Field evaluation of the response of Aedes albopictus (Stegomyia albopicta) to three oviposition attractants and different ovitrap placements using black and clear autocidal ovitraps in a rural area of Same,Timor‐Leste

Known oviposition attractants or stimulants were compared, singly and in combination, using inexpensive autocidal ovitraps designed to trap emerging adults, in a rural area of Timor‐Leste during the dry season. In this area, the dengue vector Aedes albopictus (Stegomyia albopicta) Skuse (Diptera: Culicidae) was abundant, but Aedes aegypti (Stegomyia aegypti) L. was not detected. The attractants were: (a) a compound found in Aedes eggs (dodecanoic acid); (b) components of nitrogen, phosphorous and potassium‐based (NPK) fertilizer, and (c) infusions of discarded cigarette butts. A solution of ammonium phosphate and potassium nitrate was significantly more attractive to gravid Ae. albopictus than water only. Dodecanoic acid and cigarette butt infusions were not significantly more attractive than the control; however, they attracted various other Diptera and many non‐culicid larvae developed in ovitraps in which these substances were used; thus, the presence of eggs or larvae of other species may have deterred Aedes oviposition. Significantly more Aedes eggs were found in ovitraps under vegetation than in ovitraps placed inside houses or against external walls. Clear‐sided ovitraps in which black mesh was placed over a black ring floating on the water surface collected significantly fewer eggs than black ovitraps with identically placed mesh and rings.     

Electrophysiological and behavioural response of Aedes albopictus to n‐heinecosane,an ovipositional pheromone of Aedes aegypti

Aedes aegypti (L.) and Aedes albopictus (Skuse) (Diptera: Culicidae) are highly anthropophilic mosquito species and potential vectors of dengue and yellow fever. The location of suitable sites for oviposition requires a set of visual, tactile, and olfactory cues that influence females before they lay their eggs. In this study, the effect of n‐heneicosane, a recognized oviposition pheromone of Ae. aegypti, on the olfactory receptors of the antennae of Ae. aegypti and Ae. albopictus was studied using electroantennographic detection coupled to gas chromatography (GC‐EAD). A significant electroantennographic response to n‐heneicosane in adult females of both mosquito species was observed. In addition, gravid Ae. albopictus females laid more eggs in substrate treated with n‐heneicosane at 0.1, 1, or 10 p.p.m. than in the control, denoting oviposition attractancy. Conversely, at 30, 50, 100, and 200 p.p.m., more eggs were laid in the control substrate, indicating oviposition repellency. Analysis of the larval cuticle by GC and mass spectrometry confirmed the presence of n‐heneicosane in the cuticles of Ae. albopictus larvae. The species‐specific role of n‐heneicosane as an oviposition pheromone in Ae. aegypti and its significance as a behaviour modifier of Ae. albopictus in breeding sites is discussed.     

A lethal ovitrap‐based mass trapping scheme for dengue control in Australia: II. Impact on populations of the mosquito Aedes aegypti

In Cairns, Australia, the impacts on Aedes aegypti L. (Diptera: Culicidae) populations of two types of ‘lure & kill’ (L&K) lethal ovitraps (LOs), the standard lethal ovitrap (SLO) and the biodegradable lethal ovitrap (BLO) were measured during three mass‐trapping interventions. To assess the efficacy of the SLO, two interventions (one dry season and one wet season) were conducted in three discrete areas, each lasting 4 weeks, with the following treatments: (i) SLOs (>200 traps, ∼4/premise), BG‐sentinel traps (BGSs; ∼15, 1/premise) and larval control (container reduction and methoprene treatment) and (ii) larval control alone, and (iii) untreated control. Female Ae. aegypti populations were monitored for 4 weeks pre‐ and post‐treatment in all three areas using BGSs and sticky ovitraps (SOs) or non‐lethal regular ovitraps (ROs). In the dry season, 206 SLOs and 15 BGSs set at 54 and 15 houses, respectively, caught and killed an estimated 419 and 73 female Ae. aegypti, respectively. No significant decrease in collection size of female Ae. aegypti could be attributed to the treatments. In the wet season, 243 SLOs and 15 BGSs killed ∼993 and 119 female Ae. aegypti, respectively. The mean number of female Ae. aegypti collected after 4 weeks with SOs and BGSs was significantly less than the control (LSD post‐hoc test). The third mass‐trapping intervention was conducted using the BLO during the wet season in Cairns. For this trial, three treatment areas were each provided with BLOs (>500, ∼4/premise) plus larval control, and an untreated control area was designated. Adult female Ae. aegypti were collected for 4 weeks pre‐ and post‐treatment using 15 BGSs and 20 SOs. During this period, 53.2% of BLOs contained a total of 6654 Ae. aegypti eggs. Over the intervention period, collections of Ae. aegypti in the treatment areas were significantly less than in the control area for BGSs but not SOs. An influx of relatively large numbers of young females may have confounded the measurement of changes in populations of older females in these studies. This is an important issue, with implications for assessing delayed action control measures, such as LOs and parasites/pathogens that aim to change mosquito age structure. Finally, the high public acceptability of SLOs and BLOs, coupled with significant impacts on female Ae. aegypti populations in two of the three interventions reported here, suggest that mass trapping with SLOs and BLOs can be an effective component of a dengue control strategy.     

Influence of shading and pedestrian traffic on the preference of Aedes (Stegomyia) aegypti (Diptera: Culicidae) for oviposition microenvironments

Aedes aegypti mosquitoes are highly adaptable to abiotic stimuli. To evaluate the influence of shading and pedestrian traffic on the preference of Ae. aegypti for oviposition microenvironments, 20 sites were sampled weekly using ovitraps within the perimeter of Universidade Federal do Espírito Santo, located in São Mateus, Espírito Santo, Brazil. A spatial and statistical analysis was performed in order to assess the relationship between shading time, pedestrian traffic, and the presence of biological forms of Ae. aegypti. A temporal analysis of temperature and precipitation influence on oviposition was also made. Between June, 2013 and June, 2014, 7,362 Ae. aegypti eggs were collected. Over a 12‐month period, we made weekly collections of Ae. aegypti eggs from ovitraps. Pedestrian traffic and shading time influenced the number of positive ovitraps; precipitation and temperature were correlated with the number of positive ovitraps (p <0.05). We conclude that the influence of temperature and precipitation was not significant for the oviposition index, and the frequency of oviposition was directly proportional to the number of individuals moving close to the traps during periods of greater shading.     

Multi‐scale analysis of the associations among egg,larval and pupal surveys and the presence and abundance of adult female Aedes aegypti (Stegomyia aegypti) in the city of Merida,Mexico

Despite decades of research, there is still no agreement on which indices of Aedes aegypti (Stegomyia aegypti) (Diptera: Culicidae) presence and abundance better quantify entomological risk for dengue. This study reports the results of a multi‐scale, cross‐sectional entomological survey carried out in 1160 households in the city of Merida, Mexico to establish: (a) the correlation between levels of Ae. aegypti presence and abundance detected with aspirators and ovitraps; (b) which immature and egg indices correlate with the presence and abundance of Ae. aegypti females, and (c) the correlations amongst traditional Aedes indices and their modifications for pupae at the household level and within medium‐sized geographic areas used for vector surveillance. Our analyses show that ovitrap positivity was significantly associated with indoor adult Ae. aegypti presence [odds ratio (OR) = 1.50; P = 0.03], that the presence of pupae is associated with adult presence at the household level (OR = 2.27; P = 0.001), that classic Aedes indices are informative only when they account for pupae, and that window screens provide a significant level of protection against peridomestic Ae. aegypti (OR = 0.59; P = 0.02). Results reinforce the potential of using both positive collections in outdoor ovitraps and the presence of pupae as sensitive indicators of indoor adult female presence.     

Lethal ovitrap deployment for Aedes aegypti control: potential implications for non‐target organisms

In Australia, dengue control combines source reduction with lethal ovitraps to reduce Aedes aegypti populations during outbreaks. Lethal ovitraps are considered a sustainable and environmentally friendly method of controlling container‐inhabiting mosquitoes, however, to‐date, this claim has not been quantified. This study assesses the potential impact of lethal ovitraps on non‐target organisms when used to control Ae. aegypti in tropical Australia. For retention of specimens, we substituted standard sticky ovitraps for lethal ovitraps. We collected 988 Ae. aegypti and 44,132 non‐target specimens over 13 months from 16 sites. Although Ae. aegypti comprised only 2.2% of the total collection, they were were the eighth most dominant taxa collected, on the 93^rd percentile. Of the non‐target organisms, Collembola were the dominant taxa, 44.2%, with 36.8% and 10.5% Diptera and Hymenoptera, respectively. Of the Dipterans, 61% were family Phoridae. Lethal ovitraps were visited by 90 insect or invertebrate families in total. Ovitraps are attractive to Collembola, Phoridae, Sciaridae, Formicidae, and Culicidae, with minimal attraction by Apidae and other commonly monitored non‐target organisms. For container‐inhabiting mosquitoes, LOs are cost effective operationally, requiring minimal staff resources for placement and retrieval.     

Aedes albopictus oviposits with other Aedes species in artificial oviposition cups: a case study in Knox County,Tennessee, U.S.A.

Interspecific associations between two mosquito species can lead to effects such as competition, species displacement, and species stability. To better understand Aedes albopictus (Skuse) and other Aedes species, we monitored eggs in artificial oviposition cups (ovitraps) within Knox County, TN, U.S.A., during the 2016 and 2017 mosquito seasons. In 2016, one black and one white ovitrap were placed at 18 sites for 21 weeks, while in 2017 black and white ovitraps baited with grass‐infused or deionized water were placed at 11 sites for nine weeks. Eggs were identified to species and resulting counts were used to determine the degree of interspecific association using Cole's coefficients (C₇) and the degree of heterogeneity across space and time using generalized linear mixed models (GLMM). Here, Aedes mosquitoes are generally ovipositing in black cups with grass‐infused water, and Ae. albopictus eggs co‐occurred with other Aedes species more often than would be expected. Finding a positive significant interspecific association between Ae. albopictus and other Aedes eggs suggests that methods used to control Ae. albopictus may also control other Aedes mosquitoes. Finding that Ae. albopictus co‐occurs with other Aedes mosquitoes warrants additional research to evaluate outcomes associated with co‐occurrence within the study area.     

Elimination of Aedes aegypti in northern Australia, 2004–2006

The Northern Territory (NT) of Australia is currently free of the dengue mosquito Aedes (Stegomyia) aegypti (L). However, on 17 February 2004, two Ae. aegypti adults were captured in two routine CO₂‐baited encephalitis virus surveillance traps in Tennant Creek, located 990 km south of Darwin in the NT. The detection triggered an immediate survey and control response undertaken by the NT Department of Health and Community Services, followed by a Commonwealth of Australia‐funded Ae. aegypti elimination program. This report details the methods and results of the detection and subsequent elimination activities that were carried out between 2004 and 2006, returning the NT to its dengue vector‐free status. There have been very few successful Ae. aegypti elimination programs in the world. This purposeful mosquito elimination for Australia was officially declared on 5 April 2006.     

Efficacy of sticky and standard ovitraps for Aedes aegypti in Trinidad,West Indies

The double sticky trap (DST) is described for the first time and is evaluated along with standard ovitraps and sticky traps (STs) to determine population densities of Ae. aegypti in the urban township of St. Augustine and the rural community of Tamana, Trinidad, West Indies. Ten houses were selected at each study site. At each of the ten houses, one ovitrap, one ST, and one DST were placed using the criteria established for placement of ovitraps. The results showed the three trapping methods successfully collected Ae. aegypti mosquitoes. All three traps collected significantly more adults or eggs in St. Augustine than in Tamana. DSTs collected 2,286 adults from St. Augustine vs 316 adults from Tamana (p<0.002), STs collected 1,480 and 220 adults, respectively (p<0.01), and the ovitraps collected 2,735 and 517 eggs, respectively from St. Augustine and Tamana (p<0.002). Based on these results, the DSTs collected significantly (P<0.02) more adults than the STs. The DSTs and STs collected both adult and immature stages which can be used for toxicology, virology, and PCR studies and are suitable alternative Ae. aegypti surveillance tools for the Caribbean and Latin American region.     

Field evaluation of baited traps for surveillance of Aedes japonicus japonicus in Switzerland

The efficacy of Centers for Disease Control (CDC) miniature light traps and ovitraps was tested in the outskirts of the city of Zurich in Switzerland for their use in the surveillance of Aedes (Hulecoeteomyia) japonicus japonicus (Theobald) (Diptera: Culicidae), the invasive Asian bush mosquito. Sets of single CDC traps were run overnight (n = 18) in three different environments (forest, suburban and urban) in 3 × 3 Latin square experimental designs. Traps were baited with: (a) carbon dioxide (CO₂); (b) CO₂ plus light, or (c) CO₂ plus lure blend [Combi FRC 3003 (iGu^®)]. At the same locations, mosquito eggs were collected weekly using standard ovitraps baited with different infusions (oak, hay or tap water) and equipped with different oviposition substrates (a block of extruded polystyrene, a germination paper strip or a wooden stick). Data were analysed using Poisson and negative binomial general linear models. The use of light (P < 0.001) or lure (P < 0.001) significantly increased the attractiveness of CDC traps baited with CO₂. Oak and hay infusions did not increase the attractiveness of ovitraps compared with standing tap water (P > 0.05), and extruded polystyrene blocks were preferred as an oviposition substrate over wooden sticks (P < 0.05) and seed germination paper (P < 0.05). Carbon dioxide‐baited CDC miniature light traps complemented with light or iGu^® lure and ovitraps containing standing tap water and polystyrene oviposition blocks can be considered as efficient and simple tools for use in Ae. j. japonicus surveillance programmes.     

Oviposition substrate selection by Florida mosquitoes in response to pathogen‐infected conspecific larvae

The impact of the presence of larval mosquito pathogens with potential for biological control on oviposition choice was evaluated for three mosquito species/pathogen pairs present in Florida. These included Aedes aegypti infected with Edhazardia aedis, Aedes albopictus infected with Vavraia culicis, and Culex quinquefasciatus infected with Culex nigripalpus nucleopolyhedrovirus (CuniNPV). Two‐choice oviposition bioassays were performed on each host and pathogen species with one oviposition cup containing infected larvae and the other cup containing uninfected larvae (control). Both uninfected and E. aedis‐infected female Ae. aegypti laid significantly fewer eggs in oviposition cups containing infected larvae. Uninfected gravid female Ae. albopictus and Cx. quinquefasciatus oviposited equally in cups containing uninfected larvae or containing larvae infected with V. culicis or CuniNPV, respectively. Gravid female Ae. albopictus infected with V. culicis did not display ovarian development and did not lay eggs. The decreased oviposition by gravid Ae. aegypti in containers containing E. aedis‐infected larvae may indicate that the infected larvae produce chemicals deterring oviposition.     

Comparative efficacy of existing surveillance tools for Aedes aegypti in Western Kenya

All traditional surveillance techniques for Aedes aegypti have been developed for the cosmopolitan domestic subspecies Ae. aegypti aegypti, and not the sylvatic subspecies, Ae. aegypti formosus. The predominant form in Western Kenya is Ae. aegypti formosus that is rarely associated with human habitations but is linked to transmission of sylvatic dengue virus strains. We compared five surveillance methods for their effectiveness in sampling Ae. aegypti formosus with the goal of determining a sustainable surveillance strategy in Kenya. The methods included larval and pupal surveys, oviposition trapping, BG‐Sentinel trapping, resting boxes, and backpack aspirations. Larval and pupal surveys collected the highest number of Ae. aegypti formosus (51.3%), followed by oviposition traps (45.7%), BG‐Sentinel traps (3.0%), and zero collected with either backpack aspiration or resting box collections. No Ae. aegypti formosus larvae or pupae were found indoors. The results indicate that oviposition traps and outdoor larval and pupal surveys were better surveillance methods for Ae. aegypti formosus in Western Kenya.     

Susceptibility of Florida Aedes aegypti and Aedes albopictus to dengue viruses from Puerto Rico

Locally acquired dengue cases in the continental U.S. are rare. However, outbreaks of dengue‐1 during 2009, 2010, and 2013 in Florida and dengue‐1 and −2 in Texas suggest vulnerability to transmission. Travel and commerce between Puerto Rico and the U.S. mainland is common, which may pose a risk for traveler‐imported dengue cases. Mosquitoes were collected in Florida and used to evaluate their susceptibility to dengue viruses (DENV) from Puerto Rico. Aedes aegypti and Ae. albopictus were susceptible to virus infection with DENV‐1 and −2. No significant differences were observed in rates of midgut infection or dissemination between Ae. aegypti or Ae. albopictus for DENV‐1 (6–14%). Aedes aegypti was significantly more susceptible to midgut infection with DENV‐2 than Ae. albopictus (Ae. aegypti, ∼28%; Ae. albopictus, ∼9%). The dissemination rate with dengue‐2 virus for Ae. aegypti (23%) was greater than Ae. albopictus (0%), suggesting that Ae. albopictus is not likely to be an important transmitter of the DENV‐2 isolate from Puerto Rico. These results are discussed in light of Florida's vulnerability to DENV transmission.     

Determining the spatial autocorrelation of dengue vector populations: influences of mosquito sampling method,covariables, and vector control

We investigated spatial autocorrelation of female Aedes aegypti L. mosquito abundance from BG‐Sentinel trap and sticky ovitrap collections in Cairns, north Queensland, Australia. BG‐Sentinel trap collections in 2010 show a significant spatial autocorrelation across the study site and over a smaller spatial extent, while sticky ovitrap collections only indicate a non‐significant, weak spatial autocorrelation. The BG‐Sentinel trap collections were suitable for spatial interpolation using ordinary kriging and cokriging techniques. The uses of Premise Condition Index and potential breeding container data have helped improve our prediction of vector abundance. Semiovariograms and prediction maps indicate that the spatial autocorrelation of mosquito abundance determined by BG‐Sentinel traps extends farther compared to sticky ovitrap collections. Based on our data, fewer BG‐Sentinel traps are required to represent vector abundance at a series of houses compared to sticky ovitraps. A lack of spatial structure was observed following vector control treatment in the area. This finding has implications for the design and costs of dengue vector surveillance programs.     

Evaluation of collection methods for Culex quinquefasciatus,Aedes aegypti,and Aedes simpsoni in northeastern Tanzania

In East Africa, significant morbidity and mortality are caused by infections spread by Culex quinquefasciatus and Aedes aegypti. Sticky traps have been shown to be effective tools for sampling populations of Aedes mosquitoes and have been found to catch Cx. quinquefasciatus. Thus, they could potentially be used to sample populations of this species. This study compared Sticky ovitraps (SO) and MosquiTraps (MQT) with the CDC Gravid trap (CDC‐GT) for collection of Culex and Aedes mosquito populations in Tanzania. A follow‐up experiment was carried out using traps set for a 24‐h period to accommodate the oviposition habits of Aedes aegypti and Ae. simpsoni s.l. mosquitoes. The results showed that the CDC‐GT caught significantly more Cx. quinquefasciatus and Ae. aegypti than the SO or MQT, but there was no significant difference in the number of mosquitoes caught between the two sticky traps or of Ae. simpsoni s.l. caught among the three trap types. The results suggest that CDC‐GTs are the most appropriate in sampling of Cx. quinquefasciatus. Although CDC‐GTs collected more Ae. aegypti than the sticky traps, the simplicity and cost benefit of sticky traps facilitates large scale studies. All three trap types should be considered for monitoring Aedes mosquitoes.     

Seasonal population dynamics and the genetic structure of the mosquito vector Aedes aegypti in São Paulo,Brazil

Population genetic studies of insect vectors can generate knowledge to improve epidemiological studies focused on the decrease of pathogen transmission. In this study, we used nine SNPs across the Aedes aegypti genome to characterize seasonal population variations of this important dengue vector. Mosquito samples were obtained by ovitraps placed over Botucatu SP from 2005 to 2010. Our data show that, regardless of the large variation in mosquito abundance (deduced from the number of eggs obtained from ovitraps), the effective population size remained stable over the years. These results suggest that Ae. aegypti is able to maintain a sufficiently large active breeding population during the dry season to keep genetic frequencies stable. These results open new perspectives on mosquito survey and control methods.     

Dengue virus detection in Aedes aegypti larvae from southeastern Brazil

The transmission of dengue, the most important arthropod‐borne viral disease in Brazil, has been intensified over the past decades, along with the accompanying expansion and adaptation of its Aedes vectors. In the present study, we mapped dengue vectors in Ouro Preto and Ouro Branco, Minas Gerais, by installing ovitraps in 32 public schools. The traps were examined monthly between September, 2011 through July, 2012 and November, 2012 to April, 2013. The larvae were reared until the fourth stadium and identified according to species. The presence of dengue virus was detected by real time PCR and agarose gel electrophoresis. A total of 1,945 eggs was collected during the 17 months of the study. The Ovitrap Positivity Index (OPI) ranged from 0 to 28.13% and the Eggs Density Index (EDI) ranged from 0 to 59.9. The predominant species was Aedes aegypti, with 84.9% of the hatched larvae. Although the collection was low when compared to other ovitraps studies, vertical transmission could be detected. Of the 54 pools, dengue virus was detected in four Ae. aegypti pools.     

Oviposition by Aedes aegypti and Aedes albopictus: Influence of congeners and of oviposition site characteristics

We investigated the oviposition behavior of Ae. aegypti and Ae. albopictus. In particular we examined whether small‐scale site characteristics and the presence of conspecifics or congeners altered oviposition by these mosquitoes. Various combinations of females of the two species were allowed to oviposit inside cages among either vegetation (potted plants) or structural components (wood and concrete blocks). Numbers of eggs deposited per female were compared between species, sides, and treatments. Most significant differences between treatments and species involved differences between single species and mixed species treatments. Ae. aegypti deposited more eggs/female in the vegetation side than in the structure side whereas the opposite pattern was evident for Ae. albopictus. Ae. aegypti females had higher frequency of skip oviposition than Ae. albopictus. An average of 63% of the containers in the two‐species treatments contained eggs of both species, with more frequent joint occurrences observed in the treatment with three females of each species than in the treatments with one of each. Our results point to the existence of various interactions between gravid Ae. aegypti and Ae. albopictus females at or near the oviposition sites but further experimental work is necessary to fully characterize the interactions and their specific mechanisms.     

Behavioural responses of stable flies to cattle manure slurry associated odourants

Stable flies (Stomoxys calcitrans [Diptera: Muscidae] L.) are blood‐feeding synanthropic pests, which cause significant economic losses in livestock. Stable fly antennae contain olfactory sensilla responsive to host and host environment‐associated odours. Field observation indicated that the abundance of stable flies increased significantly in grasslands or crop fields when cattle manure slurry was applied. Major volatile compounds emanating from manure slurry were collected and identified. Behavioural responses of stable flies to those compounds were investigated in laboratory bioassays and field‐trapping studies. Results from olfactometer assays revealed that phenol, p‐cresol and m‐cresol were attractive to adult stable flies. When tested individually, attraction was higher with lower dosages. Stable flies were most attracted to blends of phenol and m‐cresol or p‐cresol. Traps with binary blend lures caught more stable flies in field trials as well.     

Field evaluations of disposable sticky lures for surveillance of Aedes aegypti (Stegomyia aegypti) and Culex quinquefasciatus in Jakarta

From December 1997 to April 1998, disposable sticky lures (1608 lure days) were trialled in homes in north Jakarta, Indonesia as surveillance tools for Aedes aegypti (Stegomyia aegypti) (Diptera: Culicidae) and Culex quinquefasciatus (Diptera: Culicidae), referenced to indoor resting adult collections (92 × 10 min). The lures collected 89.4% of the total of 1339 Ae. aegypti and 92.1% of the total of 1272 Cx. quinquefasciatus collected by all methods. Because there were no significant differences with respect to numbers collected in bedrooms, living rooms and kitchens, bedrooms were selected for subsequent trials for reasons of convenience. The main trials involved a replicated complete block design with L‐lysine and sodium carbonate. Lures without attractant or with four different dilutions of L‐lysine collected 3.4–8.5 times more Ae. aegypti and 4.2–8.1 times more Cx. quinquefasciatus than were collected by mouth aspirator. Lures with or without dilutions of sodium carbonate collected 2.7–5.0 times more Ae. aegypti and 1.8–4.2 times more Cx. quinquefasciatus than aspirator collections. The precision associated with catches of sticky lures was better than that for aspirator collections. Although olfactants generally improved the numbers of mosquitoes collected, the differences in catch between lures with and without attractants were usually non‐significant. Any deficit in catch may be offset by increasing the surveillance period to ≥30 days to detect all four dengue serotypes from infected mosquitoes.