scholarly journals Transgenic refractory Aedes aegypti lines are resistant to multiple serotypes of dengue virus

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Wei-Liang Liu ◽  
Chia-Wei Hsu ◽  
Shih-Peng Chan ◽  
Pei-Shi Yen ◽  
Matthew P. Su ◽  
...  
Keyword(s):  
Aedes Aegypti ◽  
Dengue Virus ◽  
Blood Meal ◽  
Disease Vectors ◽  
Infection Rates ◽  
Carboxypeptidase A ◽  
Denv Serotypes ◽  
Global Spread ◽  
Lower Infection ◽  
Transgenic Mosquitoes

AbstractThe areas where dengue virus (DENV) is endemic have expanded rapidly, driven in part by the global spread of Aedes species, which act as disease vectors. DENV replicates in the mosquito midgut and is disseminated to the mosquito’s salivary glands for amplification. Thus, blocking virus infection or replication in the tissues of the mosquito may be a viable strategy for reducing the incidence of DENV transmission to humans. Here we used the mariner Mos1 transposase to create an Aedes aegypti line that expresses virus-specific miRNA hairpins capable of blocking DENV replication. These microRNA are driven by the blood-meal-inducible carboxypeptidase A promoter or by the polyubiquitin promoter. The transgenic mosquitoes exhibited significantly lower infection rates and viral titers for most DENV serotypes 7 days after receiving an infectious blood meal. The treatment was also effective at day 14 post infection after a second blood meal had been administered. In viral transmission assay, we found there was significantly reduced transmission in these lines. These transgenic mosquitoes were effective in silencing most of the DENV genome; such an approach may be employed to control a dengue fever epidemic.

scholarly journals A Thioester-Containing Protein Controls Dengue Virus Infection in Aedes aegypti Through Modulating Immune Response

2021 ◽  
Vol 12 ◽  
Author(s):  
Shih-Che Weng ◽  
Hsing-Han Li ◽  
Jian-Chiuan Li ◽  
Wei-Liang Liu ◽  
Chun-Hong Chen ◽  
...  
Keyword(s):  
Immune Response ◽  
Aedes Aegypti ◽  
Dengue Virus ◽  
Blood Meal ◽  
Molecular Mechanisms ◽  
Denv Infection ◽  
Host Factors ◽  
Complement C3 ◽  
Loss Of Function ◽  
Transgenic Mosquitoes

Complement-like proteins in arthropods defend against invading pathogens in the early phases of infection. Thioester-containing proteins (TEPs), which exhibit high similarity to mammalian complement C3, are thought to play a key role in the innate immunity of arthropods. We identified and characterized anti-dengue virus (DENV) host factors, in particular complement-like proteins, in the mosquito Aedes aegypti. Our results indicate that TEP1 limits DENV infection in Ae. aegypti. We showed that TEP1 transcription is highly induced in mosquitoes following DENV infection. Silencing TEP1 resulted in the up-regulation of viral RNA and proteins. In addition, the production of infectious virus particles increased in the absence of TEP1. We generated a transgenic mosquito line with a TEP1 loss-of-function phenotype under a blood meal-inducible promoter. We showed that viral protein and titers increased in transgenic mosquitoes after an infectious blood meal. Interestingly, expression of transcription factor Rel2 and certain anti-microbial peptides (AMPs) were inhibited in transgenic mosquitoes. Overall, our results suggest that TEP1 regulates the immune response and consequently controls the replication of dengue virus in mosquitoes. This finding provides new insight into the molecular mechanisms of mosquito host factors in the regulation of DENV replication.

scholarly journals Use of immunodot blot and multiplex reverse transcriptase-polymerase chain reaction in dengue virus detection in macerates of Aedes aegypti larvae

2012 ◽  
Vol 3 (1) ◽  
pp. 13
Author(s):  
Aline T.A. Chagas ◽  
Michelle D. Oliveira ◽  
Jose M.S. Mezencio ◽  
Eduardo A.M. Silva ◽  
Leandro L. Oliveira ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Aedes Aegypti ◽  
Dengue Virus ◽  
Reverse Transcriptase ◽  
Epidemiological Surveillance ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Denv Serotypes ◽  
Polymerase Chain ◽  
Pcr Techniques

The <em>Dengue virus</em> is the main arbovirus that affects man in terms of morbidity and mortality. The detection of the virus is very important for epidemiological surveillance, so here we propose to standardize and compare the immunodot blot (IDB) and multiplex reverse transcriptase-polymerase chain reaction (M-RT-PCR) techniques to detect and characterize the dengue virus (DENV) serotypes in samples of <em>Aedes aegypti</em> larvae. Thus, the IDB and M-RT-PCR techniques were standardized using macerated samples of larvae collected in nature. The use of monoclonal antibodies in IDB has not shown great results, but DENV detection through this method was possible using polyclonal antibodies. The distinction of serotypes 1, 2 and 3 was carried out by M-RT-PCR.

scholarly journals Differential replication of dengue virus serotypes 2 and 3 in coinfections of C6/36 cells and Aedes aegypti mosquitoes

2014 ◽  
Vol 8 (07) ◽  
pp. 876-884 ◽  
Author(s):  
Diana Carolina Quintero-Gil ◽  
Marta Ospina ◽  
Jorge Emilio Osorio-Benitez ◽  
Marlen Martinez-Gutierrez
Keyword(s):  
Aedes Aegypti ◽  
Dengue Virus ◽  
Cell Viability ◽  
Denv Infection ◽  
Replication Capacity ◽  
Denv Serotypes ◽  
Viral Genomes ◽  
Diphenyltetrazolium Bromide

Introduction: Different dengue virus (DENV) serotypes have been associated with greater epidemic potential. In turn, the increased frequency in cases of severe forms of dengue has been associated with the cocirculation of several serotypes. Because Colombia is a country with an endemic presence of all four DENV serotypes, the aim of this study was to evaluate the in vivo and in vitro replication of the DENV-2 and DENV-3 strains under individual infection and coinfection conditions. Methodology: C6/36HT cells were infected with the two strains individually or simultaneously (coinfection). Replication capacity was evaluated by RT-qPCR, and the effects on cell viability were assessed with an MTT (3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. Additionally, Aedes aegypti mosquitoes were artificially fed the two strains of each serotype individually or simultaneously. The viral genomes were quantified by RT-qPCR and the survival of the infected mosquitoes was compared to that of uninfected controls. Results: In single infections, three strains significantly affected C6/36HT cell viability, but no significant differences were found in the replication capacities of the strains of the same serotype. In the in vivo infections, mosquito survival was not affected, and no significant differences in replication between strains of the same serotype were found. Finally, in coinfections, serotype 2 replicated with a thousandfold greater efficiency than serotype 3 did both in vitro and in vivo. Conclusions: Due to the cocirculation of serotypes in endemic regions, further studies of coinfections in a natural environment would further an understanding of the transmission dynamics that affect DENV infection epidemiology.

scholarly journals The Mosquito Immune System and the Life of Dengue Virus: What We Know and Do Not Know

Pathogens ◽  
2019 ◽  
Vol 8 (2) ◽  
pp. 77 ◽  
Author(s):  
Debica Mukherjee ◽  
Sandeepan Das ◽  
Feroza Begum ◽  
Sweety Mal ◽  
Upasana Ray
Keyword(s):  
Innate Immunity ◽  
Immune System ◽  
Aedes Aegypti ◽  
Dengue Virus ◽  
Aedes Albopictus ◽  
Blood Meal ◽  
Mosquito Vector ◽  
Preventive Strategies ◽  
Immune Control ◽  
The Family

Flaviviruses are largely transmitted to humans by their arthropod vectors such as mosquitoes or ticks. The dengue virus (DENV) is one of the members of the family Flaviviridae and is the causative agent of dengue fever. In the mosquito vector, DENV enters through viremic blood meal and replicates in the mid-gut. Newly formed virion particles circulate to various mosquito organs and get transmitted to the next host in subsequent bites. Aedes aegypti and Aedes albopictus have intricate immune control to allow DENV production at a sub-pathogenic level. In the mosquito, antimicrobial peptides (AMP) and RNA inference (RNAi) are the two main antiviral strategies used against DENV. Apart from innate immunity, mosquito resident microbes play a significant role in modulating DENV replication. In this review, we discuss different immune mechanisms and preventive strategies that act against DENV in two of its vectors: Aedes aegypti and Aedes albopictus.

scholarly journals Sialic Acid Expression in the MosquitoAedes aegyptiand Its Possible Role in Dengue Virus-Vector Interactions

2015 ◽  
Vol 2015 ◽  
pp. 1-16 ◽  
Author(s):  
Jorge Cime-Castillo ◽  
Philippe Delannoy ◽  
Guillermo Mendoza-Hernández ◽  
Verónica Monroy-Martínez ◽  
Anne Harduin-Lepers ◽  
...  
Keyword(s):  
Sialic Acid ◽  
Aedes Aegypti ◽  
Dengue Virus ◽  
Mammalian Cells ◽  
Vector Competence ◽  
Denv Infection ◽  
Viral Disease ◽  
Salivary Gland Extract ◽  
Denv Serotypes ◽  
Mosquito Saliva

Dengue fever (DF) is the most prevalent arthropod-borne viral disease which affects humans. DF is caused by the four dengue virus (DENV) serotypes, which are transmitted to the host by the mosquitoAedes aegyptithat has key roles in DENV infection, replication, and viral transmission (vector competence). Mosquito saliva also plays an important role during DENV transmission. In this study, we detected the presence of sialic acid (Sia) inAedes aegyptitissues, which may have an important role during DENV-vector competence. We also identified genome sequences encoding enzymes involved in Sia pathways. The cDNA forAedes aegyptiCMP-Sia synthase (CSAS) was amplified, cloned, and functionally evaluated via the complementation of LEC29.Lec32 CSAS-deficient CHO cells.AedesCSAS-transfected LEC29.Lec32 cells were able to express Sia moieties on the cell surface. Sequences related toα-2,6-sialyltransferase were detected in theAedes aegyptigenome. Likewise, we identified Sia-α-2,6-DENV interactions in different mosquito tissues. In addition, we evaluated the possible role of sialylated molecules in a salivary gland extract during DENV internalization in mammalian cells. The knowledge of early DENV-host interactions could facilitate a better understanding of viral tropism and pathogenesis to allow the development of new strategies for controlling DENV transmission.

scholarly journals Arbovirus emergence in the temperate city of Córdoba, Argentina, 2009–2018

2019 ◽  
Vol 6 (1) ◽  
Author(s):  
Michael A. Robert ◽  
Daniela T. Tinunin ◽  
Elisabet M. Benitez ◽  
Francisco F. Ludueña-Almeida ◽  
Moory Romero ◽  
...  
Keyword(s):  
Aedes Aegypti ◽  
Dengue Virus ◽  
Disease Transmission ◽  
Data Set ◽  
Denv Serotypes ◽  
Global Problem ◽  
Imported Cases ◽  
The Tropics ◽  
The City ◽  
Autochthonous Transmission

AbstractThe distribution of arbovirus disease transmission is expanding from the tropics and subtropics into temperate regions worldwide. The temperate city of Córdoba, Argentina has been experiencing the emergence of dengue virus, transmitted by the mosquito Aedes aegypti, since 2009, when autochthonous transmission of the virus was first recorded in the city. The aim of this work is to characterize the emergence of dengue and related arboviruses (Zika and chikungunya) in Córdoba since 2009. Herein, we present a data set with all known information about transmission of dengue, Zika, and chikungunya viruses in Córdoba, Argentina from 2009–2018, including what information is known of dengue virus (DENV) serotypes in circulation and origins of imported cases. The data presented in this work will assist researchers in investigating drivers of arbovirus emergence and transmission in Córdoba, Argentina and contribute to a better understanding of the global problem of the expanding distribution of arbovirus disease transmission.

scholarly journals Determining vector competence of Aedes aegypti from Ghana in transmitting dengue virus serotypes 1 and 2

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Michael Amoa-Bosompem ◽  
Daisuke Kobayashi ◽  
Kentaro Itokawa ◽  
Katsunori Murota ◽  
Astri Nur Faizah ◽  
...  
Keyword(s):  
Aedes Aegypti ◽  
Dengue Virus ◽  
Urban Areas ◽  
Vector Competence ◽  
Denv Infection ◽  
West African ◽  
Control Group ◽  
African Countries ◽  
Denv Serotypes ◽  
Serotype 1

Abstract Background Dengue virus (DENV) is a mosquito-borne arbovirus transmitted by Aedes mosquitoes, but is not endemic in all areas where this vector is found. For example, the relatively sparse distribution of cases in West Africa is generally attributed to the refractory nature of West African Aedes aegypti (Ae. aegypti) to DENV infection, and particularly the forest-dwelling Ae. aegypti formosus. However, recent studies have shown these mosquitoes to be competent vectors within some West African countries that have suffered outbreaks in the past, such as Senegal. There is however little information on the vector competence of the Ae. aegypti in West African countries such as Ghana with no reported outbreaks. Methods This study examined the vector competence of 4 Ae. aegypti colonies from urban, semi-urban, and two rural locations in Ghana in transmitting DENV serotypes 1 and 2, using a single colony from Vietnam as control. Midgut infection and virus dissemination were determined by quantitative reverse transcription polymerase chain reaction (qRT-PCR), while the presence and concentration of DENV in the saliva of infectious mosquitoes was determined by the focus forming assay. Results There were significant differences in the colonies’ susceptibility to virus infection, dissemination, and transmission. All examined Ghanaian mosquitoes were refractory to infection by DENV serotype 2, while some colonies exhibited potential to transmit DENV serotype 1. None of the tested colonies were as competent as the control group colony. Conclusions These findings give insight into the possible risk of outbreaks, particularly in the urban areas in the south of Ghana, and highlight the need for continuous surveillance to determine the transmission status and outbreak risk. This study also highlights the need to prevent importation of different DENV strains and potential invasion of new highly vector-competent Ae. aegypti strains, particularly around the ports of entry. Graphic Abstract

scholarly journals Surveillance of dengue virus in individual Aedes aegypti mosquitoes collected concurrently with suspected human cases in Tarlac City, Philippines

2020 ◽  
Vol 13 (1) ◽  
Author(s):  
Jean Claude Balingit ◽  
Thaddeus M. Carvajal ◽  
Mariko Saito-Obata ◽  
Maribet Gamboa ◽  
Amalea Dulcene Nicolasora ◽  
...  
Keyword(s):  
Aedes Aegypti ◽  
Dengue Virus ◽  
Vector Control ◽  
Real Time ◽  
Phylogenetic Analyses ◽  
Simultaneous Detection ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Denv Serotypes ◽  
One Step

Abstract Background Vector control measures are critical for the prevention and reduction of dengue virus (DENV) transmission. Effective vector control is reliant not only on knowledge of mosquito abundance, but also on the timely and accurate detection of mosquito-borne infection. Mosquito-based virus surveillance programs typically rely on pool-based mosquito testing, although whether individual-based mosquito testing is a feasible alternative to this has not been widely studied. Applying an individual-based mosquito testing approach, we conducted a 1-month surveillance study of DENV in adult Aedes aegypti mosquitoes in homes of suspected dengue patients during the 2015 peak dengue season in Tarlac City, Philippines to more accurately assess the mosquito infection rate and identify the DENV serotypes and genotypes concurrently co-circulating in mosquitoes and patients there. Methods We performed a one-step multiplex real-time reverse transcription-polymerase chain reaction (RT-PCR) assay for the simultaneous detection and serotyping of DENV in patients and individual female Ae. aegypti mosquitoes. Additionally, we performed sequencing and phylogenetic analyses to further characterize the detected DENV serotypes in mosquitoes and patients at the genotype level. Results We collected a total of 583 adult Ae. aegypti mosquitoes, of which we individually tested 359 female mosquitoes for the presence of DENV. Ten (2.8%) of the 359 female mosquitoes were positive for the presence of DENV. We detected DENV-1, DENV-2, and DENV-4 in the field-collected mosquitoes, which was consistent with the serotypes concurrently found in infected patients. Sequencing and phylogenetic analyses of the detected DENV serotypes based on the partial sequence of the evelope (E) gene revealed three genotypes concurrently present in the sampled mosquitoes and patients during the study period, namely DENV-1 genotype IV, DENV-2 Cosmopolitan genotype, and DENV-4 genotype II. Conclusions We demonstrated the utility of a one-step multiplex real-time RT-PCR assay for the individual-based DENV surveillance of mosquitoes. Our findings reinforce the importance of detecting and monitoring virus activity in local mosquito populations, which are critical for dengue prevention and control.

Detection of dengue virus in Aedes mosquitoes in Delhi, India

ENTOMON ◽  
2019 ◽  
Vol 44 (3) ◽  
pp. 213-218
Author(s):  
Suresh Chand Kaushik ◽  
Sukhvir Singh ◽  
Purnima Srivastava ◽  
R. Rajendran
Keyword(s):  
Early Intervention ◽  
Early Detection ◽  
Aedes Aegypti ◽  
Dengue Virus ◽  
Infection Rate ◽  
Transmission Season ◽  
Aedes Mosquitoes ◽  
Alternative Approach ◽  
Human Sera ◽  
Minimum Infection Rate

Detection of viruses in human sera particularly in endemic areas is cumbersome and laborious. Therefore, an alternative approach, Immuno-fluorescence assay (IFA) was performed to determine dengue virus (DENV) positivity in mosquitoes. A total of 1055 adult Aedes aegypti female mosquitoes were tested for IFA test against DENV. Minimum infection rate (MIR) for DENV was found higher during August to November 2016 ranging from 10.75 to 20.83. The average yearly MIR was about 6.64. Higher MIR for Ae. aegypti was found in Sarfabad, Noida (12.71) and Khoda Colony, Ghaziabad (11.90). Minimum MIR (4.67) was observed in Sanjay colony (Faridabad). The main contribution of this study resides in the development of a more suitable monitoring system for early detection of viral circulation and to prioritize early intervention in the non-transmission season.

Difusi dan Pola Spasial Sebaran Penyakit Demam Berdarah Dengue (DBD) Di Kota Bandar Lampung

2018 ◽  
Vol 1 (1) ◽  
pp. 87-95
Author(s):  
Nurul Qamila ◽  
Agel Vidian Krama
Keyword(s):  
Public Health ◽  
Spatial Analysis ◽  
Aedes Aegypti ◽  
Dengue Virus ◽  
Spatial Pattern ◽  
Dengue Hemorrhagic Fever ◽  
Health Problem ◽  
Public Health Problem ◽  
Hemorrhagic Fever ◽  
Descriptive Method

Dengue hemorrhagic fever (DHF) is a contagious disease caused by the dengue virus and is transmitted by the mosquito Aedes aegypti (Aa.aegypti). The population is still a public health problem that increases the number of sufferers and also widespread, with population and education. This study aims to reveal the spatial pattern and distribution of Dengue Hemorrhagic Fever (DHF) with the spatial pattern and the spread of Dengue Hemorrhagic Fever (DHF) can result in different locations of these allegations. From the map that can be used for the prevention of Dengue Hemorrhagic Fever (DBD) in Bandar Lampung City. This study aims to reveal the spatial pattern and distribution of Dengue Hemorrhagic Fever (DHF) with the descriptive method and spatial pattern of Dengue Hemorrhagic Fever (DHF) can result in different locations of these allegations. From the map that can be used for the prevention of Dengue Hemorrhagic Fever (DBD) in Bandar Lampung City. Keywords: DHF, Spatial Analysis

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