蚊媒传染病的遗传控制和共生控制

蚊虫是疟疾和登革热等多种疾病的传播媒介,媒介控制是阻断虫媒传染病的重要措施。当前对媒介的控制主要依赖于化学杀虫剂,但蚊虫已对杀虫剂产生了普遍抗性,加上疟原虫等耐药性问题的出现和抗疟疫苗的缺乏,急需发展新的方法和策略用于蚊媒传染病的防控。蚊子中肠是疟原虫等病原体在蚊虫体内发育的最大屏障,是阻断疾病传播的理想靶点。基于转基因蚊的遗传控制和转基因共生菌的共生控制是降低媒介效能和阻断疾病传播的两个有前景的新策略。遗传控制是直接以媒介昆虫作为遗传操作对象,通过表达抗病效应分子来阻断疾病的传播;转基因共生菌防治则是以共生微生物作为遗传改造的对象,在宿主体内表达抗病效应分子以达到阻断疾病传播的目的。本文对这些新防治方法的现状及应用进展进行综述,并讨论遗传控制和共生控制在蚊媒传染病防治的实际应用中所面临的问题。     

害虫遗传不育技术中致死基因的研究与应用

基于遗传修饰手段的昆虫不育技术(SIT)作为一类物种特异、环境友好、科学高效的新兴策略,在害虫防治中具有广阔的应用前景。释放携带显性致死基因昆虫的技术(RIDL)是改进传统SIT的重要手段之一,主要包括四环素调控系统、特异性启动子、性别特异剪接系统和特异性致死基因等重要元件,其中根据不同昆虫的特点选择合适的特异性致死基因对于构建遗传不育品系至关重要。这些致死基因或受到阻遏调控系统的控制、或特异的在雌虫中表达、亦或直接作用于X染色体,导致后代在特定发育阶段或特定性别中条件致死。本文综述了RHG家族(reapr、hid、grim、michelob_x)细胞凋亡基因、转录激活因子t TA及Nipp1Dm、归巢内切酶基因等在害虫遗传不育技术中的研究和应用,讨论了特定致死基因的效应机理和应用特点,并对其可能的发展方向进行了展望。由于不同效应基因的致死作用和调控机理尚未完全明晰,因此深入研究特异致死基因的凋亡机制和在不同物种中的兼容作用,将为害虫遗传防控提供更多的研究思路和手段。     

苹果蠹蛾的综合防控和遗传控制研究进展

苹果蠹蛾是仁果类水果的重要检疫害虫,在世界各地造成了巨大的经济损失。目前对其化学防治、化学生态调控、病毒等防治方法研究较多,但仍不能满足防控该害虫的需要,对新型防控技术的需求日益增强。不育昆虫释放技术(SIT)是一种可控制甚至根除靶标害虫的环境友好型防控技术,但传统SIT技术存在一定的局限性,如较难区分性别与筛选雌雄虫、辐射不育昆虫的交配竞争力和适合度降低等问题,这些缺陷随着昆虫遗传修饰技术的发展将得以解决,并将在害虫防控进程中起到积极作用。本文综述了苹果蠹蛾主要防控技术研究现状,介绍了通过遗传修饰技术改善SIT的技术策略,并综合分析了我国开展苹果蠹蛾遗传修饰研究情况和将其应用在苹果蠹蛾防控体系中的可行性及优势。     

蚊虫微生物群系及其在蚊媒疾病防控中的研究进展

蚊虫体内外栖居着大量的微生物,构成多样且动态的微生物群系(microbiome).蚊虫微生物群系(mosquito microbiome)在蚊虫的营养、代谢、免疫等诸多生理功能上发挥着重要作用,是蚊虫生长发育不可或缺的重要组成部分.同时,蚊虫微生物群系显著影响蚊媒病原体感染和传播,在蚊媒疾病防控中具有重要的应用价值.因此,全面了解蚊虫生理以及蚊媒疾病传播不能忽视微生物群系的关键作用.本文系统概述了蚊虫微生物群系的组成及其影响因素、微生物群系对宿主生理功能的影响与互作机制,以及基于蚊虫微生物群系的蚊媒疾病阻断控制等方面的研究进展,并对利用微生物群系防控蚊媒传染病的应用前景提出展望.     

生物策略在控制伊蚊传播疾病的应用

RNAi技术作为高效、特异性的阻断技术,已经在低等原核生物、植物、动物、真菌等的研究和农作物病虫害防治、人类蚊媒传染病的控制等方面得到了广泛的应用。近年来全球每年有约一半的人受到蚊媒传播疾病的威胁,蚊媒传病严重危害全世界人类的生命健康,成为巨大的公共卫生问题。其中,伊蚊在传播疾病方面扮演着十分重要的角色,包括目前最重要的流行性病,如登革热、寨卡病毒病、黄热病、流行性乙型脑炎、基孔肯雅病等都是由伊蚊传播的疾病。因此,非常急需一种有效的防控蚊虫的手段来控制疾病的传播。生物防控策略具有环境和生态友好的特征,为近年研究的主流。本研究从近十年来伊蚊生物防控的研究成果,着重综述了生物防控策略的有效性、可行性和经济性,对本领域研究进展和未来的发展方向进行了总结和阐述。     

Wolbachia在蚊虫中的防控应用研究进展

蚊媒传播的疾病在世界范围内造成巨大的经济损失和健康负担。传统化学杀虫剂的不规范和过度使用导致抗药性在蚊虫种群的扩散,加剧了蚊媒传播疾病的发生,因此,研发经济安全的新型蚊媒防控方法至关重要。Wolbachia是一种专性感染节肢动物的细胞内共生菌,可以操纵蚊虫的生殖能力并具有病原体阻断作用。释放Wolbachia感染的蚊虫品系来抑制或修饰蚊虫种群的防控策略具有巨大的发展潜力。本文主要介绍了Wolbachia诱导的胞质不相容作用和Wolbachia介导的病原体阻断作用,并阐述了以这两种技术为基础的蚊虫防控策略的研究进展,包括种群抑制和种群修饰策略。     

害虫遗传防控技术的研究与应用

遗传防控技术作为传统有害生物防控方式的替代策略,自20世纪50年代以来在世界各国受到广泛的关注,被认为是一类有利于人类健康、食品安全和农业可持续发展的害虫种群防控技术,同时也为日益严峻的生物入侵问题提供了解决方案.本文综述了不育昆虫技术(sterile insect technique, SIT)、释放携带显性致死基因昆虫技术(release of insects carrying a dominant lethal, RIDL)和基因驱动技术(gene drive)的基本原理及部分应用案例,比较了不同技术的优势与限制因素,并介绍了相关研究中常见的几种基因整合策略,以期进一步改进害虫综合治理的基础研究和技术研发.     

遗传控制技术在实蝇类害虫中的研究进展

实蝇类害虫严重危害多种水果和蔬菜,是世界果蔬产业最重要的害虫类群之一,严重影响了发生地的果蔬生产和出口贸易活动。昆虫不育技术(SIT)是一种物种特异和环境友好型防治措施,在多种实蝇类害虫的防治、阻截和根除中起到了不可替代的重要作用。通过分子生物学技术对昆虫的基因组进行遗传修饰,可对SIT进行改进,提高其防控效果并扩大应用的物种范围,近年来相关方面的研究已取得重要进展,成为害虫遗传控制的研究热点。本文阐述了通过受四环素调控的tet-off基因表达系统来实现昆虫"不育"的基本原理和在果蝇及其他几种主要实蝇类害虫中建立的不同类型的遗传控制体系,以及类似体系在其他农业昆虫中的应用情况。简要介绍了在橘小实蝇遗传控制技术体系构建方面的工作进展,并对该技术的在害虫综合治理(IPM)尤其是实蝇类害虫防治中的应用前景进行了讨论和展望。     

RNAi在抗蚊媒病毒感染中的研究进展

蚊媒病因具有较高的发病率和传播率使其成为全球关注的重要公共卫生问题。蚊虫作为蚊媒病的传播媒介,研究其与蚊媒病毒两者之间的相互作用机制将有助于蚊媒病的防控。蚊虫抵御蚊媒病毒的先天免疫降低和病毒成功逃避蚊虫免疫屏障为病毒在蚊虫体内的持续感染和蚊媒病的暴发流行造成了潜在风险。RNA干扰(RNA interference, RNAi)途径作为蚊虫体内强大的抗病毒防御屏障,通过产生多种小RNA降解病毒RNA,从而达到抑制病毒复制和传播的目的。本文对小干扰RNA (small interfering RNA, si RNA)、微小RNA (micro RNA,mi RNA)、Piwi蛋白相作用RNA (Piwi-interacting RNA, pi RNA)等3种小分子RNA在蚊虫体内发挥抗蚊媒病毒感染的先天免疫机制的相关研究进行了综述,以期为蚊媒病的防控提供理论参考。     

昆虫内共生菌沃尔巴克氏体抗病毒研究进展

植物病毒病是危害我国蔬菜生产的第一大病害,而烟粉虱Bemisia tabaci Gennadius、蓟马和蚜虫等小型昆虫是蔬菜病毒病的主要传播媒介.虫传病毒病害的防控策略复杂且难度大,目前生产上主要依赖化学农药防治介体昆虫,预防与控制蔬菜病毒病.种植户化学杀虫药剂的不合理使用、甚至滥用,导致媒介昆虫抗药性、杀虫剂污染与残留等问题,严重威胁农产品安全与生态安全.发展高效控制媒介昆虫及其传播重大病毒病害防控技术是保障食品安全、蔬菜产业健康可持续发展的迫切需求.内共生菌沃尔巴克氏体Wolbachia是节肢动物体内广泛存在并经卵传播的革兰氏阴性胞内次生共生细菌,约65％的昆虫天然携带沃尔巴克氏体.沃尔巴克氏体对宿主昆虫具有胞质不亲和、产雌孤雌生殖、雌性化或者杀雄作用等生殖调控作用,同时抑制虫媒病毒病在昆虫体内的复制和传播.基于沃尔巴克氏体的蚊媒及蚊媒病毒病控制研究与应用取得了重大进展,现就沃尔巴克氏体抗病毒及其应用的最新进展和未来发展进行综述,为发展蔬菜害虫及其传播病毒病防控技术提供新思路.     

A model framework to estimate impact and cost of genetics-based sterile insect methods for dengue vector control

Vector-borne diseases impose enormous health and economic burdens and additional methods to control vector populations are clearly needed. The Sterile Insect Technique (SIT) has been successful against agricultural pests, but is not in large-scale use for suppressing or eliminating mosquito populations. Genetic RIDL technology (Release of Insects carrying a Dominant Lethal) is a proposed modification that involves releasing insects that are homozygous for a repressible dominant lethal genetic construct rather than being sterilized by irradiation, and could potentially overcome some technical difficulties with the conventional SIT technology. Using the arboviral disease dengue as an example, we combine vector population dynamics and epidemiological models to explore the effect of a program of RIDL releases on disease transmission. We use these to derive a preliminary estimate of the potential cost-effectiveness of vector control by applying estimates of the costs of SIT. We predict that this genetic control strategy could eliminate dengue rapidly from a human community, and at lower expense (approximately US$ 2~30 per case averted) than the direct and indirect costs of disease (mean US$ 86-190 per case of dengue). The theoretical framework has wider potential use; by appropriately adapting or replacing each component of the framework (entomological, epidemiological, vector control bio-economics and health economics), it could be applied to other vector-borne diseases or vector control strategies and extended to include other health interventions.     

Genetic elimination of field-cage populations of Mediterranean fruit flies

The Mediterranean fruit fly (medfly, Ceratitis capitata Wiedemann) is a pest of over 300 fruits, vegetables and nuts. The sterile insect technique (SIT) is a control measure used to reduce the reproductive potential of populations through the mass release of sterilized male insects that mate with wild females. However, SIT flies can display poor field performance, due to the effects of mass-rearing and of the irradiation process used for sterilization. The development of female-lethal RIDL (release of insects carrying a dominant lethal) strains for medfly can overcome many of the problems of SIT associated with irradiation. Here, we present life-history characterizations for two medfly RIDL strains, OX3864A and OX3647Q. Our results show (i) full functionality of RIDL, (ii) equivalency of RIDL and wild-type strains for life-history characteristics, and (iii) a high level of sexual competitiveness against both wild-type and wild-derived males. We also present the first proof-of-principle experiment on the use of RIDL to eliminate medfly populations. Weekly releases of OX3864A males into stable populations of wild-type medfly caused a successive decline in numbers, leading to eradication. The results show that genetic control can provide an effective alternative to SIT for the control of pest insects.     

害虫遗传防治的研究历史与现状

作为防治或根除重大害虫最为有效的手段之一,害虫遗传防治在世界范围内被广泛采用并取得了巨大成功。本文综述了不育昆虫技术、雌性致死系统和昆虫显性致死技术等经典害虫遗传防治策略的发展历史、技术特点和应用情况。近年来,许多新的分子生物手段被不断提出并整合到害虫遗传防治策略中,包括归巢核酸内切酶基因、锌指核酸酶、转录激活因子样效应因子核酸酶、CRISPR/Cas9系统、Medea元件、Killer-Rescue系统、Wolbachia-细胞质不亲和性系统等。基于这些新的工具手段,许多国家已经在不同程度上启动了下一代害虫遗传防治项目。而我国在该领域的研究相对薄弱,需要在借鉴国外成功经验的同时,进一步加强害虫遗传防治的基础和应用研究,从而实现本地有害生物的可持续治理和外来入侵生物的有效狙击,确保我国未来的粮食和生态安全。     

High-throughput sorting of mosquito larvae for laboratory studies and for future vector control interventions

ABSTRACT: BACKGROUND: Mosquito transgenesis offers new promises for the genetic control of vector-borne infectious diseases such as malaria and dengue fever. Genetic control strategies require the release of large number of male mosquitoes into field populations, whether they are based on the use of sterile males (sterile insect technique, SIT) or on introducing genetic traits conferring refractoriness to disease transmission (population replacement). However, the current absence of high-throughput techniques for sorting different mosquito populations impairs the application of these control measures. METHODS: A method was developed to generate large mosquito populations of the desired sex and genotype. This method combines flow cytometry and the use of Anopheles gambiae transgenic lines that differentially express fluorescent markers in males and females. RESULTS: Fluorescence-assisted sorting allowed single-step isolation of homozygous transgenic mosquitoes from a mixed population. This method was also used to select wild-type males only with high efficiency and accuracy, a highly desirable tool for genetic control strategies where the release of transgenic individuals may be problematic. Importantly, sorted males showed normal mating ability compared to their unsorted brothers. CONCLUSIONS: The developed method will greatly facilitate both laboratory studies of mosquito vectorial capacity requiring high-throughput approaches and future field interventions in the fight against infectious disease vectors.     

Pair of lice lost or parasites regained: the evolutionary history of anthropoid primate lice

Background

Reduction or elimination of vector populations will tend to reduce or eliminate transmission of vector-borne diseases. One potential method for environmentally-friendly, species-specific population control is the Sterile Insect Technique (SIT). SIT has not been widely used against insect disease vectors such as mosquitoes, in part because of various practical difficulties in rearing, sterilization and distribution. Additionally, vector populations with strong density-dependent effects will tend to be resistant to SIT-based control as the population-reducing effect of induced sterility will tend to be offset by reduced density-dependent mortality.

Results

We investigated by mathematical modeling the effect of manipulating the stage of development at which death occurs (lethal phase) in an SIT program against a density-dependence-limited insect population. We found late-acting lethality to be considerably more effective than early-acting lethality. No such strains of a vector insect have been described, so as a proof-of-principle we constructed a strain of the principal vector of the dengue and yellow fever viruses, Aedes (Stegomyia) aegypti, with the necessary properties of dominant, repressible, highly penetrant, late-acting lethality.

Conclusion

Conventional SIT induces early-acting (embryonic) lethality, but genetic methods potentially allow the lethal phase to be tailored to the program. For insects with strong density-dependence, we show that lethality after the density-dependent phase would be a considerable improvement over conventional methods. For density-dependent parameters estimated from field data for Aedes aegypti, the critical release ratio for population elimination is modeled to be 27% to 540% greater for early-acting rather than late-acting lethality. Our success in developing a mosquito strain with the key features that the modeling indicated were desirable demonstrates the feasibility of this approach for improved SIT for disease control.     

Heritable strategies for controlling insect vectors of disease

Mosquito-borne diseases are causing a substantial burden of mortality, morbidity and economic loss in many parts of the world, despite current control efforts, and new complementary approaches to controlling these diseases are needed. One promising class of new interventions under development involves the heritable modification of the mosquito by insertion of novel genes into the nucleus or of Wolbachia endosymbionts into the cytoplasm. Once released into a target population, these modifications can act to reduce one or more components of the mosquito population''s vectorial capacity (e.g. the number of female mosquitoes, their longevity or their ability to support development and transmission of the pathogen). Some of the modifications under development are designed to be self-limiting, in that they will tend to disappear over time in the absence of recurrent releases (and hence are similar to the sterile insect technique, SIT), whereas other modifications are designed to be self-sustaining, spreading through populations even after releases stop (and hence are similar to traditional biological control). Several successful field trials have now been performed with Aedes mosquitoes, and such trials are helping to define the appropriate developmental pathway for this new class of intervention.     

Mosquito management in the face of natural selection

The sterile insect technique (SIT) is an appealing method for managing mosquito populations while avoiding the environmental and social costs associated with more traditional control strategies like insecticide application. Success of SIT, however, hinges on sterile males being able to compete for females. As a result, heavy and/or continued use of SIT could potentially diminish its efficacy if prolonged treatments result in selection for female preference against sterile males. In this paper we extend a general differential equation model of mosquito dynamics to consider the role of female choosiness in determining the long-term usefulness of SIT as a management option. We then apply optimal control theory to our model and show how natural selection for female choosiness fundamentally alters management strategies. Our study calls into question the benefits associated with developing SIT as a management strategy, and suggests that effort should be spent studying female mate choice in order to determine its relative importance and how likely it is to impact SIT treatment goals.     

The first releases of transgenic mosquitoes: an argument for the sterile insect technique

Potential applications for reducing transmission of mosquito-borne diseases by releasing genetically modified mosquitoes have been proposed, and mosquitoes are being created with such an application in mind in several laboratories. The use of the sterile insect technique (SIT) provides a safe programme in which production, release and mating competitiveness questions related to mass-reared genetically modified mosquitoes could be answered. It also provides a reversible effect that would be difficult to accomplish with gene introgression approaches. Could new technologies, including recombinant DNA techniques, have improved the success of previous mosquito releases? Criteria for an acceptable transgenic sterile mosquito are described, and the characteristics of radiation-induced sterility are compared with that of current transgenic approaches. We argue that SIT using transgenic material would provide an essentially safe and efficacious foundation for other possible approaches that are more ambitious.     

Developing arbovirus resistance in mosquitoes

Diseases caused by arthropod-borne viruses are increasingly significant public health problems, and novel methods are needed to control pathogen transmission. The hypothesis underlying the research described here is that genetic manipulation of Aedes aegypti mosquitoes can profoundly and permanently reduce their competence to transmit dengue viruses to human hosts. Recent key findings now allow us to test the genetic control hypothesis. We have identified viral genome-derived RNA segments that can be expressed in mosquito midguts and salivary glands to ablate homologous virus replication and transmission. We have demonstrated that both transient and heritable expression of virus-derived effector RNAs in cultured mosquito cells can silence virus replication, and have characterized the mechanism of RNA-mediated resistance. We are now developing virus-resistant mosquito lines by transformation with transposable elements that express effector RNAs from mosquito-active promoters.