A fast and inexpensive genotyping system for the simultaneous analysis of human and Aedes albopictus short tandem repeats
Andreu Albó TimorFederica LucatiFrederic BartumeusJenny CanerSanti EscartínSimone MarianiJohn R.B. PalmerMarc Ventura
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Determination of the interactions between hematophagous mosquitoes and their human hosts is of great importance for better understanding the transmission dynamics of mosquito-borne arboviruses and developing effective strategies to mitigate risk. Genetic analysis of human and mosquito DNA can play a key role in this, but commercial kits for human short tandem repeat (STR) genotyping are expensive and do not allow for the simultaneous STR analysis of host and vector DNA. Here, we present an inexpensive and straightforward STR-loci multiplex system capable of simultaneously amplifying Aedes albopictus and human STRs from blood-fed mosquitoes. Additionally, we examine the effect of storage methods and post-feeding time on the integrity of host DNA.Thirty-five STRs (16 human and 19 Ae. albopictus STRs) subdivided in three multiplexes were tested for amplification and scoring reliability. Under laboratory conditions we compared the efficacy of two preservation methods (absolute ethanol vs lysis buffer) on the integrity of host DNA in Ae. albopictus blood meals. We also evaluated the effect of post-feeding time by sacrificing blood-fed mosquitoes at different time intervals after feeding, and we assessed our ability to detect multiple feedings. To determine if the system can be employed successfully under field conditions, we carried out a preliminary study using field-collected Ae. albopictus.All 35 STRs amplified consistently in the laboratory. Lysis buffer performed better than absolute ethanol in terms of allele peak height and clarity of electropherograms. Complete human DNA profiles could be obtained up to 48 h following the blood meal. Analysis of multiple feedings confirmed that peak heights can be used as a proxy to determine post-feeding time and thus derive the number of different people bitten by a mosquito. In the field trial, amplification was successful for 32 STRs. We found human DNA signal in 38 of the 61 field-collected mosquitoes (62%), of which 34 (89%) had ingested a single blood meal, while four (11%) contained double meals.Our new genotyping system allows fast and reliable screening of both host and vector species, and can be further adapted to other mosquito species living in close contact with humans.Keywords:
Aedes albopictus
Blood meal
STR analysis
Multiplex
The behavioral responses of Aedes aegypti (L.) and Aedes albopictus (Skuse) adults to several attractive cues, as reactions to mosquito traps, are compared in the laboratory, and differences in the primary attractive factors for both species are discussed. Target-attacking frequency of unfed Ae. aegypti females was >30 times that of unfed Ae. albopictus females under simulated conditions. Changes in the percentage of trapped mosquitoes under several attractive conditions using commercial mosquito traps showed that Ae. aegypti mosquitoes were trapped 2-3 times faster than Ae. albopictus. For Ae. aegypti, the combination of a visual cue + CO, alone enhanced attractiveness, whereas both a visual cue + CO, as well as a visual cue + octenol enhanced Ae. albopictus. The combination of at least three factors, such as a visual cue, CO2, and a chemical cue is thought to be valuable for trapping and estimating the relative adult population sizes of Ae. aegypti and Ae. albopictus
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Reciprocal and homologous mating experiments between Malaysian Aedes aegypti and Aedes albopictus mosquitoes were conducted in the laboratory. Two methods were employed, namely an artificial mating technique and a natural cage mating technique. The study demonstrated there exists a strong reproductive isolation between Ae. aegypti and Ae. albopictus. Insemination occurred in cross-mating experiments between Ae. aegypti females and Ae. albopictus males and also between Ae. albopictus females and Ae. aegypti males. Cross mating between Ae. aegypti females and Ae. albopictus males produced more eggs than that between Ae. albopictus females and Ae. aegypti males with both artificial mating and natural cage mating techniques. The matings did not result in the production of viable eggs by the females. No embryonation of these eggs was observed when the eggs were bleached. With homologous mating Aedes aegypti produced significantly greater numbers of eggs compared to Aedes albopictus mosquitoes, and all the eggs hatched successfully.
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The behavioral responses of Aedes aegypti (L.) and Aedes albopictus (Skuse) adults to several attractive cues, as reactions to mosquito traps, are compared in the laboratory, and differences in the primary attractive factors for both species are discussed. Target-attacking frequency of unfed Ae. aegypti females was >30 times that of unfed Ae. albopictus females under simulated conditions. Changes in the percentage of trapped mosquitoes under several attractive conditions using commercial mosquito traps showed that Ae. aegypti mosquitoes were trapped 2–3 times faster than Ae. albopictus. For Ae. aegypti, the combination of a visual cue + CO2 alone enhanced attractiveness, whereas both a visual cue + CO2 as well as a visual cue + octenol enhanced Ae. albopictus. The combination of at least three factors, such as a visual cue, CO2, and a chemical cue is thought to be valuable for trapping and estimating the relative adult population sizes of Ae. aegypti and Ae. albopictus in the field.
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Journal Article Physiological Aspects Of Digestion Of The Blood Meal By Aedes Aegypti (Linnaeus) And Culex Fatigans Wiedemann, Get access R. H. Gooding R. H. Gooding 3 3Present address: c/o Department of Microbiology, School of Medicine, Vanderbilt University, Nashville, Tennessee, 37203. Search for other works by this author on: Oxford Academic PubMed Google Scholar Journal of Medical Entomology, Volume 3, Issue 1, 1 April 1966, Pages 53–60, https://doi.org/10.1093/jmedent/3.1.53 Published: 01 April 1966
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Adult female Aedes albopictus and Aedes aegypti mosquitoes were exposed to Orungo, an orbivirus, by intrathoracic inoculation and oral ingestion. After 6 days, virus was demonstrated in 1 orally exposed mosquito, but only inoculated mosquitoes transmitted the virus to blood droplets after an extrinsic incubation of 6–10 days. Transmission attempts utilizing infant mice were negative.
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Aedes aegypti plays an important role in the transmission of several arboviruses of medical importance. The availability of information on the blood-feeding preferences of mosquito vectors is a critical step in the understanding of the transmission of human pathogens and implementation of control strategies. In Senegal, no data currently exist on the feeding pattern of Ae. aegypti in urban areas. To fill this gap, Ae. aegypti blood-fed females were collected in five localities by aspiration and using BG Sentinel 2 traps. Collections were carried out monthly between July and November 2019 inside and outside human dwellings. The origin of the blood meal of Ae. aegypti females were identified by an ELISA technique. A total of 1,710 blood-engorged females were examined and showed that Ae. aegypti preferentially fed on human with 78.6% of the identified blood meals. The other blood meals were from animals including dog, cat, horse, cattle, sheep, and rat. This is the first report on the feeding behavior of Ae. aegypti in urban settings in West Africa. It demonstrated that this species is highly anthropophilic.
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The polygamous behavior of male Aedes aegypti (L.) and Ae. albopictus (Skuse) was investigated by co-habiting a newly-emerged male and females in a 30 cm3 cage (1 male: 20 females) for up to 5 consecutive days. As determined by insemination rates, the results indicated that one Ae. aegypti and Ae. albopictus male could successfully mate with 1.10 (0-4), 4.10 (1-8), 5.40 (4-8), 5.10 (2-8), 5.15 (3-9) and 0.20 (0-3), 1.70 (0-3), 2.35 (1-4), 2.30 (0-4), 2.35 (1-4) Ae. aegypti and Ae. albopictus females, respectively on day 1,2,3,4 and 5 consecutively. The possibly significant role of their polygamy in relation to dengue virus transmission is discussed.
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Plasmodium gallinaceum
Blood meal
Avian Malaria
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