Integrated Strategies for Aedes aegypti Control Applied to Individual Houses: An Approach to Mitigate Vectorial Arbovirus Transmission.

Aedes aegypti and Culex quinquefasciatus mosquitoes are vectors of different arboviruses that cause a large burden of disease in humans worldwide. A key step towards reducing the impact of arboviruses on humans can be achieved through integrated mosquito surveillance and control approaches. We carried out an integrated approach of mosquito surveillance and control actions to reduce populations of these insects along with a viral surveillance in a neighborhood of Recife (Northeastern Brazil) with high mosquito densities and arbovirus transmission. The actions were carried out in 40 different houses in the Nova Descoberta neighborhood. The area was divided into two groups, the control group using tools to monitor the mosquito density (1 OVT; 1 Double BR-ovt; monthly capture of alates) and the experimental group with control actions using surveillance tools in an intensified way (2 OVTs; 2 Double BR-ovts; fortnightly capture of alates; toxic baits). We evaluated the study's impact on the mosquito density via the Egg Density (ED) and Adult Density (AD) over a period of 12 cycles of 28 days each. The collected adult mosquitoes were processed via RT-qPCR for DENV, CHIKV and ZIKV and, subsequently, the Minimum Infection Rate (MIR) was calculated. After 12 cycles, we observed a 91% and 99% reduction in Aedes ED and AD in the monitored properties, as well as a 76% reduction in the AD of Cx. quinquefasciatus in the same properties. Moreover, we detected circulating arboviruses (DENV and ZIKV) in 19.52% of captured adult mosquitoes. We show that enhancing entomological surveillance tools can aid in the early detection of possible risk areas based on vector mosquito population numbers. Additionally, the detection of important arboviruses such as ZIKV and DENV raises awareness and allows for a better selection of risk areas and silent virus spread. It offers supplementary information for guiding emergency mosquito control measures in the target area. The goal is to minimize human-vector interactions and, subsequently, to lower the risk of transmitting circulating arboviruses.

Detection of Wuchereria bancrofti in the city of São Luís, state of Maranhão, Brazil: New incursion or persisting problem?

BACKGROUND: The elimination of lymphatic filariasis (LF) from Brazil by 2020 was not accomplished; however, this goal can be achieved in the upcoming years with the assistance of specific strategies. The surveillance of LF can be performed using molecular xenomonitoring (MX), a noninvasive method used to infer the presence of the parasite in the human population. Herein, São Luís (state of Maranhão) was the first city to be investigated to identify whether LF transmission in Brazil has been interrupted and if there were any new incursions. METHODOLOGY/PRINCIPAL FINDINGS: Mosquitoes were collected by aspiration at 901 points distributed among 11 neighborhoods in São Luís with records of patients with microfilaremia. Pools of engorged or gravid Culex quinquefasciatus females were evaluated by WbCx duplex PCR with endogenous control for mosquitoes and target for W. bancrofti for determining the vector infection rate. Among the 10,428 collected mosquitoes, the most abundant species were C. quinquefasciatus (85%) and Aedes aegypti (12%). Significantly larger numbers of mosquitoes were collected from the neighborhoods of Areinha and Coreia (p<0.05). MX performed using PCR validated 705 pools of engorged or gravid females, fifteen of which were positive for Wuchereria bancrofti in two neighborhoods. CONCLUSIONS: The high density of engorged C. quinquefasciatus females per home, inadequate sanitation, and detection of W. bancrofti-infected mosquitoes in the city of São Luís represent a warning of the possible upsurge of LF, a disease that is still neglected; this underscores the need for the ostensive monitoring of LF in Brazil.

Outbreak of autochthonous cases of malaria in coastal regions of Northeast Brazil: the diversity and spatial distribution of species of Anopheles.

BACKGROUND: Brazil has the fourth highest prevalence of malaria of all countries in the Americas, with an estimated 42 million people at risk of contracting this disease. Although most cases occur in the Amazon region, cases of an autochthonous nature have also been registered in the extra-Amazonian region where Anopheles aquasalis and An. albitarsis are the mosquito species of greatest epidemiological interest. In 2019, the municipality of Conde (state of Paraíba) experienced an epidemic of autochthonous cases of malaria. Here we present preliminary results of an entomological and case epidemiology investigation, in an attempt to correlate the diversity and spatial distribution of species of Anopheles with the autochthonous cases of this outbreak of malaria. METHODS: Case data were collected using case report forms made available by the Conde Municipal Health Department. The entomological survey was carried out from July to November 2019. The various methods of capture included the use of battery-powered aspirators, mouth aspirators, Shannon traps, BG-Sentinel traps (with and without dry ice) and CDC light traps. Captured mosquitoes were separated, packaged and sent to the laboratory for sexing and molecular identification of the various species of anophelines. The data were tabulated and analyzed using Microsoft Excel. Spatial analysis of the data was performed using ArcGis 10 software. RESULTS: In 2019, 20 autochthonous cases and one imported case of malaria caused by Plasmodium vivax were diagnosed, with three cases of relapses. A total of 3713 mosquitoes were collected, of which 3390 were culicines and 323 were anophelines. Nine species of genus Anopheles were identified, with the most abundant being An. aquasalis (38.9%), followed by An. minor (18.2%) and An. albitarsis (9.0%). Spatial analysis of the data showed that the area could be considered to be at risk of malaria cases and that there was a high prevalence of Anopheles. CONCLUSIONS: The results presented indicate that this extra-Amazonian region has an environment conducive to maintenance of the malaria transmission cycle owing to the wide diversity of Anopheles species. This environment in combination with the high influx of people from endemic areas to the study area provides a perfect setting for the occurrence and maintenance of malaria.

Double BR-OVT: a new trap model for collecting eggs and adult mosquitoes from Culex quinquefasciatus and Aedes spp.

The circulation of arboviruses throughout the world and the maintenance of lymphatic filariasis endemicity in tropical countries, combined with the lack of vaccines and specific treatments, highlight the importance of reducing the populations of mosquitoes involved in the transmission of these pathogens, Aedes aegypti and Culex quinquefasciatus. To contribute to the development of new strategies for monitoring and controlling these culicids, we evaluated the performance of the Double BR-OVT trap individually and in pairs, in the field. After 18 months, the Double BR-OVT traps captured a mean of 3.5 ± 7.4 and 1.8 ± 3.2 of Culex and Aedes /residence/cycle, respectively, in addition to 410 ± 588.3 Aedes eggs/residence/cycle. When installed in pairs, the Double BR-OVT traps collected three times more adult mosquitoes of C. quinquefasciatus (9.4 ± 8.3 Culex /residence/bimester) and two times more Aedes spp. (3 ± 3.2 Aedes /residence/bimester) in comparison with the traps installed individually (2.6 ± 7.1 and 1.5 ± 3.2 Culex and Aedes /residence/bimester, respectively) (p < 0.05). The Double BR-OVT trap has an exceptional advantage: it aggregates different functionalities into a single instrument, as this type of trap can concomitantly collect eggs and adult mosquitoes of C. quinquefasciatus and A. aegypti , a feature that makes it a potentially useful tool among the strategies for monitoring and controlling these mosquitoes.

Development of a molecular xenomonitoring protocol to assess filariasis transmission.

According to the World Health Organization, lymphatic filariasis (LF), a mosquito-borne neglected tropical disease (NTD), should be eliminated as a public health concern by the end of 2020. To this end, the goals of the Global Programme to Eliminate Lymphatic Filariasis (GPELF) include interrupting transmission through mass drug administration (MDA). After two decades, several countries have implemented MDA and are now ready to confirm whether transmission has been interrupted. The method for detecting the parasites in mosquito vectors known as xenomonitoring is a non-invasive tool for assessing the current transmission status of the filarial nematode Wuchereria bancrofti (which is responsible for 90% of cases) by their vectors. There are several methods available for detection of the worm in mosquito samples, such as dissection or polymerase chain reaction (PCR). However, most of these techniques still produce a considerable number of false-negative results. The present study describes a new duplex PCR protocol, which is an improvement on the traditional PCR methodology, enhanced by introducing the actin gene as an endogenous control gene. After adjusting the mosquito pool size, DNA extraction, and WbCx PCR duplex design, we achieved a reliable and sensitive molecular xenomonitoring protocol. This assay was able to eliminate 5% of false negative samples and detected less than one Wb larvae. This high sensitivity is particularly valuable after MDA, when prevalence declines. This new method could reduce the number of false-negative samples, which will enable us to improve our ability to generate accurate results and aid the monitoring strategies used by LF elimination programmes.

Infestation of an endemic arbovirus area by sympatric populations of Aedes aegypti and Aedes albopictus in Brazil.

BACKGROUND Aedes aegypti and Aedes albopictus are the most important arbovirus vectors in the world. OBJECTIVES This study aimed to investigate and compare the infestation pattern of these species in a neighbourhood of Recife, Brazil, endemic for arboviruses in 2005 (T1) and 2013 (T2). METHODS Infestation, distribution and relative abundance of these sympatric species were recorded by egg collection using a network of 59 sentinel ovitraps (s-ovt) at fixed sampling stations for 12 months in T1 and T2. FINDINGS A permanent occupation pattern was detected which was characterised by the presence of egg-laying females of one or both species with a high ovitrap positivity index (94.3 to 100%) throughout both years analysed. In terms of abundance, the total of eggs collected was lower (p < 0.005) in T2 (146,153) than in T1 (281,103), although ovitraps still displayed a high index of positivity. The spatial distribution showed the presence of both species in 65.1% of the 148 s-ovt assessed, while a smaller number of traps exclusively contained Ae. aegypti (22%) or Ae. albopictus (13.2%) eggs. MAIN CONCLUSIONS Our comparative analysis demonstrated the robustness of the spatial occupation and permanence of Ae. aegypti and Ae. albopictus populations in this endemic urban area.

Use of MALDI-TOF MS to identify the culturable midgut microbiota of laboratory and wild mosquitoes.

Mosquitoes are responsible for transmitting many pathogens to humans and Aedes aegypti, Aedes albopictus and Culex quinquefasciatus are important vectors in the world. The microbiota plays an important role in developmental studies that involve impacts on the biological cycle of mosquitoes and vector control strategies. In this study, the aim was to understand the environment plays in the microbiota culturable diversity of Aedes aegytpi, Aedes albopictus and Culex quinquefasciatus. Midgut of studied mosquitoes (laboratory-reared and wild) were dissected and analyzed by MALDI-TOF MS to identify the microbiota. Most of the bacteria identified in the microbiota of mosquitoes from the laboratory and field belong to the phylum Proteobacteria. We reported on the microbial diversity among the mosquito species studied where Cx. quinquefasciatus and Ae. albopictus show greater bacterial similarity. The genus Rahnella was present in all mosquito species studied, both in those from the laboratory and those from the wild. Bacillus, Ewingella, Microccocus, Klebsiella and Pantoea are genera was predominant among the mosquitoes studied. The difference of microbiota diversity between mosquitoes laboratory-reared and wild shows that the environment plays an important role in the acquisition of bacteria, mainly in Ae. aegypti and Cx. quinquefasciatus.

Zika virus detection, isolation and genome sequencing through Culicidae sampling during the epidemic in Vitória, Espírito Santo, Brazil.

BACKGROUND: Zika virus (ZIKV) has been isolated from many mosquito species in nature, but it is believed that the main vectors in urban environments are species of the genus Aedes. Here, we detected and isolated ZIKV in samples from Aedes aegypti, Aedes taeniorhynchus and Culex quinquefasciatus, collected during the Zika epidemic in Vitória, southeast Brazil. Using quantitative real-time polymerase chain reaction, ZIKV detection was performed in mosquito samples collected from February to April 2016. RESULTS: Overall, six pools of mosquitoes were positive for ZIKV: four of Cx. quinquefasciatus, one of Ae. aegypti and one of Ae. taeniorhynchus. Their genomes were sequenced. CONCLUSIONS: These results support and strengthen the hypothesis that other mosquito species can also be involved in ZIKV transmission.

Long-term exposure of Aedes aegypti to Bacillus thuringiensis svar. israelensis did not involve altered susceptibility to this microbial larvicide or to other control agents.

BACKGROUND: Bacillus thuringiensis svar. israelensis (Bti) is an effective and safe biolarvicide to control Aedes aegypti. Its mode of action based on four protoxins disfavors resistance; however, control in endemic areas that display high mosquito infestation throughout the year requires continuous larvicide applications, which imposes a strong selection pressure. Therefore, this study aimed to investigate the effects of an intensive Bti exposure on an Ae. aegypti strain (RecBti), regarding its susceptibility to Bti and two of its protoxins tested individually, to other control agents temephos and diflubenzuron, and its profile of detoxifying enzymes. METHODS: The RecBti strain was established using a large egg sample (10,000) from Recife city (Brazil) and more than 290,000 larvae were subjected to Bti throughout 30 generations. Larvae susceptibility to larvicides and the activity of detoxifying enzymes were determined by bioassays and catalytic assays, respectively. The Rockefeller strain was the reference used for these evaluations. RESULTS: Bti exposure yielded an average of 74% mortality at each generation. Larvae assessed in seven time points throughout the 30 generations were susceptible to Bti crystal (resistance ratio RR ≤ 2.8) and to its individual toxins Cry11Aa and Cry4Ba (RR ≤ 4.1). Early signs of altered susceptibility to Cry11Aa were detected in the last evaluations, suggesting that this toxin was a marker of the selection pressure imposed. RecBti larvae were also susceptible (RR ≤ 1.6) to the other control agents, temephos and diflubenzuron. The activity of the detoxifying enzymes α- and ß-esterases, glutathione-S-transferases and mixed-function oxidases was classified as unaltered in larvae from two generations (F19 and F25), except for a ß-esterases increase in F25. CONCLUSIONS: Prolonged exposure of Ae. aegypti larvae to Bti did not evolve into resistance to the crystal, and no cross-resistance with temephos and diflubenzuron were recorded, which supports their sustainable use with Bti for integrated control practices. The unaltered activity of most detoxifying enzymes suggests that they might not play a major role in the metabolism of Bti toxins, therefore resistance by this mechanism is unlikely to occur. This study also highlights the need to establish suitable criteria to classify the status of larval susceptibility/resistance.

Culicidae fauna (Diptera: Culicidae) survey in urban, ecotonal and forested areas, from the Moreno municipality - Pernambuco State, Brazil.

INTRODUCTION: Mosquitoes are vectors of several pathogens in tropical regions. However, information on Culicidae in the northeast region of Brazil is scarce despite many municipalities having environmental conditions favorable for these insects. METHODS: We collected mosquitoes monthly for one year from urban, ecotonal, and ecological reserve areas in Moreno, Brazil, by using CDC Miniature light traps, aspiration, ovitraps, and liquid suction. RESULTS: We collected 1,401 insects in the Culicidae family. Fifteen species of seven genera of Culicidae were identified and five specimens were identified only at the genus level. CONCLUSIONS: There is a wide diversity of mosquito species in the sampled areas, some of which are important for human health.

Culicidae fauna (Diptera: Culicidae) survey in urban, ecotonal and forested areas, from the Moreno municipality - Pernambuco State, Brazil

Abstract INTRODUCTION: Mosquitoes are vectors of several pathogens in tropical regions. However, information on Culicidae in the northeast region of Brazil is scarce despite many municipalities having environmental conditions favorable for these insects. METHODS: We collected mosquitoes monthly for one year from urban, ecotonal, and ecological reserve areas in Moreno, Brazil, by using CDC Miniature light traps, aspiration, ovitraps, and liquid suction. RESULTS: We collected 1,401 insects in the Culicidae family. Fifteen species of seven genera of Culicidae were identified and five specimens were identified only at the genus level. CONCLUSIONS: There is a wide diversity of mosquito species in the sampled areas, some of which are important for human health.

Field evaluation of sticky BR-OVT traps to collect culicids eggs and adult mosquitoes inside houses.

INTRODUCTION: Culex quinquefasciatus is a mosquito of importance to public health, as it represents a real and/or potential risk for the transmission of pathogens to humans, such as some arthropod-borne viruses and nematodes that cause filariasis. In Brazil, three municipalities in the Metropolitan Region of Recife (RMR) that are endemic for lymphatic filariasis conducted control actions targeting this vector. With the goal of contributing novel C. quinquefasciatus collection strategies, a sticky trap capable of collecting eggs and imprisoning mosquitoes was investigated. METHODS: To evaluate the performance of the sticky BR-OVT trap, tests were carried out in the neighborhoods of Caixa d'Água and Passarinho (Olinda-PE-Brasil) between August 2011 and June 2012. Sixty traps were installed in the indoor areas of residences in the two districts. RESULTS: During the 11-month study, 0.52 [standard deviation (SD) = 1.52] Culex egg rafts, 2.16 (SD = 4.78) C. quinquefasciatus/trap/month, and 0.55 (SD = 1.28) Aedes/trap/month were caught. Female specimens predominated the traps (59% of C. quinquefasciatus and 96% of Aedes spp.). CONCLUSIONS: The results demonstrated that the sticky BR-OVT trap is a useful tool for the collection of adult culicids of medical importance and offers an innovative way to collect C. quinquefasciatus eggs and adults in a single trap.

Field evaluation of sticky BR-OVT traps to collect culicids eggs and adult mosquitoes inside houses

Abstract INTRODUCTION: Culex quinquefasciatus is a mosquito of importance to public health, as it represents a real and/or potential risk for the transmission of pathogens to humans, such as some arthropod-borne viruses and nematodes that cause filariasis. In Brazil, three municipalities in the Metropolitan Region of Recife (RMR) that are endemic for lymphatic filariasis conducted control actions targeting this vector. With the goal of contributing novel C. quinquefasciatus collection strategies, a sticky trap capable of collecting eggs and imprisoning mosquitoes was investigated. METHODS: To evaluate the performance of the sticky BR-OVT trap, tests were carried out in the neighborhoods of Caixa d'Água and Passarinho (Olinda-PE-Brasil) between August 2011 and June 2012. Sixty traps were installed in the indoor areas of residences in the two districts. RESULTS: During the 11-month study, 0.52 [standard deviation (SD) = 1.52] Culex egg rafts, 2.16 (SD = 4.78) C. quinquefasciatus/trap/month, and 0.55 (SD = 1.28) Aedes/trap/month were caught. Female specimens predominated the traps (59% of C. quinquefasciatus and 96% of Aedes spp.). CONCLUSIONS The results demonstrated that the sticky BR-OVT trap is a useful tool for the collection of adult culicids of medical importance and offers an innovative way to collect C. quinquefasciatus eggs and adults in a single trap.

Insecticide susceptibility of Aedes albopictus and Ae. aegypti from Brazil and the Swiss-Italian border region.

BACKGROUND: Aedes aegypti and Ae. albopictus are two highly invasive mosquito species, both vectors of several viruses, including dengue, chikungunya and Zika. While Ae. aegypti is the primary vector in the tropics and sub-tropics, Ae. albopictus is increasingly under the public health watch as it has been implicated in arbovirus-transmission in more temperate regions, including continental Europe. Vector control using insecticides is the pillar of most control programmes; hence development of insecticide resistance is of great concern. As part of a Brazilian-Swiss Joint Research Programme we set out to assess whether there are any signs of existing or incipient insecticide resistance primarily against the larvicide Bacillus thuringiensis svar. israelensis (Bti), but also against currently applied and potentially alternative insecticides in our areas, Recife (Brazil) and the Swiss-Italian border region. METHODS: Following World Health Organization guidelines, dose-response curves for a range of insecticides were established for both colonized and field caught Ae. aegypti and Ae. albopictus. The larvicides included Bti, two of its toxins, Cry11Aa and Cry4Ba, Lysinibacillus sphaericus, Vectomax CG®, a formulated combination of Bti and L. sphaericus, and diflubenzuron. In addition to the larvicides, the Swiss-Italian Ae. albopictus populations were also tested against five adulticides (bendiocarb, dichlorodiphenyltrichloroethane, malathion, permethrin and λ-cyhalothrin). RESULTS: Showing a similar dose-response, all mosquito populations were fully susceptible to the larvicides tested and, in particular, to Bti which is currently used both in Brazil and Switzerland. In addition, there were no signs of incipient resistance against Bti as larvae were equally susceptible to the individual toxins, Cry11Aa and Cry4Ba. The field-caught Swiss-Italian populations were susceptible to the adulticides tested but DDT mortality rates showed signs of reduced susceptibility. CONCLUSIONS: The insecticides currently used for mosquito control in Switzerland and Brazil are still effective against the target populations. The present study provides an important reference as relatively few insecticide susceptibility surveys have been carried out with Ae. albopictus.

A new allele conferring resistance to Lysinibacillus sphaericus is detected in low frequency in Culex quinquefasciatus field populations.

BACKGROUND: The Cqm1 α-glucosidase of Culex quinquefasciatus larvae acts as the midgut receptor for the binary toxin of the biolarvicide Lysinibacillus sphaericus. Mutations within the cqm1 gene can code for aberrant polypeptides that can no longer be properly expressed or bind to the toxin, leading to insect resistance. The cqm1 REC and cqm1 REC-2 alleles were identified in a laboratory selected colony and both displayed mutations that lead to equivalent phenotypes of refractoriness to L. sphaericus. cqm1 REC was first identified as the major resistance allele in this colony but it was subsequently replaced by cqm1 REC-2 , suggesting the better adaptive features of the second allele. The major aim of this study was to evaluate the occurrence of cqm1 REC-2 and track its origin in field populations where cqm1 REC was previously identified. METHODS: The screening of the cqm1 REC-2 allele was based on more than 2000 C. quinquefasciatus larvae from five localities in the city of Recife, Brazil and used a multiplex PCR assay that is also able to identify cqm1 REC . Full-length sequencing of the cqm1 REC-2 and selected cqm1 samples was performed to identify further polymorphisms between these alleles. RESULTS: The cqm1 REC-2 allele was found in field samples, specifically in two heterozygous individuals from a single locality with an overall frequency and distribution much lower than that observed for cqm1 REC . The full-length sequences from these two cqm1 REC-2 copies were almost identical to the cqm1 REC-2 derived from the resistant colony but displayed more than 30 SNPs when compared with cqm1 and cqm1 REC . The cqm1 REC and cqm1 REC-2 resistant alleles were found to be associated with two distinct sets of wild-type cqm1 variants found in field populations. CONCLUSIONS: The cqm1 REC-2 allele occurs in populations in Recife and was probably already present in the samples used to establish the laboratory resistant colony. The data generated indicates that cqm1 REC-2 can be selected in field populations, although its low frequency and distribution in Recife suggest that cqm1 REC-2 presents a lower risk of selection compared to cqm1 REC .

Vector competence of the Aedes aegypti population from Santiago Island, Cape Verde, to different serotypes of dengue virus.

BACKGROUND: Dengue is an arboviral disease caused by dengue virus (DENV), whose main vectors are the mosquitoes Aedes aegypti and Aedes albopictus. A. aegypti is the only DENV vector in Cape Verde, an African country that suffered its first outbreak of dengue in 2009. However, little is known about the variation in the level of vector competence of this mosquito population to the different DENV serotypes. This study aimed to evaluate the vector competence of A. aegypti from the island of Santiago, Cape Verde, to four DENV serotypes and to detect DENV vertical transmission. METHODS: Mosquitoes were fed on blood containing DENV serotypes and were dissected at 7, 14 and 21 days post-infection (dpi) to detect the virus in the midgut, head and salivary glands (SG) using RT-PCR. Additionally, the number of copies of viral RNA present in the SG was determined by qRT-PCR. Furthermore, eggs were collected in the field and adult mosquitoes obtained were analyzed by RT-PCR and the platelia dengue NS1 antigen kit to detect transovarial transmission. RESULTS: High rates of SG infection were observed for DENV-2 and DENV-3 whereas for DENV-1, viral RNA was only detected in the midgut and head. DENV-4 did not spread to the head or SG, maintaining the infection only in the midgut. The number of viral RNA copies in the SG did not vary significantly between DENV-2 and DENV-3 or among the different periods of incubation and the various titers of DENV tested. With respect to DENV surveillance in mosquitoes obtained from the eggs collected in the field, no samples were positive. CONCLUSION: Although no DENV positive samples were collected from the field in 2014, it is important to highlight that the A. aegypti population from Santiago Islands exhibited different degrees of susceptibility to DENV serotypes. This population showed a high vector competence for DENV-2 and DENV-3 strains and a low susceptibility to DENV-1 and DENV-4. Viral RNA copies in the SG remained constant for at least 21 dpi, which may enhance the vector capacity of A. aegypti and suggests the presence of a mechanism modulating virus replication in the SG.

Characterization of the spatial and temporal dynamics of the dengue vector population established in urban areas of Fernando de Noronha, a Brazilian oceanic island.

Aedes aegypti has played a major role in the dramatic expansion of dengue worldwide. The failure of control programs in reducing the rhythm of global dengue expansion through vector control suggests the need for studies to support more appropriated control strategies. We report here the results of a longitudinal study on Ae. aegypti population dynamics through continuous egg sampling aiming to characterize the infestation of urban areas of a Brazilian oceanic island, Fernando de Noronha. The spatial and temporal distribution of the dengue vector population in urban areas of the island was described using a monitoring system (SMCP-Aedes) based on a 103-trap network for Aedes egg sampling, using GIS and spatial statistics analysis tools. Mean egg densities were estimated over a 29-month period starting in 2011 and producing monthly maps of mosquito abundance. The system detected continuous Ae. aegypti oviposition in most traps. The high global positive ovitrap index (POI=83.7% of 2815 events) indicated the frequent presence of blood-fed-egg laying females at every sampling station. Egg density (eggs/ovitrap/month) reached peak values of 297.3 (0 - 2020) in May and 295 (0 - 2140) in August 2012. The presence of a stable Ae. aegypti population established throughout the inhabited areas of the island was demonstrated. A strong association between egg abundance and rainfall with a 2-month lag was observed, which combined with a first-order autocorrelation observed in the series of egg counts can provide an important forecasting tool. This first description of the characteristics of the island infestation by the dengue vector provides baseline information to analyze relationships between the spatial distribution of the vector and dengue cases, and to the development of integrated vector control strategies.

Effect of Moringa oleifera lectins on survival and enzyme activities of Aedes aegypti larvae susceptible and resistant to organophosphate.

The indiscriminate use of synthetic insecticides to control Aedes aegypti has led to emergence of resistant populations. Moringa oleifera seeds contain the lectins WSMoL and cMoL. WSMoL has larvicidal activity on fourth-stage of A. aegypti organophosphate-susceptible larvae (Rockefeller L4). This study reports on the effects of cMoL on the survival of Rockefeller L4 as well as of WSMoL and cMoL on L4 from an organophosphate-resistant population (Rec-R). The effects of lectins on digestive (amylase, trypsin, and protease) and detoxifying (superoxide dismutase (SOD), α- and ß-esterases) enzymes from larvae were also determined. cMoL (0.1-0.8 mg/ml) did not kill Rockefeller L4 as well as WSMoL and cMoL (0.1-0.8 mg/ml) were not larvicidal for Rec-R L4. WSMoL stimulated protease, trypsin-like, and α-amylase from Rockefeller L4 while cMoL inhibited these enzymes. WSMoL had no effect on trypsin-like activity from Rec-R L4 but inhibited protease and α-amylase. Among digestive enzymes of Rec-R L4, cMoL inhibited only trypsin-like activity. cMoL inhibited SOD activities from Rockefeller and Rec-R L4 in a higher level than WSMoL while ß-esterase from Rockefeller L4 was more inhibited by WSMoL. The lectins promoted low stimulation or inhibition of α-esterase activities from both populations. In conclusion, Rockefeller and Rec-R larvae were distinctly affected by M. oleifera lectins, and larvicidal mechanism of WSMoL on Rockefeller L4 may involve deregulation of digestive enzymes. cMoL interfered mainly on SOD activity and thus it can be investigated as a synergistic agent for controlling populations whose resistance is linked to an increased detoxifying process mediated by this enzyme.

Susceptibility status of Culex quinquefasciatus (Diptera: Culicidae) populations to the chemical insecticide temephos in Pernambuco, Brazil.

BACKGROUND: Culex quinquefasciatus is the vector of many agents of human diseases, including Wuchereria bancrofti, the parasite that causes bancroftian filariasis, an endemic disease in Pernambuco State, Brazil. Although temephos is not currently used to control C. quinquefasciatus, the species might be under a selection process from incidental exposure to this compound. This study aimed to evaluate the susceptibility status of C. quinquefasciatus to temephos, using bioassays, and to investigate its putative resistance mechanisms through biochemical assays and screening of the G119S mutation in the acetylcholinesterase gene, which is associated with organophosphate resistance, carried out by PCR and sequencing. RESULTS: The results showed that only mosquitoes from Santa Cruz do Capibaribe (SC) had an alteration in their susceptibility status (RR = 7.2-fold), while the other populations were all susceptible to the insecticide. Biochemical assays showed increased activity for all esterases in SC, as well as evidence of acetylcholinesterase insensitivity. The G119S mutation was detected in this population with a frequency of 0.11, but it was not found in the remaining populations. CONCLUSION: These data show that mechanisms of temephos resistance have been selected in natural C. quinquefasciatus populations from Pernambuco, which could undermine future control actions.

The susceptibility of Aedes aegypti populations displaying temephos resistance to Bacillus thuringiensis israelensis: a basis for management.

BACKGROUND: Aedes aegypti is the vector of dengue virus, and its control is essential to prevent disease transmission. Among the agents available to control this species, biolarvicides based on Bacillus thuringiensis serovar israelensis (Bti) are an effective alternative to replace the organophosphate temephos for controlling populations that display resistance to this insecticide. The major goal of this study was to determine the baseline susceptibility of Brazilian Ae. aegypti populations to Bti, taking into account their background in terms of larvicide exposure, status of temephos resistance and the level of activity of detoxifying enzymes involved in metabolic resistance to insecticides. METHODS: Population samples were established under insectarium conditions. Larval susceptibility to temephos and Bti was evaluated through bioassays and lethal concentrations of these compounds were determined. Biochemical assays were performed to determine the specific activity of five detoxifying enzymes in these samples. RESULTS: Fourteen populations were characterized and, except for one case, all displayed resistance to temephos. Most populations were classified as highly resistant. The populations also showed increased activity of one or more detoxifying enzymes (glutathione-S-transferases, esterases and mixed function oxidases), regardless of their temephos resistance status. All populations analyzed were susceptible to Bti, and the lethal concentrations were similar to those detected in two laboratory susceptible colonies. The response to Bti showed little variation. A maximum resistance ratio of 2.1 was observed in two untreated populations, while in two Bti-treated populations, the maximum resistance ratio was 1.9. No positive correlation was found between temephos resistance, increased activity of detoxifying enzymes, and susceptibility to Bti. CONCLUSIONS: Data from this study show that all populations were susceptible to Bti, including twelve untreated and two treated populations that had been exposed to this agent for more than ten years. The temephos resistance and increased activity of detoxifying enzymes observed in thirteen populations was not correlated with changes in susceptibility to Bti. Our data show a lack of cross-resistance between these two compounds; thus, Bti can be used in an integrated control program to fight Ae. aegypti and counteract the temephos resistance that was found among all populations analyzed.