Khan Jehangir, Adil Muhammad, Junyan Zhang, Zhang Dongjing, Deng Yidong, Lv Zhiyue, Chen Tao
Hainan General Hospital, Hainan Affiliated Hospital of Hainan Medical University, Haikou, 570100, Hainan, China.
Laboratory of Tropical Veterinary Medicine and Vector Biology, School of Life Sciences, Hainan University, Haikou, 570228, China.
Parasit Vectors. 2025 Jun 1;18(1):201. doi: 10.1186/s13071-025-06826-8.
Understanding the intrinsic factors that influence mosquito vector competence (VC) to pathogens is crucial for assessing the risk of disease transmission in both rural and urban environments. We assessed the VC of Aedes albopictus mosquitoes from urban (dengue-endemic) and rural (dengue-free) areas in Guangzhou, China, for dengue virus-2 (DENV-2), while also examining intrinsic factors such as Wolbachia and immune-related gene expression influencing VC.
Adult females of rural, urban, and laboratory (control) populations of Ae. albopictus were orally exposed to a freshly prepared suspension of the DENV-2 New Guinea C strain (GenBank: AF038403.1), with a final titer of 1 × 10 plaque-forming units (PFU)/ml, for a period of 60 min. Three different bioassays (B1-B3) were conducted on 60 mosquitoes per population: B1 at 7 days post-exposure (dpe) to assess viral infection in the mosquito midgut, and B2 and B3 at 14 dpe to evaluate viral dissemination in the carcass and transmission via saliva. The mosquito samples were processed for total RNA and DNA extraction. RNA was subsequently analyzed by quantitative reverse transcription polymerase chain reaction (qRT-PCR) to quantify viral load and measure the expression of immune-related genes, while DNA was assessed via quantitative PCR (qPCR) to determine Wolbachia density (wAlbA and wAlbB) and the rps6 gene.
At 7 dpe, virus proliferation in rural mosquitoes was similar to that in urban mosquitoes (P = 0.10). By 14 dpe, rural mosquitoes showed a significantly lower status of virus dissemination (P < 0.04) and transmission (P < 0.012). Wolbachia (-0.12 < r < -0.92) and immune effectors (-0.025 < r < -0.568) were negatively correlated with DENV in all mosquitoes, with more negative values indicating a stronger inverse relationship. The wAlbA and wAlbB strains exhibited similar densities across all the mosquito populations, with wAlbB revealing a slightly greater abundance in rural mosquitoes, although the difference was not significant. Elevated Relish 2 (Rel2), defensin A (DefA), and the signal transducers and activators of transcription (STAT) levels indicate activation of the Toll and JAK-STAT pathways, contributing to resistance against DENV replication and reduced VC in rural mosquitoes.
This study indicates that rural Ae. albopictus mosquitoes may possess intrinsic barriers limiting their VC for DENV-2, offering valuable preliminary insights into VC across geographically distinct populations. However, further research across a broader range of urban and rural locations is needed to validate these findings and better understand the local factors influencing VC. Such insights are vital for public health, as they can help prioritize locations for dengue surveillance and effective vector control. Future studies should investigate the roles of intestinal microflora and immune pathways across diverse mosquito populations to better understand these dynamics and improve our knowledge of mosquito-borne disease transmission in different environments.
了解影响蚊媒对病原体传播能力(VC)的内在因素对于评估农村和城市环境中疾病传播风险至关重要。我们评估了中国广州城市(登革热流行)和农村(无登革热)地区的白纹伊蚊对登革病毒2型(DENV - 2)的传播能力,同时还研究了诸如沃尔巴克氏体和影响传播能力的免疫相关基因表达等内在因素。
将农村、城市和实验室(对照)种群的成年雌性白纹伊蚊口服暴露于新鲜制备的DENV - 2新几内亚C株(GenBank:AF038403.1)悬浮液中,终浓度为1×10斑块形成单位(PFU)/毫升,持续60分钟。对每个种群的60只蚊子进行三种不同的生物测定(B1 - B3):B1在暴露后7天(dpe)进行,以评估蚊子中肠的病毒感染情况;B2和B3在暴露后14天进行,以评估病毒在虫体中的传播以及通过唾液的传播情况。对蚊子样本进行总RNA和DNA提取。随后通过定量逆转录聚合酶链反应(qRT - PCR)分析RNA,以量化病毒载量并测量免疫相关基因的表达,而通过定量PCR(qPCR)评估DNA,以确定沃尔巴克氏体密度(wAlbA和wAlbB)以及rps6基因。
在暴露后7天,农村蚊子中的病毒增殖与城市蚊子相似(P = 0.10)。到暴露后14天,农村蚊子的病毒传播(P < 0.04)和传播能力(P < 0.012)显著较低。在所有蚊子中,沃尔巴克氏体(-0.12 < r < -0.92)和免疫效应因子(-0.025 < r < -0.568)与DENV呈负相关,负值越大表明反向关系越强。wAlbA和wAlbB菌株在所有蚊子种群中的密度相似,wAlbB在农村蚊子中的丰度略高,尽管差异不显著。Relish 2(Rel2)、防御素A(DefA)以及信号转导和转录激活因子(STAT)水平升高表明Toll和JAK - STAT途径被激活,这有助于农村蚊子抵抗DENV复制并降低传播能力。
本研究表明农村白纹伊蚊可能具有限制其对DENV - 2传播能力的内在障碍,为不同地理种群的传播能力提供了有价值的初步见解。然而,需要在更广泛的城乡地区进行进一步研究以验证这些发现并更好地了解影响传播能力的当地因素。这些见解对公共卫生至关重要,因为它们有助于确定登革热监测和有效病媒控制的重点地区。未来的研究应调查不同蚊子种群中肠道微生物群和免疫途径的作用,以更好地理解这些动态并增进我们对不同环境中蚊媒疾病传播的认识。