We used a histologic technique to study multiple blood feeding in a single gonotrophic cycle by engorged Aedes aegypti (L.) that were collected weekly for 2 yr from houses in a rural village in Thailand (n = 1,891) and a residential section of San Juan, Puerto Rico (n = 1,675). Overall, mosquitoes from Thailand contained significantly more multiple meals (n = 1,300, 42% double meals, 5% triple meals) than mosquitoes collected in Puerto Rico (n = 1,156, 32% double meals, 2% triple meals). The portion of specimens for which frequency of feeding could not be determined was 31% at both sites. We estimated that on average Ae. aegypti take 0.76 and 0.63 human blood meals per day in Thailand and Puerto Rico, respectively. However, frequency of multiple feeding varied among houses and, in Puerto Rico, the neighborhoods from which mosquitoes were collected. In Thailand 65% of the mosquitoes fed twice on the same day, whereas in Puerto Rico 57% took multiple meals separated by > or = 1 d. At both sites, the majority of engorged specimens were collected inside houses (Thailand 86%, Puerto Rico 95%). The number of blood meals detected was independent of where mosquitoes were collected (inside versus outside of the house) at both sites and the time of day collections were made in Puerto Rico. Feeding rates were slightly higher for mosquitoes collected in the afternoon in Thailand. Temperatures were significantly higher and mosquitoes significantly smaller in Thailand than in Puerto Rico. At both sites female size was negatively associated with temperature. Rates of multiple feeding were associated positively with temperature and negatively with mosquito size in Thailand, but not in Puerto Rico. Multiple feeding during a single gonotrophic cycle is a regular part of Ae. aegypti biology, can vary geographically and under different climate conditions, and may be associated with variation in patterns of dengue virus transmission.

https://bioone.org/journals/journal-of-medical-entomology/volume-37/issue-1/0022-2585-37.1.89/Longitudinal-Studies-of-Aedes-aegypti-Diptera--Culicidae-in-Thailand/10.1603/0022-2585-37.1.89.pdf

Longitudinal Studies of Aedes aegypti (Diptera: Culicidae) in Thailand and Puerto Rico: Blood Feeding Frequency

Background
The survival of adult female Aedes mosquitoes is a critical component of their ability to transmit pathogens such as dengue viruses. One of the principal determinants of Aedes survival is temperature, which has been associated with seasonal changes in Aedes populations and limits their geographical distribution. The effects of temperature and other sources of mortality have been studied in the field, often via mark-release-recapture experiments, and under controlled conditions in the laboratory. Survival results differ and reconciling predictions between the two settings has been hindered by variable measurements from different experimental protocols, lack of precision in measuring survival of free-ranging mosquitoes, and uncertainty about the role of age-dependent mortality in the field.

/pdf/modelling-adult-aedes-aegypti-and-aedes-albopictus-survival-2qjjpumkcj.pdf

Modelling adult Aedes aegypti and Aedes albopictus survival at different temperatures in laboratory and field settings

Adult female Aedes aegypti (L.), the vector of dengue and yellow fever viruses, have an affinity for feeding on human blood and a tendency to forego feeding on sugar. This observation challenges two tenets of mosquito biology: (1) mosquitoes imbibe plant carbohydrates for synthesis of energy reserves and blood for reproduction and (2) egg production is reduced when mosquitoes feed on human blood compared with blood from other species. Sub-optimal amounts of the amino acid isoleucine in human blood (particularly free isoleucine in plasma) are thought to be responsible for lowered egg production when human blood is ingested. We tested the hypothesis that feeding on human blood is associated with a selective advantage for Ae. aegypti and is an underlying reason for this mosquito's intimate and epidemiologically important relationship with human beings. Our five experiments examined the effects of different isoleucine concentrations on accumulated energy reserves, frequency of host contact, survival, and egg production. When mosquitoes imbibed blood meals over a 7- to 10-d period and were not fed sugar, increased isoleucine concentration decreased energy reserves and did not increase egg production. Aedes aegypti took smaller but more frequent blood meals when feeding on a low-isoleucine human host daily compared with a high-isoleucine mouse host. Previous reports that isoleucine enhances egg production were confirmed only when females were fed sugar, an unusual behavior for most domestic Ae. aegypti populations. Females fed human blood and water had greater age-specific survival (l(x)), reproductive output (m(x)), and cumulative net replacement (R0) than cohorts fed human blood plus sugar or isoleucine-rich mouse blood with or without access to sugar. The unique isoleucine concentration of human blood is associated with Ae. aegypti's unusual propensity to feed preferentially and frequently on humans--a behavior that increases this mosquito's fitness, synthesis of energy reserves, and contact with human hosts, making it an especially effective disseminator of human pathogens.

Why Do Female Aedes aegypti (Diptera: Culicidae) Feed Preferentially and Frequently on Human Blood?

Dengue is a disease that has undergone significant expansion over the past hundred years. Understanding what factors limit the distribution of transmission can be used to predict current and future limits to further dengue expansion. While not the only factor, temperature plays an important role in defining these limits. Previous attempts to analyse the effect of temperature on the geographic distribution of dengue have not considered its dynamic intra-annual and diurnal change and its cumulative effects on mosquito and virus populations. Here we expand an existing modelling framework with new temperature-based relationships to model an index proportional to the basic reproductive number of the dengue virus. This model framework is combined with high spatial and temporal resolution global temperature data to model the effects of temperature on Aedes aegypti and Ae. albopictus persistence and competence for dengue virus transmission. Our model predicted areas where temperature is not expected to permit transmission and/or Aedes persistence throughout the year. By reanalysing existing experimental data our analysis indicates that Ae. albopictus, often considered a minor vector of dengue, has comparable rates of virus dissemination to its primary vector, Ae. aegypti, and when the longer lifespan of Ae. albopictus is considered its competence for dengue virus transmission far exceeds that of Ae. aegypti. These results can be used to analyse the effects of temperature and other contributing factors on the expansion of dengue or its Aedes vectors. Our finding that Ae. albopictus has a greater capacity for dengue transmission than Ae. aegypti is contrary to current explanations for the comparative rarity of dengue transmission in established Ae. albopictus populations. This suggests that the limited capacity of Ae. albopictus to transmit DENV is more dependent on its ecology than vector competence. The recommendations, which we explicitly outlined here, point to clear targets for entomological investigation.

/pdf/global-temperature-constraints-on-aedes-aegypti-and-ae-2xkd0xiv1v.pdf

Global temperature constraints on Aedes aegypti and Ae. albopictus persistence and competence for dengue virus transmission

Feeding strategies of anthropophilic mosquitoes result in increased risk of pathogen transmission

Life table studies were performed in 1996 with Aedes aegypti (L.) during the low (cool/dry) and high (hot/rainy) dengue virus transmission seasons in Puerto Rico. Mated adult females from field-collected pupae were placed individually in cages and divided into 2 treatment groups: one was fed only human blood and the other human blood plus a 10% sucrose solution. Survival and number of eggs laid were recorded daily for each female. During both seasons, age specific survivorship was higher for the blood plus sugar group, groups fed only human blood had higher reproductive outputs (mx), and net replacement rates (Ro) for blood only groups were higher than for those fed blood plus sugar. Intrinsic rates of growth (r) were the same for both treatments during the low (cool/dry) transmission season, but higher for the blood-only treatment during the high (hot/rainy) transmission season. Our results indicate that feeding on only human blood provides an evolutionary advantage to Ae. aegypti females in Puerto Rico. These results are similar to those from an earlier study carried out with Ae. aegypti in Thailand; the advantage of feeding on human blood does not seem to be restricted to a particular geographic region. We also found that the benefits associated with human feeding persist through epidemiologically different times of the year. We conclude that feeding on human blood is reproductively beneficial for Ae. aegypti, which may increase their contact with human hosts, and therefore may influence their vectorial capacity for dengue viruses through frequent feeding on blood.

Life table study of Aedes aegypti (Diptera: Culicidae) in Puerto Rico fed only human blood versus blood plus sugar.

Male and female Aedes aegypti fed a 10% sucrose solution and/or blood were tested to determine the duration of fructose detection in their bodies and the volume of sugar solution they ingested. The limit of detection of fructose by the cold anthrone test was investigated in a series of experiments. Results were applied to the interpretation of sugar feeding by Ae. aegypti collected inside houses in Puerto Rico during times of low (cool season) and high (hot season) dengue transmission in 1996. We conclude that, under our experimental conditions, the cold anthrone test can detect a 10% sucrose solution in male and female Ae. aegypti up to approximately 4 days after ingestion, even the smallest volumes of 10% sucrose solution ingested by experimental mosquitoes are detectable, the test is sensitive enough to detect 0.6 microgram of fructose, and the cutoff point for defining positive fructose values in field-collected females should be based on blood-engorged specimens. We confirmed that female Ae. aegypti collected from natural resting sites inside houses in Puerto Rico seldom, compared to males, contain detectable amounts of fructose.

An experimental study on the detection of fructose in Aedes aegypti

Laboratory experiments suggest that utilization of blood rather than natural sugar sources for energetic needs affords female Aedes aegypti a reproductive advantage over conspecifics that use sugar. To test this hypothesis under field conditions, we carried out a mark-release-recapture study in Florida, PR. Adult females (F1) reared from field-collected eggs were provided with a diet of human blood alone or human blood plus a 20% honey solution before their release. Backpack aspirators were used to collect mosquitoes from release houses for 5 consecutive days beginning the 2nd day after release. Survival was estimated from the slope of the regression line of the log-transformed daily number of recaptures for each treatment group. To compare fecundity of the treatment groups, each recaptured female was dissected, ovaries were removed, oocytes counted, and Christophers' stages of oocyte development scored. Recapture rates were 30% for the blood-only group and 23% for blood plus honey group. The daily survival rate of the blood-only group (55%) was not statistically different from that of the blood plus honey group (69%) (t = 0.32, P > 0.05). By analysis of variance, fecundity (average number of stage III-V oocytes) was significantly higher in the females fed human blood alone (n = 103, 109 oocytes/female) than in the group fed on blood and honey (n = 50, 95 oocytes/female) (P = 0.0007). The observed gonotrophic cycle length of the recaptured females ranged from 3 to 7 days. Results from our field study are consistent with laboratory life-table experiments that suggest feeding exclusively on human blood provides a reproductive advantage for female A. aegypti.

/pdf/increased-fecundity-of-aedes-aegypti-fed-human-blood-before-577amovu1n.pdf

Increased fecundity of Aedes aegypti fed human blood before release in a mark-recapture study in Puerto Rico.

Survival of adult Aedes aegypti (L.) was studied in Thailand (1995) and Puerto Rico (1996) during periods of high and low dengue virus transmission. Resting males and females were collected inside houses by aspiration. Females were separated into different cages by their degree of engorgement and ovarian development. Teneral adults were obtained from pupae collected from natural breeding sites. All mosquitoes were given access to water, held at ambient temperature in the shade, and their survival monitored daily. We calculated median survival for each stage to estimate when mosquitoes had to feed again or die. No differences in survival between seasons were observed in Thailand. In Puerto Rico, except for wild males, survival was longer in the cool/dry season than in the hot/rainy season, indicating that mosquitoes may need to feed more frequently during the high than low dengue transmission season. During both study periods and at both sites, blood-engorged females survived as long or longer than mosquitoes in other gonotrophic or developmental stages. Except in Puerto Rico during the cool season, when females had a relatively high probability of surviving 3-4 d without feeding, females needed to feed approximately every other day to avoid death caused by starvation. Our results indicate that in some regions, there are seasonal differences in the length of time female Ae. aegypti can survive without feeding, females with a blood meal can survive for a longer time than those without blood, and teneral males can live longer without food than teneral females.

Adriana Costero

Papers

Life table study of Aedes aegypti (Diptera: Culicidae) in Puerto Rico fed only human blood versus blood plus sugar.

An experimental study on the detection of fructose in Aedes aegypti

Increased fecundity of Aedes aegypti fed human blood before release in a mark-recapture study in Puerto Rico.

Survival of starved Aedes aegypti (Diptera: Culicidae) in Puerto Rico and Thailand.