Multiple Salivary Proteins from Aedes aegypti Mosquito Bind to the Zika Virus Envelope Protein.

Aedes aegypti mosquitoes are important vectors of several debilitating and deadly arthropod-borne (arbo) viruses, including Yellow Fever virus, Dengue virus, West Nile virus and Zika virus (ZIKV). Arbovirus transmission occurs when an infected mosquito probes the host's skin in search of a blood meal. Salivary proteins from mosquitoes help to acquire blood and have also been shown to enhance pathogen transmission in vivo and in vitro. Here, we evaluated the interaction of mosquito salivary proteins with ZIKV by surface plasmon resonance and enzyme-linked immunosorbent assay. We found that three salivary proteins AAEL000793, AAEL007420, and AAEL006347 bind to the envelope protein of ZIKV with nanomolar affinities. Similar results were obtained using virus-like particles in binding assays. These interactions have no effect on viral replication in cultured endothelial cells and keratinocytes. Additionally, we found detectable antibody levels in ZIKV and DENV serum samples against the recombinant proteins that interact with ZIKV. These results highlight complex interactions between viruses, salivary proteins and antibodies that could be present during viral transmissions.

Salivary factor LTRIN from Aedes aegypti facilitates the transmission of Zika virus by interfering with the lymphotoxin-ß receptor.

Pathogens have co-evolved with mosquitoes to optimize transmission to hosts. Mosquito salivary-gland extract is known to modulate host immune responses and facilitate pathogen transmission, but the underlying molecular mechanisms of this have remained unknown. In this study, we identified and characterized a prominent 15-kilodalton protein, LTRIN, obtained from the salivary glands of the mosquito Aedes aegypti. LTRIN expression was upregulated in blood-fed mosquitoes, and LTRIN facilitated the transmission of Zika virus (ZIKV) and exacerbated its pathogenicity by interfering with signaling through the lymphotoxin-ß receptor (LTßR). Mechanically, LTRIN bound to LTßR and 'preferentially' inhibited signaling via the transcription factor NF-κB and the production of inflammatory cytokines by interfering with the dimerization of LTßR during infection with ZIKV. Furthermore, treatment with antibody to LTRIN inhibited mosquito-mediated infection with ZIKV, and abolishing LTßR potentiated the infectivity of ZIKV both in vitro and in vivo. This study provides deeper insight into the transmission of mosquito-borne diseases in nature and supports the therapeutic potential of inhibiting the action of LTRIN to disrupt ZIKV transmission.

High efficacy of (Z)-γ-bisabolene from the essential oil of Galinsoga parviflora (Asteraceae) as larvicide and oviposition deterrent against six mosquito vectors.

The eco-friendly management of mosquitoes with novel and effective larvicides and oviposition deterrents is a crucial challenge to prevent outbreaks of mosquito-borne diseases. However, most of the herbal formulations tested in these years showed LC50 values higher of 40 ppm, and significant oviposition deterrent activity only when tested at relatively higher doses (> 50 µg/ml). Herein, we studied the chemical composition of the Galinsoga parviflora essential oil (EO). This plant is an annual herb native to South America naturalized all over the world. We tested the EO larvicidal and oviposition deterrent action on 6 mosquito species. Totally 37 compounds were identified in the EO of G. parviflora by GC and GC-MS analyses. The major constituent was (Z)-γ-bisabolene (38.9%). The G. parviflora EO and (Z)-γ-bisabolene showed acute toxicity on An. stephensi (LC50 = 31.04 and 2.04 µg/ml, respectively), Ae. aegypti (LC50 = 34.22 and 2.26 µg/ml, respectively), Cx. quinquefasciatus (LC50 = 37.10 and 2.47 µg/ml, respectively), An. subpictus (LC50 = 40.97 and 4.09 µg/ml, respectively), Ae. albopictus (LC50 = 45.55 and 4.50 µg/ml, respectively) and Cx. tritaeniorhynchus (LC50 = 49.56 and 4.87 µg/ml, respectively) larvae. Furthermore, the oviposition deterrent potential of the G. parviflora EO and (Z)-γ-bisabolene was studied on six mosquito vectors, showing that 25 µg/ml of (Z)-γ-bisabolene led to an Oviposition Activity Index lower of - 0.79 in all tested mosquito vectors. Overall, all larvicidal LC50 values estimated for (Z)-γ-bisabolene were lower than 5 µg/ml. This result far encompasses current evidences of toxicity reported for the large majority of botanical products currently tested against mosquito young instars, allowing us to propose this compound as an highly effective mosquito larvicide and oviposition deterrent.

Fabrication of highly effective mosquito nanolarvicides using an Asian plant of ethno-pharmacological interest, Priyangu (Aglaia elaeagnoidea): toxicity on non-target mosquito natural enemies.

Mosquitoes threaten the lives of humans, livestock, pets and wildlife around the globe, due to their ability to vector devastating diseases. Aglaia elaeagnoidea, commonly known as Priyangu, is widely employed in Asian traditional medicine and pest control. Medicinal activities include anti-inflammatory, analgesic, anticancer, and anesthetic actions. Flavaglines, six cyclopenta[b]benzofurans, a cyclopenta[bc]benzopyran, a benzo[b]oxepine, and an aromatic butyrolactone showed antifungal properties, and aglaroxin A and rocaglamide were effective to control moth pests. Here, we determined the larvicidal action of A. elaeagnoidea leaf aqueous extract. Furthermore, we focused on Priyangu-mediated synthesis of Ag nanoparticles toxic to Culex quinquefasciatus, Aedes aegypti and Anopheles stephensi. The plant extract and the nanolarvicide were tested on three mosquito vectors, following the WHO protocol, as well as on three non-target mosquito predators. Priyangu-synthesized Ag nanoparticles were characterized by spectroscopic (UV, FTIR, XRD, and EDX) and microscopic (AFM, SEM, and TEM) analyses. Priyangu extract toxicity was moderate on Cx. quinquefasciatus (LC50 246.43; LC90 462.09 µg/mL), Ae. aegypti (LC50 229.79; LC90 442.71 µg/mL), and An. stephensi (LC50 207.06; LC90 408.46 µg/mL), respectively, while Priyangu-synthesized Ag nanoparticles were highly toxic to Cx. quinquefasciatus (LC50 24.91; LC90 45.96 µg/mL), Ae. aegypti (LC50 22.80; LC90 43.23 µg/mL), and An. stephensi (LC50 20.66; LC90 39.94 µg/mL), respectively. Priyangu extract and Ag nanoparticles were found safer to non-target larvivorous fishes, backswimmers, and waterbugs, with LC50 ranging from 1247 to 37,254.45 µg/mL, if compared to target pests. Overall, the current research represents a modern approach integrating traditional botanical pesticides and nanotechnology to the control of larval populations of mosquito vectors, with negligible toxicity against non-target including larvivorous fishes, backswimmers, and waterbugs.