ae. aegypti females indoors were consistently reduced in LO-treated houses at Areia Branca (3.6 vs. 6.8/house 3 months post-intervention) but not at Niloplis (approximately 3/house, attributed to immigration). These results demonstrate sustained impact of LOs on dengue vector population densities in housing conditions of Brazilian municipalities.     

Insecticide resistance in dengue vectors from hotspots in Selangor,Malaysia

BackgroundIn Malaysia, dengue remains a top priority disease and usage of insecticides is the main method for dengue vector control. Limited baseline insecticide resistance data in dengue hotspots has prompted us to conduct this study. The present study reports the use of a map on the insecticide susceptibility status of Aedes aegypti and Aedes albopictus to provide a quick visualization and overview of the distribution of insecticide resistance.Method and resultsThe insecticide resistance status of Aedes populations collected from 24 dengue hotspot areas from the period of December 2018 until June 2019 was proactively monitored using the World Health Organization standard protocol for adult and larval susceptibility testing was conducted, together with elucidation of the mechanisms involved in observed resistance. For resistance monitoring, susceptibility to three adulticides (permethrin, deltamethrin, and malathion) was tested, as well as susceptibility to the larvicide, temephos. Data showed significant resistance to both deltamethrin and permethrin (pyrethroid insecticides), and to malathion (organophosphate insecticide) in all sampled Aedes aegypti populations, while variable resistance patterns were found in the sampled Aedes albopictus populations. Temephos resistance was observed when larvae were tested using the diagnostic dosage of 0.012mg/L but not at the operational dosage of 1mg/L for both species.ConclusionThe present study highlights evidence of a potential threat to the effectiveness of insecticides currently used in dengue vector control, and the urgent requirement for insecticide resistance management to be integrated into the National Dengue Control Program.