Mosquito Culex Quinquefasciatus Research Articles

Histological validation of in-vivo larvicidal efficacy of marine Streptomyces sp. RD06 secondary metabolites against filariasis causing Culex quinquefasciatus and statistical media optimization for larvicidal derivatives production

Mosquito-borne disease pandemics, such as the Zika virus and chikungunya, have escalated cognizance of how critical it is to implement proficient mosquito vector control measures. The prevention of Culicidae is becoming more difficult these days because of the expeditious imminence of synthetic pesticide resistance and the universal expansion of tremendously invasive mosquito vectors. The present study highlights the insecticidal and larvicidal efficacy of the prospective novel actinobacterium derived from the marine Streptomyces sp. RD06 secondary metabolites against Culex quinquefasciatus mosquito. The pupicidal activity of Streptomyces sp. RD06 showed LC50=199.22±11.54 and LC90= 591.84 ±55.41 against the pupa. The purified bioactive metabolites 1, 2-Benzenedicarboxylic acid, diheptyl ester from Streptomyces sp. RD06 exhibited an LC50 value of 154.13±10.50 and an LC90 value of 642.84±74.61 tested against Cx. quinquefasciatus larvae. The Streptomyces sp. RD06 secondary metabolites exhibited 100 % non-hatchability at 62.5 ppm, and 82 % of hatchability was observed at 250 ppm. In addition, media optimization showed that the highest biomass production was attained at a temperature of 41.44°C, pH 9.23, nitrogen source 11.43 mg/ml, and carbon source 150 mg/ml. Compared to control larvae, the histology and confocal microscopy results showed destruction to the anal gill, lumen content, and epithelial layer residues in the treated larvae. Utilizing an eco-friendly method, these alternative inventive insecticidal derivatives from Streptomyces sp. RD06 eradicates Culex quinquefasciatus. This study highlights the promising potential of these Streptomyces sp. RD06 secondary metabolites to develop affordable and efficacious mosquito larvicides to replace synthetic insecticides in the future.

Antibodies to Aedes aegypti D7L salivary proteins as a new serological tool to estimate human exposure to Aedes mosquitoes.

Aedes spp. are the most prolific mosquito vectors in the world. Found on every continent, they can effectively transmit various arboviruses, including the dengue virus which continues to cause outbreaks worldwide and is spreading into previously non-endemic areas. The lack of widely available dengue vaccines accentuates the importance of targeted vector control strategies to reduce the dengue burden. High-throughput tools to estimate human-mosquito contact and evaluate vector control interventions are lacking. We propose a novel serological tool that allows rapid screening of human cohorts for exposure to potentially infectious mosquitoes. We tested 563 serum samples from a longitudinal pediatric cohort study previously conducted in Cambodia. Children enrolled in the study were dengue-naive at baseline and were followed biannually for dengue incidence for two years. We used Western blotting and enzyme-linked immunosorbent assays to identify immunogenic Aedes aegypti salivary proteins and measure total anti-Ae. aegypti IgG. We found a correlation (rs=0.86) between IgG responses against AeD7L1 and AeD7L2 recombinant proteins and those to whole salivary gland homogenate. We observed seasonal fluctuations of AeD7L1+2 IgG responses and no cross-reactivity with Culex quinquefasciatus and Anopheles dirus mosquitoes. The baseline median AeD7L1+2 IgG responses for young children were higher in those who developed asymptomatic versus symptomatic dengue. The IgG response against AeD7L1+2 recombinant proteins is a highly sensitive and Aedes specific marker of human exposure to Aedes bites that can facilitate standardization of future serosurveys and epidemiological studies by its ability to provide a robust estimation of human-mosquito contact in a high-throughput fashion.

Terrestrial subsidies and light affect an aquatic micro‐ecosystem in unexpected ways

Abstract The effects of resource subsidies on a recipient ecosystem depends on a variety of factors. Firstly, increasing the supply of limiting nutrients tends to increase the accumulation of biomass across different compartments of the recipient food web. Secondly, in the case of aquatic microorganisms, this accumulation may also be contingent on inputs of energy through light, mediating interactions between heterotrophic bacteria and autotrophic algae. Thirdly, macroscopic consumers feed upon microorganisms and recycle nutrients through excretion and egestion, profoundly affecting nutrient cycling. In order to examine the context‐dependency of nutrient cycling in aquatic systems, we performed an experiment in mesocosms where we followed the responses of dissolved nutrients, microorganisms, and larvae of the cosmopolitan mosquito Culex quinquefasciatus (Diptera: Culicidae) to two terrestrial subsidies differing in carbon (C):nutrient, in combination with two light intensities. We found that the interactions amongst bacteria and algae themselves remained generally unimportant after accounting for treatment effects, and that light did mediate some of the effects of nutrient subsidies on microorganisms. We also found that the presence of mosquito larvae was associated with altered concentrations of nitrogen (N) in some treatments, ultimately benefitting algae. Finally, we found that low C:nutrient subsidies generally increased the development rate of mosquito larvae, and that increased light exposure led to higher mosquito survival. Overall, these patterns may be explained by strong nitrogen limitation in our system, causing both group of microorganisms to respond similarly to variations in nutrient supply without direct covariation, and to the key role of N recycling by mosquito larvae in sustaining algal densities in exposed treatments. The context‐dependency in our study may reflect elemental or molecular constraints other than phosphorus. Our experiment thus calls for greater synthesis amongst ecological theories.

Infection, dissemination, and transmission of lumpy skin disease virus in Aedes aegypti (Linnaeus), Culex tritaeniorhynchus (Giles), and Culex quinquefasciatus (Say) mosquitoes

Lumpy skin disease virus (LSDV) is a transboundary viral disease in cattle and water buffaloes. Although this Poxvirus is supposedly transmitted by mechanical vectors, only a few studies have investigated the role of local vectors in the transmission of LSDV. This study examined the infection, dissemination, and transmission rates of LSDV in Aedes aegypti, Culex tritaeniorhynchus, and Culex quinquefasciatus following artificial membrane feeding of 102.7, 103.7, 104.7 TCID50/mL LSDV in sheep blood. The results demonstrated that these mosquito species were susceptible to LSDV, with Cx tritaeniorhynchus exhibiting significantly different characteristics from Ae. aegypti and Cx. quinquefasciatus. These three mosquito species were susceptible to LSDV. Ae. aegypti showed it as early as 2 days post-infection (dpi), indicating swift dissemination in this particular species. The extrinsic incubation period (EIP) of LSDV in Cx. tritaeniorhynchus and Cx. quinquefasciatus was 8 and 14 dpi, respectively. Ingestion of different viral titers in blood did not affect the infection, dissemination, or transmission rates of Cx. tritaeniorhynchus and Cx. quinquefasciatus. All rates remained consistently high at 8–14 dpi for Cx. tritaeniorhynchus. In all three species, LSDV remained detectable until 14 dpi. The present findings indicate that, Ae. aegypti, Cx. tritaeniorhynchus, and Cx. quinquefasciatus may act as vectors during the LSDV outbreak; their involvement may extend beyond being solely mechanical vectors.

Application of computer vision and deep learning models to automatically classify medically important mosquitoes in North Borneo, Malaysia.

Mosquito-borne diseases have emerged in North Borneo in Malaysia due to rapid changes in the forest landscape, and mosquito surveillance is key to understanding disease transmission. However, surveillance programmes involving sampling and taxonomic identification require well-trained personnel, are time-consuming and labour-intensive. In this study, we aim to use a deep leaning model (DL) to develop an application capable of automatically detecting mosquito vectors collected from urban and suburban areas in North Borneo, Malaysia. Specifically, a DL model called MobileNetV2 was developed using a total of 4880 images of Aedes aegypti, Aedes albopictus and Culex quinquefasciatus mosquitoes, which are widely distributed in Malaysia. More importantly, the model was deployed as an application that can be used in the field. The model was fine-tuned with hyperparameters of learning rate 0.0001, 0.0005, 0.001, 0.01 and the performance of the model was tested for accuracy, precision, recall and F1 score. Inference time was also considered during development to assess the feasibility of the model as an app in the real world. The model showed an accuracy of at least 97%, a precision of 96% and a recall of 97% on the test set. When used as an app in the field to detect mosquitoes with the elements of different background environments, the model was able to achieve an accuracy of 76% with an inference time of 47.33 ms. Our result demonstrates the practicality of computer vision and DL in the real world of vector and pest surveillance programmes. In the future, more image data and robust DL architecture can be explored to improve the prediction result.

The infection kinetics and transmission potential of two Guaico Culex viruses in Culex quinquefasciatus mosquitoes

Guaico Culex virus (GCXV) is a newly identified segmented Jingmenvirus from Culex spp. mosquitoes in Central and South America. The genome of GCXV is composed of four or five single-stranded positive RNA segments. However, the infection kinetics and transmission capability of GCXV in mosquitoes remain unknown. In this study, we used reverse genetics to rescue two GCXVs (4S and 5S) that contained four and five RNA segments, respectively, in C6/36 ​cells. Further in vitro characterization revealed that the two GCXVs exhibited comparable replication kinetics, protein expression and viral titers. Importantly, GCXV RNAs were detected in the bodies, salivary glands, midguts and ovaries of Culex quinquefasciatus at 4–10 days after oral infection. In addition, two GCXVs can colonize Cx. quinquefasciatus eggs, resulting in positive rates of 15%–35% for the second gonotrophic cycle. In conclusion, our results demonstrated that GCXVs with four or five RNA segments can be detected in Cx. quinquefasciatus eggs during the first and second gonotrophic cycles after oral infection.

Biolarvicide activity of extract and essential oil of culantro (Eryngium foetidum L.) on Culex quinquefasciatus larvae in silico and in vitro


 
 
 
 Background: Filariasis, a priority tropical disease in Indonesia, is transmitted by the Culex quinquefasciatus mosquito. The use of conventional synthetic larvicides can lead to environmental and health issues, including poisoning and resistance in target insect populations.
 Objective: This study explores the utilization of biolarvicides derived from culantro (Eryngium foetidum) to mitigate these adverse efects, focusing on analyzing the components and evaluating the larvicidal eficacy of both the extract and essential oil of E. foetidum.
 Method: This study employed a molecular docking approach to examine in silico biolarvicidal activity against the odorant binding protein (OBP) receptor and conducted in vitro experiments on Cx. quinquefasciatus larvae using varying concentrations of E. foetidum extract (100, 250, and 500 ppm) and essential oil (10, 50, and 100 ppm).
 Results: The in silico study identified hynokiflavone and longifenaldehyde as the compounds with the most potent activity, demonstrating binding a inities of -10.2 and -9.5 kcal/mol, respectively. The in vitro assays revealed that the E. foetidum extract achieved 75% larval mortality at an LC50 of 78.59 ppm, while the essential oil resulted in 88% mortality with an LC50 of 10.13 ppm.
 Conclusion:The extract and essential oil of E. foetidum exhibit significant biolarvicidal activity against Culex quinquefasciatus, offering promising plant-based alternatives to traditional larvicides, with implications for safer and more sustainable vector control strategies.
 
 
 

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.

Postintervention Immunological and Entomological Survey of Lymphatic Filariasis in the City of Olinda, Brazil, 2015-2016.

Lymphatic filariasis (LF) is a leading cause of disability due to infectious disease worldwide. The Recife Metropolitan Region (RMR) is the only remaining focus of LF in Brazil, where the parasite Wuchereria bancrofti is transmitted solely by the mosquito Culex quinquefasciatus. This study reports the results of transmission assessment surveys and molecular xenomonitoring in the city of Olinda, RMR, after nearly 15 years (2015-2016) of interventions for LF elimination. Participants were screened for W. bancrofti antigen via immunochromatographic card tests (ICT) in: 1) door-to-door surveys conducted for all children aged 5-7 years from 4 out of 17 intervention areas treated with at least five annual doses of mass drug administration (MDA), and 2) a two-stage cluster sampling survey of residents aged 5 years and older in non-MDA areas. Mosquitoes were collected via handheld aspirators in four MDA areas, differentiated by species, sex, and physiological status, pooled into groups of up to 10 blood-fed, semigravid, and gravid mosquitoes, and screened for W. bancrofti infection by real-time quantitative polymerase chain reaction (RT-qPCR). All 1,170 children from MDA areas and the entire population sample of 990 residents in non-MDA areas were ICT negative. In MDA areas, a total of 3,152 female Cx. quinquefasciatus mosquitoes in 277 households (range, 0-296 mosquitoes per house) were collected via aspiration. RT-qPCR of 233 pools of mosquitos were negative for W. bancrofti RNA; an independent reference laboratory confirmed these results. These results provide evidence that LF transmission has been halted in this setting.

Environmental and geographical factors influence the occurrence and abundance of the southern house mosquito, Culex quinquefasciatus, in Hawai‘i

Hawaiian honeycreepers, a group of endemic Hawaiian forest birds, are being threatened by avian malaria, a non-native disease that is driving honeycreepers populations to extinction. Avian malaria is caused by the parasite Plasmodium relictum, which is transmitted by the invasive mosquito Culex quinquefasciatus. Environmental and geographical factors play an important role in shaping mosquito-borne disease transmission dynamics through their influence on the distribution and abundance of mosquitoes. We assessed the effects of environmental (temperature, precipitation), geographic (site, elevation, distance to anthropogenic features), and trap type (CDC light trap, CDC gravid trap) factors on mosquito occurrence and abundance. Occurrence was analyzed using classification and regression tree models (CART) and generalized linear models (GLM); abundance (count data) was analyzed using generalized linear mixed models (GLMMs). Models predicted highest mosquito occurrence at mid-elevation sites and between July and November. Occurrence increased with temperature and precipitation up to 580 mm. For abundance, the best model was a zero-inflated negative-binomial model that indicated higher abundance of mosquitoes at mid-elevation sites and peak abundance between August and October. Estimation of occurrence and abundance as well as understanding the factors that influence them are key for mosquito control, which may reduce the risk of forest bird extinction.

Structure of the Lysinibacillus sphaericus Tpp49Aa1 pesticidal protein elucidated from natural crystals using MHz-SFX.

The Lysinibacillus sphaericus proteins Tpp49Aa1 and Cry48Aa1 can together act as a toxin toward the mosquito Culex quinquefasciatus and have potential use in biocontrol. Given that proteins with sequence homology to the individual proteins can have activity alone against other insect species, the structure of Tpp49Aa1 was solved in order to understand this protein more fully and inform the design of improved biopesticides. Tpp49Aa1 is naturally expressed as a crystalline inclusion within the host bacterium, and MHz serial femtosecond crystallography using the novel nanofocus option at an X-ray free electron laser allowed rapid and high-quality data collection to determine the structure of Tpp49Aa1 at 1.62 Å resolution. This revealed the packing of Tpp49Aa1 within these natural nanocrystals as a homodimer with a large intermolecular interface. Complementary experiments conducted at varied pH also enabled investigation of the early structural events leading up to the dissolution of natural Tpp49Aa1 crystals-a crucial step in its mechanism of action. To better understand the cooperation between the two proteins, assays were performed on a range of different mosquito cell lines using both individual proteins and mixtures of the two. Finally, bioassays demonstrated Tpp49Aa1/Cry48Aa1 susceptibility of Anopheles stephensi, Aedes albopictus, and Culex tarsalis larvae-substantially increasing the potential use of this binary toxin in mosquito control.

Bioactive Potentials of Petunia Integrifolia (Hook.) Schinz & Thell. Against Culex Quinquefasciatus Larvae, Artemia Salina Nauplii and Poecilia Reticulata Adults

Petroleum ether (pet. ether), chloroform (CHCl3) and methanol (CH3OH) extracts of Petunia integrifolia (Hook.) Schinz & Thell. Whole plant was subjected to assess cytotoxicity against the brine shrimp Artemia salina L. nauplii, larvicidal activity against the mosquito Culex quinquefasciatus Say larvae and piscicidal activity against the Poecilia reticulata Peters adults. The P. integrifolia extract in pet. ether showed the highest cytotoxic activity against A. salina nauplii by giving the minimum LC50 value of 49.25 parts per million (ppm) at 48 h, while in case of CHCl3 and CH3OH extracts, the values were 63.37 and 342.55 ppm, respectively. Against C. quinquefasciatus larvae the three extracts gave LC50 values of 4.92, 5.51 and171.26 ppm, respectively all after 48 h of exposure. Against P. reticulata the LC50 values were 21.70 ppm for pet. ether extract at 48 h, and 31.91 ppm for CHCl3 extract after 42 h of exposure but CH3OH extract offered no mortality. According to the intensity of activity, therefore, the extracts could be arranged in the descending order of: pet. ether > CHCl3 > CH3OH extracts; and the sensitivity of the test organisms to the extract of P. integrifolia could be arranged in the following descending order: C. quinquefasciatus larvae (in Pet. ether; LC50 = 4.92 ppm) > P. reticulata (in pet ether; LC50 = 21.70 ppm) >A. salina nauplii (in pet. ether; LC50 = 49.25 ppm) J. Bio-Sci. 31(1): 99-108, 2023

Genetic diversity and phylogenetic analyses of Asian lineage Zika virus whole genome sequences derived from Culex quinquefasciatus mosquitoes and urine of patients during the 2020 epidemic in Thailand

Zika virus (ZIKV), a mosquito-borne flavivirus, has been continually emerging and re-emerging since 2010, with sporadic cases reported annually in Thailand, peaking at over 1000 confirmed positive cases in 2016. Leveraging high-throughput sequencing technologies, specifically whole genome sequencing (WGS), has facilitated rapid pathogen genome sequencing. In this study, we used multiplex amplicon sequencing on the Illumina Miseq instrument to describe ZIKV WGS. Six ZIKV WGS were derived from three samples of field-caught Culex quinquefasciatus mosquitoes (two males and one female) and three urine samples collected from patients in three different provinces of Thailand. Additionally, successful isolation of a ZIKV isolate occurred from a female Cx. quinquefasciatus. The WGS analysis revealed a correlation between the 2020 outbreak and the acquisition of five amino acid changes in the Asian lineage ZIKV strains from Thailand (2006), Cambodia (2010 and 2019), and the Philippines (2012). These changes, including C-T106A, prM-V1A, E-V473M, NS1-A188V, and NS5-M872V, were identified in all seven WGS, previously linked to significantly higher mortality rates. Furthermore, phylogenetic analysis indicated that the seven ZIKV sequences belonged to the Asian lineage. Notably, the genomic region of the E gene showed the highest nucleotide diversity (0.7–1.3%). This data holds significance in informing the development of molecular tools that enhance our understanding of virus patterns and evolution. Moreover, it may identify targets for improved methods to prevent and control future ZIKV outbreaks.

Exploring the bioactive potential leaves of Sphaeranthus indicus: Targeting immature stages of Aedes aegypti and Culex quinquefasciatus mosquito vectors through bioassay-guided screening and fraction isolation

Mosquito control is vital for combating mosquito-borne diseases, but concerns exist regarding the use of synthetic insecticides. This study aimed to explore eco-friendly alternatives derived from natural sources. The larvicidal, pupicidal, and ovicidal activities of various fractions obtained from the hexane leaf extract of Sphaeranthus indicus were investigated against two important mosquito vectors, Aedes aegypti and Culex quinquefasciatus. S. indicus leaves were extracted with hexane and column chromatography was performed with hexane, ethyl acetate, methanol, and their mixtures as eluents. Among the ten fractions (F1-F10) evaluated, fraction ‘F-4’ exhibited significant activity against third instar larvae, pupae, and eggs of both mosquito species, closely followed by ‘F-5’ . At a concentration of 10 ppm, ‘F-4’ achieved 100% mortality in larvae and displayed LC50 values of 5.08 ppm and 5.03 ppm for Ae. aegypti and Cx. quinquefasciatus larvae, respectively. The LC50 values for pupae were 6.12 ppm and 5.83 ppm for Ae. aegypti and Cx. quinquefasciatus, respectively. Regarding ovicidal activity, ‘F-4’ demonstrated percentages ranging from 63.2% to 64.8% against Ae. aegypti and Cx. quinquefasciatus eggs, respectively. These findings underscore the potent larvicidal, pupicidal, and ovicidal effects of fraction ‘F-4’ from S. indicus against the targeted mosquito species. Further research is warranted to identify the active compounds responsible for these effects and explore practical applications for sustainable mosquito control strategies.

Elucidating the role of dsRNA sensing and Toll6 in antiviral responses of Culex quinquefasciatus cells.

The first step of any immune response is the recognition of foreign molecular structures inside the host organism. An important molecule that is generally foreign to eukaryotic cells is long double-stranded RNA (dsRNA), which can be generated during virus replication. The mechanisms of sensing viral dsRNA are well-studied in mammalian systems but are only poorly understood in insects, including disease vectors such as Culex quinquefasciatus mosquitoes. These mosquitoes are vectors for important arboviruses, such as West Nile virus, and Culex species mosquitoes are distributed across the globe in many temperate and tropical regions. The major antiviral response triggered by dsRNA in mosquitoes is RNA interference - a sequence-specific response which targets complementary viral RNA for degradation. However, here, we aimed to identify whether sequence-independent dsRNA sensing, mimicked by poly(I:C), can elicit an antiviral response. We observed a significant reduction in replication of La Crosse virus (LACV) in Cx. quinquefasciatus mosquito cells following poly(I:C) priming. We identified a number of antimicrobial peptides and Toll receptors that were upregulated at the transcript level by poly(I:C) stimulation. Notably, Toll6 was upregulated and we determined that a knockdown of Toll6 expression resulted also in increased LACV replication. Future efforts require genetic tools to validate whether the observed Toll6 antiviral activity is indeed linked to dsRNA sensing. However, large-scale functional genomic and proteomic approaches are also required to determine which downstream responses are part of the poly(I:C) elicited antiviral response.

Evaluation of field resistance in field-collected mosquito Culex quinquefasciatus Say through quantification of ULV permethrin/PBO formulation in field bioassays.

Pyrethroids are among the most applied adulticides worldwide to control mosquito vectors for prevention of arboviral diseases transmission. However, pesticide resistance development in a mosquito population could lead to decreased control efficacy. While most studies investigate the resistant genotype (i.e. kdr, CYP450, etc.) as explanatory variables, few field efficacy studies have measured pesticide quantities deposited at different distances from the sprayer in association with observed mosquito mortality. The current study determined field delivered amounts of an applied ULV permethrin/PBO formulation (31% permethrin + 66% piperonyl butoxide) by GC/MS and estimated practical resistance ratios using caged mosquito females. For field samples, the extraction method recovered 78 ± 3.92-108 ± 8.97% of the permethrin/PBO formulation when utilizing the peaks of PBO from GC/MS to estimate the concentrations of adulticide deposited near the mosquito cages. The field bioassay showed that the spatial distribution of permethrin/PBO formulation was heterogeneous among three pseudo-replicates within the same distance. Within the quantifiable permethrin/PBO range of 15.7-51.4 ng/cm2 , field-collected mosquito mortalities started at 64% and linearly increased reaching 100% only in two areas, while all Sebring susceptible mosquitoes died. The field LC95 resistance ratio (RR) of F0 Cx. quinquefasciatus ranged from 2.65-3.51, falling within the 95% CI of RR95 estimated by laboratory vial assays. Tests with and without PBO indicated P450's enzymes contributed to field resistance. Results showed the suitability of the collection and quantification method to estimate the field resistance ratio at the applied pesticide rate. Pesticide quantification would also allow the association of the known frequencies of resistance mechanisms (e.g. kdr, CYP450) with field mortalities to estimate the resistance level conferred by such mechanisms. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Toxic and synergetic effect of plant essential oils along with nano-emulsion for control of three mosquito species

Nano-emulsion containing Eucalyptus grandis (W. Hill ex Maiden) and Corymbia citrodora (Hook.) K.D.Hill & L.A.S.Johnson (common synonym Eucalyptus citriodora Hook.) leaf essential oils individual, combination was evaluated its larvicidal activity against Anopheles stephensi, Aedesaegypti and Culex quinquefasciatus mosquito larvae at 24 h post treatment. Nano-emulsion had low polydispersivity index and mean droplet size was below 66 nm. After 24 h E. grandis essential oil exhibited significant larvicidal activity with LC50 and LC90 values in A. aegypti (23.667; 438.797 ppm in A. aegypti A. stephensi (10.984; 277.535 ppm) and C. quinquefasciatus (4.676; 106.415 ppm). Similarly in C. citrodora showed low LC50 and LC90 values in A. aegypti (18.430; 1048.599 ppm), A. stephensi (6.573; 176.737 ppm) and C. quinquefasciatus (7.917; 287.720 ppm) respectively. The combination of E. grandis and C. citrodora essential oils showed higher mortality rates than individual essential oils, with low LC50 and LC90 values in A. aegypti (3.625; 58.962 ppm), A. stephensi (3.245; 34.473 ppm) and C. quinquefasciatus (3.002; 30.153 ppm). Nano-emulsion exhibited toxic effects similar to combination group with LC50 and LC90 values in A. aegypti (5.344; 113.362 ppm), A. stephensi (3.183; 83.112 ppm) and C. quinquefasciatus (2.832; 166.665 ppm). Chemical constituents of both essential oil were analyzed using GC-MS and FT-IR, result showed E. grandis contain different compounds namely; alpha-pinene, (31.74%); o-cymene, (17.84%); Eucalyptol (16.24%); Bicyclo[2.2.1]heptane (3.2%) and C. citrodora showed Citronellol (49.31%); dl-Isopulegol (25.39%) as a major compound while their synergistic action may be due to type’s composition of EOs might produce the synergist effect along with nanoemulsions. Our findings suggest that combination of essential oil E. grandis and C. citrodora can be used for mosquito control as green approach.

Isolation, Genomic, and Proteomic Characterization of a Novel Neotropical Strain of Bacillus thuringiensis with Mosquitocidal Activities

The combination of genomic and proteomic analyses is a useful tool for the study of novel Bacillus thuringiensis (Bt) strains, as these approaches allow the accurate identification of pesticidal proteins and virulence factors produced. Here, we isolated and evaluated the potential of a novel Neotropical Bt strain (TOD651) for controlling larvae of Aedes aegypti and Culex quinquefasciatus mosquitoes. Aiming for the full comprehension of the TOD651 larvicidal potential, we further evaluated the whole TOD651 genome and conducted the proteomic analysis of the TOD651 spore–crystal mixtures. Our results showed that Bt TOD651 similarly killed both A. aegypti (0.011 µg/mL) and C. quinquefasciatus (0.023 µg/mL) larvae, exhibiting similar potency to the commercial Bt strain. The genome sequence revealed that Bt TOD651 harbors cry11Aa3, cry10Aa4, cry4Aa4, cry4Ba5, cyt1Aa5, cyt1Ca1, cyt2Ba13, mpp60Aa3, and mpp60Ba3. The proteomic analysis revealed no expression of Mpp60Aa3, while all the other pesticidal proteins were expressed (Cry4Ba5 was more abundant than Cyt1Aa5). The expression of the Mppe showed the major proportions between proteases. The virulent factor neutral protease B and spore coat proteins were also expressed. The expression of relevant pesticidal proteins (e.g., Cry, Cyt, Mpp, and other pathogenic factors), whose actions can occur in a synergic relation, indicates that the biocontrol using Bt TOD651 may contribute to delaying the selection of resistant individuals.

Investigating insecticide susceptibility status of adult mosquitoes against some class of insecticides in Osogbo metropolis, Osun State, Nigeria.

The study evaluates the resistance and susceptibility of adult female Anopheles gambiae s. l., Aedes aegypti and Culex quinquefasciatus mosquitoes sourced within Osogbo metropolis, Osun State, Nigeria to four groups of insecticides [Permethrin, Deltamethrin, Pirimiphos-methyl and DDT (Dichlorodiphenyltrichloroethane)] and the distribution of their larval habitat within the metropolis. Mosquito larvae of the three genera were collected during the wet season and reared to adult stage in the laboratory. Emerged adult female mosquitoes were exposed to insecticide impregnated papers of the four insecticide groups for 60mins using WHO kits to determine the knock down rate (kdr). Thereafter, they were transferred into holding tubes and left for 24hrs to assess their resistance and susceptibility according to the WHO protocol. Four types of larval habitats were identified (tires, ground pools, gutters and plastic containers). Anopheles gambiae s. l. showed the highest resistance to Permethrin (49%) (p = 0.04, p<0.05) while the highest susceptibility was recorded with Pirimiphos-methyl (69%) with the lowest against Permethrin (16%) (P = 0.002; p<0.05). The highest resistance of A. aegypti was against OC-Control (45%) (p = 0.031; p<0.05). Permethrin had the highest susceptibility (60%) against A. aegypti while OC-control had the lowest (11%) (p = 0.005; p< 0.05). Culex quinquefasciatus had a lower resistance to OC-control (38%) as compared with Aedes aegypti (45%). However, it was least susceptible to Pirimiphos-methyl (52%) and DDT (17%) respectively (p = 0.013; p<0.05). The susceptibility of A. gambiae s. l. and C. quinquefasciatus to Pirimiphos-methyl and A. aegypti to Permethrin is an indication of the possibility of success if employed for vector control of A. gambiae s. l., C. quinquefasciatus and A. aegypti respectively. This could be through their inclusion as active ingredients in insecticide treated nets (ITNs) and indoor residual spray (IRS) with a view to abating malaria and other life-threatening mosquito-borne diseases constituting global public health scourge.

FIELD EVALUATION OF AUTOCIDAL GRAVID OVITRAP AND SIRENIX TRAP AGAINST CONTAINER INHABITING MOSQUITOES IN SAINT AUGUSTINE, NORTHEASTERN FLORIDA

Mosquito control programs are utilizing cost-effective long-term autocidal traps targeting the gravid population of container-inhabiting and other mosquito species, with the aim of reducing vector populations and disease transmission risk. In this field study we directly compared the efficacy of two autocidal trap types-the Autocidal Gravid Ovitrap (AGO) and SIRENIX mosquito trap in reducing mosquito abundances in St. Augustine, Florida to a control only site that had no autocidal traps deployed. Pre-treatment (wk1-4) and post-treatment (wk 5-14) adult mosquitoes were captured in all three sites using BG traps baited with BG lure and dry ice. Pre- and post-treatment trap counts of Aedes aegypti, Aedes albopictus, Culex quinquefasciatus, and total mosquitoes (three species together) were compared to determine significant changes in abundance. Percent reduction in abundance of each species/group at the two trap sites were calculated to evaluate the trap efficacy at controlling Aedes and Culex container mosquitoes. Aedes albopictus populations were significantly reduced (86.6%) at the SIRENIX site compared to the populations at the AGO site (67.7% reduction). Ae. aegypti populations were reduced by 72.4% at the SIRENIX site compared to 25% at the AGO site. Culex quinquefasciatus population reduction at the SIRENIX site was 59.6% compared to 11.8% at the AGO site. The total mosquito group had only 45.1% and 10.3% reduction at the SIRENIX and AGO sites, respectively. Furthers studies conducted across the entire mosquito season would be required for full understanding of the effectiveness of these traps.