Population genetic structure of Aedes aegypti subspecies in selected geographical locations in Sudan.

Although knowledge of the composition and genetic diversity of disease vectors is important for their management, this is limiting in many instances. In this study, the population structure and phylogenetic relationship of the two Aedes aegypti subspecies namely Aedes aegypti aegypti (Aaa) and Aedes aegypti formosus (Aaf) in eight geographical areas in Sudan were analyzed using seven microsatellite markers. Hardy-Weinberg Equilibrium (HWE) for the two subspecies revealed that Aaa deviated from HWE among the seven microsatellite loci, while Aaf exhibited departure in five loci and no departure in two loci (A10 and M201). The Factorial Correspondence Analysis (FCA) plots revealed that the Aaa populations from Port Sudan, Tokar, and Kassala clustered together (which is consistent with the unrooted phylogenetic tree), Aaf from Fasher and Nyala populations clustered together, and Gezira, Kadugli, and Junaynah populations also clustered together. The Bayesian cluster analysis structured the populations into two groups suggesting two genetically distinct groups (subspecies). Isolation by distance test revealed a moderate to strong significant correlation between geographical distance and genetic variations (p = 0.003, r = 0.391). The migration network created using divMigrate demonstrated that migration and gene exchange between subspecies populations appear to occur based on their geographical proximity. The genetic structure of the Ae. aegypti subspecies population and the gene flow among them, which may be interpreted as the mosquito vector's capacity for dispersal, were revealed in this study. These findings will help in the improvement of dengue epidemiology research including information on the identity of the target vector/subspecies and the arboviruses vector surveillance program.

Genetic Polymorphism and Phylogenetics of Aedes aegypti from Sudan Based on ND4 Mitochondrial Gene Variations.

This study investigated the genetic differences between Aedes aegypti subspecies (Aedes aegypti aegypti (Aaa) and Aedes aegypti formosus (Aaf)) from Sudan using the NADH dehydrogenase subunit 4 (ND4) mitochondrial gene marker. Nineteen distinct haplotypes of the ND4 were identified in female Aedes aegypti mosquitoes from the study sites. The phylogenetic relationship of the 19 ND4 haplotypes was demonstrated in a median-joining haplotype network tree with Aaa and Aaf populations found to share three haplotypes. The genetic variance (Pairwise FST values) was estimated and found to range from 0.000 to 0.811. Isolation by distance test revealed that geographical distance was correlated to genetic variation (coefficient value (r) = 0.43). The Polar maximum likelihood tree showed the phylogenetic relationship of 91 female Aaa and Aaf from the study sites, with most of the Aaf haplotypes clustered in one group while most of the Aaa haplotypes gathered in another group, but there was an admixture of the subspecies in both clusters, especially the Aaa cluster. The Spatial Analysis of Molecular Variance (SAMOVA) test revealed that the eight populations clustered into two phylogeographic groups/clusters of the two subspecies populations. The 2 Aedes aegypti subspecies seemed not to be totally separated geographically with gene flow among the populations.

Population genetics of Anopheles arabiensis, the primary malaria vector in the Republic of Sudan.

BACKGROUND: Anopheles arabiensis is a member of Anopheles gambiae complex and the main malaria vector in Sudan. There is insufficient population genetics data available on An. arabiensis for an understanding of vector population structure and genetics, which are important for the malaria vector control programmes in this country. The objective of this investigation is to study the population structure, gene flow and isolation by distance among An. arabiensis populations for developing control strategies. METHODS: Mosquitoes were collected from six sites located in three different states in Sudan, Khartoum, Kassala and Sennar, using pyrethrum spray catch of indoor resting mosquitoes. Anopheline mosquitoes were identified morphologically and based on species specific nucleotide sequences in the ribosomal DNA intergenic spacers (IGS). Seven published An. gambiae microsatellite loci primers were used to amplify the DNA of An. arabiensis samples. RESULTS: PCR confirmed that An. arabiensis was the main malaria vector found in the six localities. Of the seven microsatellite loci utilized, six were found to be highly polymorphic across populations, with high allelic richness and heterozygosity with the remaining one being monomorphic. Deviation from Hardy-Weinberg expectations were found in 21 out of 42 tests in the six populations due to heterozygote deficiency. Bayesian clustering analysis revealed two gene pools, grouping samples into two population clusters; one includes four and the other includes two populations. The clusters were not grouped according to the three states but were instead an admixture. The genetic distances between pairs of populations ranged from 0.06 to 0.24. Significant FST was observed between all pairwise analyses of An. arabiensis populations. The Kassala state population indicated high genetic differentiation (FST ranged from 0.17 to 0.24) from other populations, including one which is also located in the same state. High gene flow (Nm = 1.6-8.2) was detected among populations within respective clusters but limited between clusters particularly with respect to Kassala state. There was evidence of a bottleneck event in one of the populations (Al Haj Yousif site). No isolation by distance pattern was detected among populations. CONCLUSIONS: This study revealed low levels of population differentiation with high gene flow among the An. arabiensis populations investigated in Sudan, with the exception of Kassala state.

Distribution and Genetic Diversity of Aedes aegypti Subspecies across the Sahelian Belt in Sudan.

Aedes aegypti is the most important arboviral disease vector worldwide. In Africa, it exists as two morphologically distinct forms, often referred to as subspecies, Aaa and Aaf. There is a dearth of information on the distribution and genetic diversity of these two forms in Sudan and other African Sahelian region countries. This study aimed to explore the distribution and genetic diversity of Aedes aegypti subspecies using morphology and Cytochrome oxidase-1 mitochondrial marker in a large Sahelian zone in Sudan. An extensive cross-sectional survey of Aedes aegypti in Sudan was performed. Samples collected from eight locations were morphologically identified, subjected to DNA extraction, amplification, sequencing, and analyses. We classified four populations as Aaa and the other four as Aaf. Out of 140 sequence samples, forty-six distinct haplotypes were characterized. The haplotype and nucleotide diversity of the collected samples were 0.377-0.947 and 0.002-0.01, respectively. Isolation by distance was significantly evident (r = 0.586, p = 0.005). The SAMOVA test indicated that all Aaf populations are structured in one group, while the Aaa clustered into two groups. AMOVA showed 53.53% genetic differences within populations and 39.22% among groups. Phylogenetic relationships indicated two clusters in which the two subspecies were structured. Thus, the haplotype network consisted of three clusters.

Larval Age and Nutrition Affect the Susceptibility of Culex quinquefasciatus (Diptera: Culicidae) to Temephos.

Larval age and nutrition significantly affected the insect's physiology. These influences are important when rearing a population of vectors that is used to monitor the resistance level, in which standardized conditions are crucial for a more harmonized result. Little information has been reported on the effects of larval age and nutrition on the susceptibility of insects to insecticides, and therefore, we studied the effects on the susceptibility of Culex quinquefasciatus Say's (Diptera: Culicidae) larvae to temephos by comparing the median lethal concentration (LC50) after 24 hr between the second and fourth instar larvae and between the larvae that fed on protein-based and carbohydrate-based larval diets. The susceptibility of the larvae was significantly affected by the larval diets, as the larvae that fed on protein-based beef food and milk food demonstrated significantly higher LC50 value compared with the larvae that fed on carbohydrate-based food: lab food and yeast food. The larval diet interacted significantly with the larval age: while the second instar larvae were susceptible to temephos when supplied with carbohydrate-based food, the second and fourth instar larvae had no significant effect when supplied with protein-based diets, implying that a protein-rich environment may cause the mosquito to be less susceptible to temephos. This study suggested the importance of standardizing nutrition when rearing a vector population in order to obtain more harmonized dosage-response results in an insecticide resistance monitoring program. Future research could focus on the biochemical mechanism between the nutrition and the enzymatic activities of the vector.

Interaction Between Metarhizium anisopliae (Met.) and the Insecticides Used for Controlling House Fly (Diptera: Muscidae) in Poultry Farm of Malaysia.

The potential of integrating the mycoinsecticide, Metarhizium anisopliae (Met.), into house fly control programs is tremendous. However, the interaction between the fungus and insecticide, when applied at poultry farms, remains poorly understood. This study investigated the interaction between M. anisopliae and two selected insecticides, cyromazine and ChCy (a mixture of chlorpyrifos and cypemethrin), with three objectives: to assess the compatibility of M. anisopliae and the insecticides by measuring fungal vegetative growth and conidia production in the presence of insecticides; to evaluate the effect of M. anisopliae on these insecticides by analyzing insecticidal residue using ultra performance liquid chromatography; and to study the synergistic effects of M. anisopliae and the insecticides by applying sublethal concentrations of insecticides with M. anisopliae to house fly larvae. Metarhizium anisopliae was more tolerant to ChCy than to cyromazine, as M. anisopliae showed significantly more growth when grown with this insecticide. The M. anisopliae + ChCy combination resulted in significantly less chlorpyrifos residues compared to the ChCy plate, and 62-72% house fly larva mortality occurred when M. anisopliae and sublethal concentrations of ChCy were combined, implicating synergistic effects of the fungus with low concentrations of ChCy. Integrating M. anisopliae with compatible chemical at right concentration is crucial for poultry farm house fly control programs.

Comparison of Intrinsic Rate of Different House Fly Densities in a Simulated Condition: A Prediction for House Fly Population and Control Threshold.

Determining the control threshold for a pest is common prior to initiating a pest control program; however, previous studies related to the house fly control threshold for a poultry farm are insufficient for determining such a threshold. This study aimed to predict the population changes of house fly population by comparing the intrinsic rate of increase (rm) for different house fly densities in a simulated system. This study first defined the knee points of a known population growth curve as a control threshold by comparing the rm of five densities of house flies in a simulated condition. Later, to understand the interactions between the larval and adult populations, the correlation between larval and adult capacity rate (rc) was studied. The rm values of 300- and 500-fly densities were significantly higher compared with the rm values at densities of 50 and 100 flies. This result indicated their representative indices as candidates for a control threshold. The rc of larval and adult populations were negatively correlated with densities of fewer than 300 flies; this implicated adult populations with fewer than 300 flies as declining while the larval population was growing; therefore, control approaches should focus on the immature stages. The results in the present study suggest a control threshold for house fly populations. Future works should focus on calibrating the threshold indices in field conditions.

Molecular cloning, sequence analysis and developmental stage expression of a putative septin gene fragment from Aedes albopictus (Diptera: Culicidae).

Septins belong to GTPases that are involved in vital cellular activities, including cytokinesis. Although present in many organisms, they are yet to be isolated from Aedes albopictus. This study reports for the first time on a serendipitous isolation of a partial septin sequence from Ae. albopictus and its developmental expression profile. The Ae. albopictus partial septin sequence contains 591 nucleotides encoding 197 amino acids. It shares homology with several insect septin genes and has a close phylogenetic relationship with Aedes aegypti and Culex quinquefasciatus septins. The Ae. albopictus septin fragment was differentially expressed in the mosquito's developmental stages, with an increased expression in the adults.

Comparative Effectiveness of Insecticides for Use Against the House Fly (Diptera: Muscidae): Determination of Resistance Levels on a Malaysian Poultry Farm.

In this study, the toxicology of two commercial larvicides--cyromazine (Neporex 50SP) and ChCy (combination of chlorpyrifos and cypermethrin, Naga 505)--and five commercial adulticides--thiamethoxam (Agita 10WG), cyfluthrin (Responsar WP), lambda-cyhalothrin (Icon 2.8EC), fipronil (Regent 50SC), and imidacloprid (Toxilat 10WP)--was examined against the WHO/VCRU (World Health Organization/ Vector Control Research Unit) susceptible strain and the AYTW (Ayer Tawar) field strain of house fly, Musca domestica L. These pesticides were administered topically, in the diet, or as a dry residue treatment on plywood. Probit analysis using at least five concentrations and the concentration that was lethal to 50% (LC(50)) of the organisms was applied to compare the toxicology and resistance levels of the AYTW population to different insecticides. In the larvicide laboratory study, ChCy was more effective than cyromazine, with a significantly lower LC(50) value when administered topically or in the diet, although the AYTW population was susceptible to both larvicides with a resistance ratio (RR) <10. For the adulticide laboratory study, cyfluthrin and fipronil exhibited the lowest LC50 values of the adulticides, indicating that they are both effective at controlling adult flies, although lambda-cyhalothrin showed moderate resistance (RR = 11.60 by topical application; 12.41 by plywood treatment). Further investigation of ChCy, cyromazine, cyfluthrin, and fipronil under field conditions confirmed that ChCy and cyromazine strikingly reduced larval density, and surprisingly, ChCy also exhibited adulticidal activity, which significantly reduced adult fly numbers compared with the control group. Cyfluthrin and fipronil were also confirmed to be effective, with a significant reduction in adult fly numbers compared with the control group.

Paederus Outbreaks in Human Settings: A Review of Current Knowledge.

Although rove beetles (Paederus spp.) play a beneficial role as biological control agents to manage crop pests in agro-ecosystems, their high prevalence in human settings has elevated them to pest status in urban areas. Rove beetles neither bite nor sting, but accidental crushing on human skin causes them to release the toxin paederin, which causes dermatitis linearis. This review integrates currently available knowledge about the issues pertaining to Paederus infestation. For instance, the results of life history studies conducted under different food and temperature regimes are summarized, as they indicate how large a population can be in a habitat to cause massive and widespread infestation and illustrate the physiological traits required to maintain the population at the maximum level even under adverse conditions. In contrast to what is generally reported, we speculate that climatic factors do not necessarily result in Paederus dispersal in temperate regions; instead, habitat disturbance and site unsuitability may be the main factors that lead to massive dispersal to human settings. Factors such as whether dispersers are adaptable to xeric conditions in human settings, the probability that dispersed Paederus mate with the opposite sex, and whether dispersers have adequate nutrient intake to reproduce are considered to evaluate their potential to reproduce in human settings. Finally, the effectiveness of current commercial insecticides, challenges faced in managing infestations, and sustainable management practices are discussed to provide information for long-term control programs.

Contrasting patterns of insecticide resistance and knockdown resistance (kdr) in the dengue vectors Aedes aegypti and Aedes albopictus from Malaysia.

BACKGROUND: Knowledge on the extent, distribution and mechanisms of insecticide resistance is essential for successful insecticide-based dengue control interventions. Here, we report an extensive resistance profiling of the dengue vectors Aedes aegypti and Aedes albopictus across Malaysia and establish the contribution of knockdown resistance mechanism revealing significant contrast between both species. METHODS: Aedes mosquitoes were collected from four states in Malaysia in 2010 using ovitraps and tested against six major insecticides using WHO bioassays. Knockdown resistance (kdr) was investigated in both species. RESULTS: A moderate resistance to temephos was detected from samples collected in 2010 in Penang, Kuala Lumpur, Johor Bharu and Kota Bharu (1.5 < RR < 3.3). A widespread and multiple resistances was observed in Ae. aegypti particularly against pyrethroids, DDT and bendiocarb. Mosquitoes from Kuala Lumpur consistently had the highest resistance levels and was the only population showing a moderate resistance to malathion (91% mortality). The resistance profile of Ae. albopictus contrasted to Ae. aegypti with full susceptibility to pyrethroids except in Kuala Lumpur where moderate resistance is observed. PBO synergist assays suggest metabolic resistance mechanisms play a major role in resistance in both species. Two kdr mutations, F1534C and V1016G, were detected in Ae. aegypti across Malaysia but neither of these mutations were found in Ae. albopictus. Additionally, signatures of selection were detected on the Voltage-gated sodium channel gene in Ae. aegypti but not in Ae. albopictus. The presence of the 1534C allele was significantly associated with pyrethroid resistance and an additive effect to pyrethroid resistance was observed in individuals containing both kdr alleles. CONCLUSIONS: Findings from this study will help to design and implement successful insecticide-based interventions against Ae. aegypti and Ae. albopictus to improve dengue control across Malaysia.

Investigation of mosquito oviposition pheromone as lethal lure for the control of Aedes aegypti (L.) (Diptera: Culicidae).

BACKGROUND: The trend in chemical insecticide development has focused on improving the efficacy against mosquitoes while reducing the environmental impact. Lethal lures apply an "attract-and-kill" strategy that draws the insect to the killing agent rather than bringing the killing agent to the insect. METHODS: In this study, the mosquito oviposition pheromone was extracted from the eggs of Aedes aegypti (L.) and further investigated with a combination of pheromone and granular temephos as a lethal lure. RESULTS: The compound caproic acid attracted significantly more egg-laying mosquitos at 1 ppm (660.83 ± 91.61) than the control (343.83 ± 56.24), which consisted of solvent only (Oviposition Activity Index: 0.316). Further investigation of the combination of caproic acid with granular temephos as a lethal lure attracted significantly more gravid female Ae. aegypti to oviposit their eggs than the temephos treated water and control. CONCLUSIONS: This indicated the ability of caproic acid in acting as an attractant and counters the repellency effect of temephos. Additionally, the presence of temephos in the lethal lure also restricted the hatching of the eggs and killed any larvae that hatched.

Insecticidal activity and expression of cytochrome P450 family 4 genes in Aedes albopictus after exposure to pyrethroid mosquito coils.

Mosquito coils are insecticides commonly used for protection against mosquitoes due to their toxic effects on mosquito populations. These effects on mosquitoes could induce the expression of metabolic enzymes in exposed populations as a counteractive measure. Cytochrome P450 family 4 (CYP4) are metabolic enzymes associated with a wide range of biological activities including insecticide resistance. In this study, the efficacies of three commercial mosquito coils with different pyrethroid active ingredients were assessed and their potential to induce the expression of CYP4 genes in Aedes albopictus analyzed by real-time quantitative PCR. Coils containing 0.3 % D-allethrin and 0.005 % metofluthrin exacted profound toxic effects on Ae. albopictus, inducing high mortalities (≥90 %) compared to the 0.2 % D-allethrin reference coil. CYP4H42 and CYP4H43 expressions were significantly higher in 0.3 % D-allethrin treated mosquitoes compared to the other treated populations. Short-term (KT50) exposure to mosquito coils induced significantly higher expression of both genes in 0.005 % metofluthrin exposed mosquitoes. These results suggest the evaluated products provided better protection than the reference coil; however, they also induced the expression of metabolic genes which could impact negatively on personal protection against mosquito.

Effect of diet quality on survival and reproduction of adult Paederus fuscipes (Coleoptera: Staphylinidae).

The survival and reproduction of the beetle Paederus fuscipes Curtis on diets that differed in macronutrient composition were investigated in the laboratory. The sex organs of females fed each test diet were dissected to evaluate the ovarian development over time. Adults fed on a carbohydrate-rich diet lived longer than adults fed the other diets, but this diet provided insufficient nutrients for reproduction. Females fed on a protein-rich diet had high fecundity; however, the mean longevity of reproductive adult P. fuscipes significantly was shortened by 20-30 d compared with longevity of 60-70 d for adults fed a carbohydrate-rich diet. In contrast, adults that were provided lipid-rich diet had low survival and fecundity. Overall, the development ofprevitellogenic follicles was significantly affected by diet regime and days since starting a particular diet. The follicle size of females given protein-rich diet increased 0.5-fold 2 wk after beginning the diet. In contrast, the development of the follicles was slow in females given the carbohydrate-rich diet and the follicles degenerated in females given the lipid-rich diet. In terms of ovarian maturation, females fed on a protein-rich diet contained mostly vitellogenic and chorionated follicles. In contrast, the ovarioles of females on the carbohydrate-rich diet were largely occupied with previtellogenic and vitellogenic follicles, whereas for a lipid-rich diet, the follicles remained at the previtellogenic stage throughout the experiment.

Dispersal pattern of Paederus fuscipes (Coleoptera: Staphylinidae: Paederinae) in relation to environmental factors and the annual rice crop cycle.

Paederus fuscipes Curtis, a dermatitis linearis causing agent, has received increasing attention from the public, as it poses a serious health threat after mass dispersal into human-dominated areas. Preventive measures against this insect have so far been unsuccessful partly because of limited knowledge about its dispersal pattern. In this study, the dispersal activity of P. fuscipes was studied at infestation-prone residential buildings in Mainland Penang, Malaysia. The dispersal activity of P. fuscipes showed two peaks, that is, from February to April and August to October. Overall, there was no statistical significant correlation between dispersal and climatic parameters, that is, temperature, relative humidity, total rainfall, at all sampling localities. However, dispersal was primarily caused by human activities in rice fields, which accounted for >60% of the variability in dispersal. Particularly, rice harvesting, including straw burning, and cultivation were the major factors triggering P. fuscipes dispersal. These activities presumably disrupted the habitat and normal activities of P. fuscipes and rendered the rice fields unfavorable refuges. In addition, the beetles might also face food shortages after the disturbance of their prey base in the crop fields. The current study provides a predictive tool of P. fuscipes flight periods to ensure insecticide residual spraying is timed in the infestation-prone residential areas before the onset of infestation.

Influence of temperature on survival and water relations of Paederus fuscipes (Coleoptera: Staphylinidae).

The effects of four temperatures (15, 23.5, 28, and 35 degrees C) on the biological characteristics of the rove beetle Paederus fuscipes Curtis were studied, and its cuticular permeability also was measured. Specimens successfully developed to adulthood at each temperature tested, but development time of each preadult stage significantly decreased with increasing temperature. Both egg and L1 stages required at least 80 degree days above a threshold of approximately 10 degrees C to develop to the subsequent stage. The lengthy development time and high survival rate of preadults at 15 degrees C suggests that P. fuscipes can survive in a harsh environment during cold weather by hibernating, and this ability could allow preadults to succeed ecologically in temperate countries. However, adult longevity was short, and no fecundity was recorded at 15 degrees C. At 28 degrees C, P. fuscipes exhibited a high survival rate of adults, which had a longer life span and high fecundity; thus, the population had the highest intrinsic rate of increase (0.0788 +/- 0.0051 d(-1)) and the shortest mean generation time (48.57 +/- 1.43 d) at 28 degrees C. At this temperature, the population might reach a size that could facilitate invasion into residential areas. However, in the absence of a hygric environment, P. fuscipes was unable to survive despite favorable temperature. Unlike in adults and pupae, high cuticular permeability values were found in the larval stages. This indicates that larvae are highly susceptible to desiccation, and it explains why the distribution of P. fuscipes is restricted to moist habitats.

Bioefficacy of crude extract of Cyperus aromaticus (Family: Cyperaceae) cultured cells, against Aedes aegypti and Aedes albopictus mosquitoes.

OBJECTIVE: To evaluate the growth inhibition activity of the crude extract of Cyperus aromaticus (C. aromaticus) cultured cells against the 3rd instar larvae of Aedes aegypti (Linn.) and Aedes albopictus Skuse (Ae. albopictus) under laboratory conditions, and determine the sublethal effects (EI50) of the crude extract of C. aromaticus cultured cells on some biological and morphological parameters of both Aedes mosquito species during two generations as well. METHODS: The cell suspension cultures of C. aromaticus were activated from five callus lines (P4, Pa, Z1, Z6 and Ml) derived from the root explants of in vitro plantlets. The cultured cells were extracted in chloroform and used as plant material for the present study. For detection of juvenile hormone III, the crude extracts were analyzed by HPLC. Then the crude extracts of the three C. aromaticus cultured cell lines which contained varied amounts of juvenile hormone III [high level (P4 cell line), medium level (Z1 cell line) and low level (Ml cell line)] were tested against Aedes mosquito species. Laboratory evaluation was performed against late third instar larvae of the Vector Control Research Unit strains of Ae. aegypti and Ae. albopictus using the standard WHO method. The effects of EI50 of the C. aromaticus cultured P4 cells on fecundity, fertility, growth period, sex ratio, adult size and longevity of Aedes mosquitoes were assessed. RESULTS: Bioassay tests presented the remarkable growth inhibition activity of the crude extracts of C. aromaticus cultured cells against the two Aedes mosquitoes. Between the two mosquito species, Ae. albopictus was more susceptible to the crude extracts with lower EI50 values. EI50 of the crude extract of C. aromaticus cultured cells (P4) increased the sterility indices in the parental generation females in both Aedes mosquito species. A significant delay in the pupal formation and adult emergence were observed in the parental generation of the both mosquito species. The sex ratio of the adult population either parental or F1 generation of the Aedes mosquito species was not significantly affected by the EI50 dosage of the crude extract of C. aromaticus cultured P4 cells. A significant decrease in the wing length of the treated adult (female and male) of Aedes aegypti as well as the treated female of Ae. albopictus were observed. Longevity of the adult female of the parental generation of both Aedes mosquitoes as well as females of F1 generation of Ae. albopictus were significantly decreased. CONCLUSIONS: The present study revealed the potential of the crude extract of C. aromaticus cultured cells in controlling vector mosquito populations in the effort to reduce the transmission of vector borne diseases.

Larvicidal, oviposition, and ovicidal effects of Artemisia annua (Asterales: Asteraceae) against Aedes aegypti, Anopheles sinensis, and Culex quinquefasciatus (Diptera: Culicidae).

This study focuses on the larvicidal, oviposition, and ovicidal effects of a crude extract of Artemisia annua against Aedes aegypti, Anopheles sinensis, and Culex quinquefasciatus. Dried cells of Artemisia annua from cell suspension cultures were extracted using hexane. The extract showed moderate larvicidal effects against mosquitoes. At 24-h post treatment, the LC50 values for Anopheles sinensis, Aedes aegypti, and Culex quinquefasciatus were recorded as 244.55, 276.14, and 374.99 ppm, respectively. The percentage mortality of larvae was directly proportional to the tested concentration. Anopheles sinensis was found to be the most susceptible species, whereas Culex quinquefasciatus was the most tolerant to the Artemisia annua extract. The results indicated that the Artemisia annua extract showed concentration-dependent oviposition deterrent activity and had a strong deterrent effect. At 500 ppm, the percentage effective repellency was more than 85% compared with the control group for all the species, with oviposition activity index values of -0.94, -0.95, and -0.78 for Aedes aegypti, Anopheles sinensis, and Culex quinquefasciatus, respectively. In the ovicidal assay, the percentage hatchability of eggs after treatment with 500 ppm of Artemisia annua extract was significantly lower than the control, with values of 48.84 ± 4.08, 38.42 ± 3.67, and 79.35 ± 2.09% for Aedes aegypti, Anopheles sinensis, and Culex quinquefasciatus, respectively. Artemisia annua was found to be more effective against Aedes aegypti and Anopheles sinensis compared with Culex quinquefasciatus. This study indicated that crude extract of A. annua could be a potential alternative for use in vector management programs.

Contact toxicity and residual effects of selected insecticides against the adult Paederus fuscipes (Coleoptera: Staphylinidae).

The contact toxicity of four insecticide formulations (deltamethrin, fipronil, fenitrothion, and imidacloprid) applied on three different substrates (tile, plywood, and concrete) against the adult rove beetle, Paederus fuscipes Curtis, was evaluated. The relative order of speed of killing effects was as follows: deltamethrin > imidacloprid > fipronil > fenitrothion. Although deltamethrin showed the fastest action against P. fuscipes, the recovery rate of rove beetles at 48 h posttreatment was moderate (approximately 25%) on the tile surface to high (approximately 80%) on the plywood surface. Thus, it is likely that the insects did not pick up the lethal dose especially on porous surfaces. In contrast, fipronil demonstrated delayed toxicity that might promote maximal uptake by the insects. More than 80% mortality was registered for tile and plywood surfaces up to 4 wk after exposure. High mortality (almost 100%) was recorded for imidacloprid-exposed P. fuscipes at 48 h posttreatment, but only on the tile surface. Among the four insecticides tested, fenitrothion was the least effective against P. fuscipes because low percentage to no mortality was recorded in the fenitrothion treatment.

Bioefficacy of crude extract of Cyperus aromaticus (Family：Cyperaceae) cultured cells, against Aedes aegypti and Aedes albopictus mosquitoes

Objective:To evaluate the growth inhibition activity of the crude extract of Cyperus aromaticus (C. aromaticus) cultured cells against the 3rd instar larvae of Aedes aegypti (Linn.) and Aedes albopictus Skuse (Ae. albopictus) under laboratory conditions, and determine the sublethal effects (EI50) of the crude extract of C. aromaticus cultured cells on some biological and morphological parameters of both Aedes mosquito species during two generations as well. Methods:The cell suspension cultures of C. aromaticus were activated from five callus lines (P4, Pa, Z1, Z6 and Ml) derived from the root explants of in vitro plantlets. The cultured cells were extracted in chloroform and used as plant material for the present study. For detection of juvenile hormone III, the crude extracts were analyzed by HPLC. Then the crude extracts of the three C. aromaticus cultured cell lines which contained varied amounts of juvenile hormone III [high level (P4 cell line), medium level (Z1 cell line) and low level (Ml cell line)] were tested against Aedes mosquito species. Laboratory evaluation was performed against late third instar larvae of the Vector Control Research Unit strains of Ae. aegypti and Ae. albopictus using the standard WHO method. The effects of EI50 of the C. aromaticus cultured P4 cells on fecundity, fertility, growth period, sex ratio, adult size and longevity of Aedes mosquitoes were assessed. Results:Bioassay tests presented the remarkable growth inhibition activity of the crude extracts of C. aromaticus cultured cells against the two Aedes mosquitoes. Between the two mosquito species, Ae. albopictus was more susceptible to the crude extracts with lower EI50 values. EI50 of the crude extract of C. aromaticus cultured cells (P4) increased the sterility indices in the parental generation females in both Aedes mosquito species. A significant delay in the pupal formation and adult emergence were observed in the parental generation of the both mosquito species. The sex ratio of the adult population either parental or F1 generation of the Aedes mosquito species was not significantly affected by the EI50 dosage of the crude extract of C. aromaticus cultured P4 cells. A significant decrease in the wing length of the treated adult (female and male) of Aedes aegypti as well as the treated female of Ae. albopictus were observed. Longevity of the adult female of the parental generation of both Aedes mosquitoes as well as females of F1 generation of Ae. albopictus were significantly decreased. Conclusions: The present study revealed the potential of the crude extract of C. aromaticus cultured cells in controlling vector mosquito populations in the effort to reduce the transmission of vector borne diseases.