A simplified method for blood feeding, oral infection, and saliva collection of the dengue vector mosquitoes

Sri-in, Chalida; Weng, Shih-Che; Shiao, Shin-Hong; Tu, Wu‐Chun

doi:10.1371/journal.pone.0233618

Cited by 24 publications

(33 citation statements)

References 30 publications

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“…After the blood feeding, each mosquito was examined on a stereo microscope to determine whether it had taken a full meal. Mosquitoes kept at 28 °C and 70% relative humidity under a 12 h:12 h light–dark cycle as previously described [ 31 , 32 ].…”

Section: Methodsmentioning

confidence: 99%

Blood glucose promotes dengue virus infection in the mosquito Aedes aegypti

2021

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Background Dengue fever is the most rapidly spreading mosquito-borne viral disease globally. More than 2.5 billion people live in dengue-endemic areas. Previous studies suggested an interrelationship between diabetes mellitus (DM) and dengue hemorrhagic fever (DHF). Conversely, glycolysis is a critical metabolic pathway for optimal dengue virus (DENV) replication. However, little is known concerning the effect of glucose on DENV replication in mosquitoes. In this study, we investigated the impact of glucose on DENV replication in mosquitoes Aedes aegypti. Methods Mosquitoes (Ae. aegypti UGAL/Rockefeller strain) were orally infected with DENV (serotype 2, 16681 strain) through infectious blood feeding. The DENV infection and transmission rates were determined by examining mosquito bodies and saliva, respectively, for DENV positivity at different time points after infection. In addition, a reverse genetic approach was applied by introducing double-stranded RNA against genes of interest into the mosquitoes to inhibit gene expression. Results Our data revealed a significant increase of DENV genome levels in mosquitoes consuming an infectious blood meal supplemented with glucose, suggesting that blood glucose is an important factor for viral replication. Interestingly, a significant increase of DENV E protein levels was detected in the saliva 4 days faster in mosquitoes that consumed infectious blood meals supplemented with glucose than in those consuming infectious blood meals alone. Furthermore, we perform RNAi to silence AKT or TOR and investigate the molecular mechanism regulating the glucose-mediated enhancement of viral replication. Silencing of AKT or TOR significantly reduced DENV titers in mosquitoes. Conclusions This study suggested that blood glucose is beneficial to DENV replication and that it facilitates virus transmission in mosquitoes via AKT and TOR signaling. Therefore, our results strengthen our understanding of dengue fever and DM co-morbidity and possibly reveal new targets for specific antiviral therapies. Graphical abstract

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Section: Methodsmentioning

confidence: 99%

Blood glucose promotes dengue virus infection in the mosquito Aedes aegypti

2021

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“…Artificial feeding techniques for mosquitoes have been developed over the years (Costa et al, 2013;Sri-in et al, 2020). For example, Luo Yi-Pey ( 2014) calculated for Ae.…”

Section: Discussionmentioning

confidence: 99%

Simplified artificial blood feeding and oral infection method for mosquitoes

Pereira

Carvalho

Silva

et al. 2020

Preprint

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Mosquitoes such as Aedes aegypti, Aedes albopictus and Culex quinquefasciatus are vectors of many pathogens that greatly affect humankind. The maintenance of these mosquitoes in laboratory permit different studies that can help understanding their biology, as well as the vector-pathogen interaction. In addition to sugar meals, the blood feeding is essential for maintenance of the reproductive cycle in several vectors. The main blood sources for many mosquito colonies are direct feeding on live animal or the use of human/animal blood through artificial feeders. However, this latter process has some disadvantages, as artificial feeders can be very laborious for assembly and decontamination. Based on these observations, a simplified technique for feeding and artificial infection was developed with cotton-pads soaked (CS) and blood or blood and viral supernatant to simulate an artificial infection. The efficiency of the CS technique was investigated through the number of mosquitoes fed/infected, when compared to their respective control group. The CS technique, with blood at room temperature, promoted a feeding rate of 61.4% for Ae. albopictus, 70.8% for Cx. quinquefasciatus and 17% for Ae. aegypti. The control group (Hemotec-feeding) presented 47.9%, 16.5% and 59.1% of feeding success, respectively. To improve the CS technique for Ae. aegypti mosquitoes, the procedure was then performed with blood at 38°C, which was possible to observe a feeding rate of 47.3%, in comparison to 53.2% for the control group (Hemotec). When using the CS technique for artificial infection with Mayaro virus, more than 80% of infection was observed for Ae. aegypti and 100% for Ae. albopictus. In the traditional infection technique (glass feeder), the infection rate was 90% (Ae. aegypti) and 96.6% (Ae. albopictus). For Cx. quinquefasciatus, the infection was positive only with the CS technique, resulting in 1 (5%) mosquito infected with Mayaro virus. Our results suggest that this simplified technique of low-cost feeding and easy assembly, offers good results for feeding (maintenance of colonies) and artificial infection of different species of mosquitoes.

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“…aegypti and Ae. albopictus ( Tseng 2003 , Costa-da-Silva et al 2013 , Carvahlo et al 2014 , Gonzales et al 2015 , Gunathilaka et al 2017 , Sri-In et al 2020 ). Another alternative to animal-based membranes is the use of polytetrafluoroethylene (PTFE) tape (‘plumber’s tape’), which resulted in 100% feeding success in Ae.…”

Section: Membranesmentioning

confidence: 99%

“…This method, however, is not an option for many facilities that are located great distances from animal agriculture. A common source used by many facilities is a variety of animal blood that can be purchased from various scientific supply companies ( Bailey et al 1978 , Novak et al 1991 , Tseng 2003 , Lyski et al 2011 , Richards et al 2012 , Sri-In et al 2020 ). The disadvantage of this option is the high price, which can be cost-prohibitive for laboratories with limited budgets.…”

Section: Bloodmentioning

confidence: 99%

“…The elements needed to successfully elicit blood feeding by mosquitoes using artificial methods include the following: blood source, heat source, membrane, and a vessel or substrate to contain the blood. Optional additions to increase blood-feeding success include: host cues (e.g., odors from human skin; Costa-da-Silva et al 2013 , Carvahlo et al 2014 , Gonzales et al 2015 ); a source of carbon dioxide ( McMeniman et al 2014 ); and phagostimulants such as ATP ( Rutledge et al 1964 , Bailey et al 1978 , Gonzales et al 2015 , Sri-In et al 2020 ). Different combinations of each of these components may be more appropriate for certain species and strains of mosquitoes.…”

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confidence: 99%

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The Use of Frozen, Food-Grade Blood to Successfully Maintain Colonies of Four Species of Mosquitoes (Diptera: Culicidae)

Tyler-Julian

Darrisaw

Lloyd

et al. 2021

Journal of Insect Science

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An essential component of all mosquito-rearing activities is the act of blood-feeding the mosquitoes (Diptera: Culicidae). Many options exist for this purpose including live host animals and a diverse array of artificial-feeding methods. Most of the published artificial-feeding methods involve expensive materials, custom-built devices, or are labor-intensive. All of the previously published methods utilize blood sources, which are either expensive, or difficult to obtain. Additionally, much of the research into artificial blood-feeding methods for mosquitoes has focused on two species: Aedes aegypti (Linnaeus) and Aedes albopictus (Skuse). This article presents a modified artificial blood-feeding method that uses affordable and easily sourced materials, does not require any technical knowledge to assemble, and requires minimal time and effort. The combination of inexpensive aluminum plates, Parafilm and polytetrafluoroethylene tape membranes, an electric germination mat, and frozen, food-grade blood produces exceptional feeding rates and abundant egg production. The method has been used for 2 yr at the Lee County Mosquito Control District to successfully maintain laboratory colonies of four species of mosquito: Ae. aegypti , Ae. albopictus , Aedes taeniorhynchus (Wiedemann), and Culex quinquefasciatus (Say). Variations of this method are reported, which can be used for wild and laboratory colonies of multiple species. This modified method is highly accessible for any small-scale mosquito rearing facility with labor or budgetary constraints.

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A simplified method for blood feeding, oral infection, and saliva collection of the dengue vector mosquitoes

Cited by 24 publications

References 30 publications

Blood glucose promotes dengue virus infection in the mosquito Aedes aegypti

Blood glucose promotes dengue virus infection in the mosquito Aedes aegypti

Simplified artificial blood feeding and oral infection method for mosquitoes

The Use of Frozen, Food-Grade Blood to Successfully Maintain Colonies of Four Species of Mosquitoes (Diptera: Culicidae)

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