Larvicidal activity of Pseudocalymma alliaceum and Allium sativum against Culex quinquefasciatus (Say)

The larvicidal activity of the plant extracts Pseudocalymma alliaceum and Allium sativum were determined against Culex quinquefasciatus. The hexane extract of P. alliaceum and the petroleum ether extract of A. sativum exhibited larvicidal efficacy against Cx. quinquefasciatus larvae. Extracts of P. alliaceum resulted in concentrations that produced 50% mortality LC₅₀ and LC₉₀ values of 2.49 and 15.06 ppm, respectively, after 24 h and 1.16 and 8.45 ppm after 48 h. Extracts of A. sativum resulted in LC₅₀ and LC₉₀ values of 8.38 and 29.15 ppm after 24 h and 7.28 and 44.19 ppm after 48 h of exposure, respectively. The results indicate that the plant extract component(s) present in the hexane extract of P. alliaceum leaves demonstrated greater potential as an efficient larvicide than A. sativum against Cx. quinquefasciatus.     

光活化多炔类化合物对蚊幼虫的毒力

采用自制的光活化实验装置,测定了11个合成的多炔类化合物对致倦库蚊Culex quinquefasciatus 4龄幼虫的光活化毒力,发现部分化合物在近紫外光照射条件下,能明显地提高光活化毒杀效应,测得化合物5(1-苯基-4-(3,4-亚甲基二氧)苯基-丁二炔)光照与未光照处理LC₅₀分别是0.35 μg/mL和8.89 μg/mL。实验中发现蚊虫先接触药后,再进行光照处理,才能较好地发挥毒效,而且毒杀效应与光照时间呈正相关。模拟田间试验表明,太阳光能显著提高化合物5毒杀蚊幼虫的药效。利用抗氧化剂进行猝灭作用试验,间接地证明化合物5的光活毒杀机理是与过氧化作用有关。分析结构与活性关系,发现二苯基-丁二炔衍生物比二烷基取代丁二炔活性高,苯基上不同取代基也影响光活毒杀效果,它们的活性顺序是：亚甲基二氧基>甲氧基>邻硝基>间硝基>甲基酯。     

Larvicidal efficacy of some Cucurbitacious plant leaf extracts against Culex quinquefasciatus (Say)

Larvicidal efficacies of extracts of five species of Cucurbitacious plants, Momordica charantia, Trichosanthes anguina, Luffa acutangula, Benincasa cerifera and Citrullus vulgaris were tested against the late third larval age group of Culex quinquefasciatus. The larval mortality was observed after 24 h exposure. The LC50 values of M. charantia, T. anguina, L. acutangula, B. cerifera and C. vulgaris were 465.85, 567.81, 839.81, 1189.30 and 1636.04 ppm respectively.     

Comparative responses of ovipositing Anopheles gambiae and Culex quinquefasciatus females to the presence of Culex egg rafts and larvae

Field observations have demonstrated that gravid Anopheles gambiae Giles s.s. (Diptera: Culicidae) are selective in their choice of oviposition sites. For example, immature stages of An. gambiae s.s. are rarely found in water that contains Culex quinquefasciatus Say immatures. The possibility that this may, in part at least, reflect a response by ovipositing An. gambiae s.s. females to volatile signals associated with Culex juveniles was evaluated by testing the response of An. gambiae s.s. females to varying densities of Cx. quinquefasciatus egg rafts and/or larvae in oviposition choice assays. For comparison, the oviposition choices of Cx. quinquefasciatus to conspecific egg rafts and/or larvae were similarly assayed. At a low density of Cx. quinquefasciatus egg rafts (1-15 egg rafts/100 mL water), An. gambiae s.s. females laid more eggs in the treatment water than in the control, with a maximum of twice as many in the treatment water at 5 egg rafts/100 mL water. At higher egg raft densities and in all treatments that included Cx. quinquefasciatus larvae, oviposition decreased significantly in the treatment dishes in a density-dependent manner. As previous studies have indicated, ovipositing Cx. quinquefasciatus females were attracted to and laid egg rafts in dishes containing conspecific egg rafts and, interestingly, also in dishes containing larvae.     

Differential effects of Bacillus sphaericus strain 2362 on Culex quinquefasciatus and its competitor Culex cinereus in West Africa

The larval susceptibility to Bacillus sphaericus strain 2362 of the non-man-biting mosquito Culex cinereus and the urban filariasis vector Culex quinquefasciatus, two competitor mosquitoes in polluted habitats, was compared. In the laboratory, both species ingested a similar amount of B. sphaericus spores when fed c. 2 x 10(5) spores per ml for 30 min. However, in the same experiment, third-instar larvae of Cx quinquefasciatus were reduced by 98% at 24 h exposure while Cx cinereus larvae were only reduced by 6% at 72 h. In the field, preimaginal populations of Cx cinereus ingested, within a week, more than 99% of the applied spores, but showed no significant decrease through 14 days in cesspools treated at 10 g/m2 of a flowable concentrate of B. sphaericus 2362, containing 2 x 10(10) spores/g. It is proposed that specific biological control of Cx quinquefasciatus could result from appropriate treatment of breeding-sites with larvicidal B. sphaericus and competitive displacement by Cx cinereus or other mosquitoes with larvae that are more tolerant of B. sphaericus.     

Larvicidal potency of selected xerophytic plant extracts on Culex pipiens (Diptera: Culicidae)

Chemical insecticides released into the environment may have adverse biological effects. Therefore, there is a need for ecofriendly insecticides for mosquito control. Xerophytic plant extracts that may provide more ecofriendly active component were evaluated against Culex pipiens 4th instars. Plant extracts prepared using different solvents with a Soxhlet apparatus and different concentrations were tested against Culex pipiens larvae. The effects were observed at 24 h and 72 h intervals and LD₅₀ and LD₉₀ values determined. Chloroform (CHCl₃) and ethyl acetate (EtOAc) extracts of Althaea ludwigii were the most effective against Cx. pipiens 4^th instars, but were highly dependent on extract concentrations and exposure time. Results suggest that A. ludwigii extracts contain bioactive compounds, such as phenols and saponins, that may provide effective Cx. pipiens larval control. However, the extract was found to be toxic to zebrafish larvae, and may be toxic to other aquatic fauna. Further studies to determine the active components and toxicity to other fauna are needed.     

Stempellia milleri sp. n. (Microsporida: Nosematidae) in the Mosquito Culex pipiens quinquefasciatus Say*

Stempellia milleri sp. n. was found in blood cells and the adipose tissue of field-collected larvae of Culex pipiens quinquefasciatus Say. Its development is different from other Stempellia species described in that it is dimorphic, producing two types of spores in adipose cells. Both were studied with the electron microscope. One spore is thin-walled and uninucleate; the other is thick-walled and binucleate. The latter spores are produced from sporonts that are somewhat similar to those of Nosema. Sporoblasts and spores of S. milleri were compared to those of Stempellia magna Kudo, and Stempellia lunata Hazard and Savage in both light and electron microscope preparations. S. milleri was transmitted experimentally to Culex pipiens pipiens, C. p. quinquefasciatus, C. salinarius, C. tarsalis, and C. territans. Transmission of S. magna to its host and other mosquito species was not possible. Although, generally low numbers of test larvae became infected with the pathogen, heavy infections were seen occasionally in individual specimens of its natural host, C. p. quinquefasciatus. Species of Aedes, Anopheles, Culiseta, Psorophora, and Uranotaenia exposed to spores of S. milleri were not susceptible to the disease.     

Influence of light and habitat on predation of Culex quinquefasciatus (Diptera: Culicidae) larvae by the waterbugs (Hemiptera: Heteroptera)

The influence of light and habitat structure on the predation of Culex quinquefasciatus larvae by the common heteropteran water bug, Diplonychus (=Sphaerodema) annulatus, D. rusticus and Anisops bouvieri was assessed in the laboratory. It was revealed that water bugs predate more in presence of light than in dark conditions. While A. bouvieri consumed more prey in structured conditions, D. annulatus and D. rusticus consumed more prey in open conditions. The selection of prey size as well as the respective numbers varied between predators and treatments significantly. Prey vulnerability (PV), an indicator of predatory efficiency, was highest for D. annulatus, moderate for D. rusticus and low for A. bouvieri. Prey consumption and PV values under different treatment conditions indicate that for belostomatid water bugs D. annulatus and D. rusticus, the order of prey consumption under different habitat and light/dark combinations is light open > dark open > light vegetated > dark vegetated. In the case of the backswimmer, A. bouvieri, the order of prey consumption appeared to be light structured > dark structured > light open > dark open. These findings were consistent with resource‐partitioning by water bug species sharing the same guild. If the observed results are extended to natural settings the efficiency of these predators in controlling mosquito populations will vary with the structural complexity of habitats and the intensity of light.     

Effects of blood meal source on the reproduction of Culex pipiens quinquefasciatus (Diptera: Culicidae)

Culex pipiens quinquefasciatus were fed blood meals from a live chicken (LC), chicken blood in Alsever's (AC) solution, defibrinated bovine blood (DB), or bovine blood in citrate (CB) and incubated at 28° C. The effects of different blood meal sources were evaluated with respect to rates of blood feeding and reproduction (i.e., fecundity and fertility) over two gonotrophic cycles. Mosquitoes that fed on the first blood meal were subjected to a second blood meal as follows (first blood meal / second blood meal): LC/LC, LC/DB, DB/DB, CB/CB, AC/AC. Fecundity and fertility of Cx. p. quinquefasciatus were significantly (P < 0.05) greater in mosquitoes fed LC blood; however, fecundity and fertility in different treatment groups varied by gonotrophic cycle. These results contribute to our understanding of the impact of blood meal source on feeding and reproduction in Cx. p. quinquefasciatus. The potential impacts of blood meal source on virus transmission experiments are discussed.     

Narrow-sense heritability of body size and its response to different developmental temperatures in Culex quinquefasciatus (Say 1923)

Body size is an important trait involved in overall fitness through its effects on mating success, fecundity, resource acquisition and mortality, and desiccation resistance. In this study, we raised inbred Culex quinquefasciatus mosquito cohorts at different developmental temperatures of 20°, 23°, and 27° C. As an indicator of the amount of genetic variation in body size, we estimated the narrow-sense heritability of body sizes defined as wing aspect ratios. Our results show that narrow-sense heritability of the body size increased as the developmental temperature increased. We also detected the presence of strong genotype-by-environment (G × E) interaction from low cross-environmental correlations. The body size of each temperature regime followed the general rule that higher temperatures produce smaller individuals. We suggest that the increase in genetic variation with increasing temperature might be due to an unleashing of the cryptic genetic variation of the putative genes affecting body size. We conclude that this increase in genetic variation tracking the environmental (developmental temperature) change could have considerable implications for the distribution and range expansion of Cx. quinquefasciatus, especially in warmer environments.     

Flight muscle-specific expression of act88F: GFP in transgenic Culex quinquefasciatus Say (Diptera: Culicidae)

A strategy to engineer a strain of Culex mosquitoes refractory to filarial transmission is described. A requirement for success of the strategy is identification of a flight muscle-specific promoter that functions in the mosquito. A GFP marker gene under the control of the promoter region of the Drosophila melanogaster act88F gene was inserted into the genome of Culex quinquefasciatus. Transformation was confirmed by Mendelian genetics. Hybridization of a genomic Southern blot to a radiolabeled probe verified that the entire donor plasmid integrated into the mosquito genome. GFP expression in the transgenic mosquitoes was restricted to the flight muscles.     

Distribution of intramembranous particles and filipin-sterol complexes in the spermatid and spermatozoon of Culex quinquefasciatus (Culicidae).

A freeze-fracture study was carried out on spermatid and spermatozoon of the mosquito Culex quinquefasciatus. In the spermatid plasma membrane few and randomly distributed intramembranous particles were observed. In the spermatozoon the density of intramembranous particles was higher on the P- than on the E-fracture face of the plasma membrane. Two populations of particles were observed. Large particles (about 15 nm in diameter) are regularly arranged in double rows as a zipper-line, longitudinally oriented in relation to the main cell axis. These strands of particles were observed in the posterior head region, mainly associated with the E-fracture face. Filipin was used to analyse the presence and distribution of cholesterol in thin sections and freeze-fracture replicas. Filipin-sterol complexes were not homogeneously distributed throughout the spermatozoon plasma membrane. They were more abundant on the P-fracture face of the membrane lining the nuclear region. The results obtained show that Culex spermatozoon differs from those of other species in that its plasma membrane exhibits only a membrane domain, the zipper-line, localized in the postacrosomal region.     

Larvicidal Activity of Essential Oils of Five Apiaceae Taxa and Some of Their Main Constituents Against Culex quinquefasciatus

Apiaceae are aromatic herbs producing essential oils which are used on an industrial scale for various purposes. Notably, Apiaceae essential oils may replace synthetic insecticides keeping most of their efficacy and avoiding environmental pollution and human poisoning. In the present work, we explored the insecticidal potential of the essential oils from five Apiaceae taxa, namely Sison amomum, Echinophora spinosa, Heracleum sphondylium subsp. sphondylium, Heracleum sphondylium subsp. ternatum, and Trachyspemum ammi, as well as their major constituents (sabinene, p‐cymene, terpinolene, myristicin, and thymol), against the filariasis vector Culex quinquefasciatus. For the purpose, the essential oils were obtained by hydrodistillation and their composition was achieved by gas chromatography/mass spectrometry (GC/MS). Their acute toxicity on third instar larvae of C. quinquefasciatus was determined. The two most active essential oils were those from T. ammi fruits and E. spinosa roots, showing LC₅₀ below 20 μl/l and LD₉₀ below 50 μl/l. These oils were dominated by the monoterpene phenol thymol and the phenylpropanoid myristicin, respectively, which showed the strongest larvicidal activity (LC₅₀ of 15.1 and 16.3 μl/l, respectively) among the pure compounds tested. These results showed that Apiaceae may be useful as source of larvicidal compounds to be used for the development of cheap, effective and eco‐friendly insecticidal formulations.     

Evaluation of Solanum xanthocarpum extract as a synergist for cypermethrin against larvae of the filarial vector Culex quinquefasciatus (Say)

Cypermethrin and crude extracts of Solanum xanthocarpum were both observed for their larvicidal activity against Culex quinquefasciatus. Petroleum ether extract with lethal concentration (LC)₅₀ and LC₉₀ of 41.28 and 111.16 p.p.m. after 24 h and LC₅₀ 38.48 and LC₉₀ 80.83 p.p.m. after 48 h, respectively, was found to be the most effective, followed by carbon tetrachloride and methanol extracts. LC₅₀ and LC₉₀ for cypermethrin were 0.0027 and 0.0097 p.p.m. after 24 h and 0.0013 and 0.0092 p.p.m. after 48 h of exposure, respectively. Combined formulations were evaluated for synergistic activity and a 1:1 ratio of cypermethrin and petroleum ether extract was observed to be more effective than 1:2 and 1:4 ratios. Combinations of S. xanthocarpum extracts and cypermethrin demonstrated higher larvicidal activity, indicating synergistic activity. These results demonstrate the need for further studies on the effectiveness and toxicity to humans and animals, particularly aquatic forms.     

Field cage insecticide resistance tests against Culex quinquefasciatus Say (Diptera: Culicidae) in Harris County,Texas, U.S.A.

The ground adulticiding program in densely populated Harris County has been employed on the basis of virus‐positive mosquitoes almost exclusively using the ‘rotation of chemicals best practices’ stratagem. To evaluate its effectiveness, 15 comparative field cage testing events were conducted from 2011–2015 using seven wild population samples from repeatedly collected locations of Culex quinquefasciatus Say and a laboratory susceptible Sebring strain colony reared to adults. A 3 × 3 plot design was employed for exposure to ultra‐low volume applications of malathion and synergized permethrin. No significant differences were found in mortality rate among testing dates, year, row placement, or relative humidity. Mortality was significantly different between adulticides (p<0.001) with mean mortality rates for malathion 96.42% (±7.95%) and permethrin 92.38% (±14.04). There was a significant temperature difference for permethrin (p<0.001) but none for malathion (p=0.644). Mosquito population mortality was statistically different by study operational area (p<0.011) and chemical (p<0.001). Susceptible colony adults used as positive controls downwind strongly aided determination of efficacy and resistance to each adulticide, providing evidence of individual application coverage, though comparative analysis was done with overall mortalities by normal methodology.     

Age modifies the effect of body size on fecundity in Culex quinquefasciatus Say (Diptera: Culicidae)

Fecundity of mosquitoes can vary with many factors and can have a strong effect on population growth. This study reports the effects of body size, blood meal size, and age on the reproductive output of nulliparous Culex quinquefasciatus, a vector of arboviruses and other pathogens. Mated adult female mosquitoes from a colony were reared under standard conditions and fed on chickens at different ages post‐eclosion. Blood meal size and wing length were recorded, as well as the number of eggs in the first‐cycle egg raft. Each of these factors had a significant influence on fecundity considered in a simple regression context. Multiple regression analysis revealed a significant interaction effect between age and body size on fecundity. Up to 13 days of age, fecundity was positively correlated with body size, but in mosquitoes older than 13 days, this relationship was not significantly different from zero. These results are discussed in terms of the known physiology of this and other species.     

Estimating reaction norms for predictive population parameters,age specific mortality,and mean longevity in temperature‐dependent cohorts of Culex quinquefasciatus Say (Diptera: Culicidae)

Culex quinquefasciatus plays a major role in the transmission of important parasites and viruses throughout the world. Because temperature is an important limiting factor on growth and longevity of all mosquito species, estimating the reaction norms provides very important basic information for understanding both plasticity and individual variations of the population. In the present study, Cx. quinquefasciatus were maintained at five different constant temperatures (15°, 20°, 23°, 27°, and 30°C) for two subsequent generations. Reproductive population parameters in blood‐fed mated females and longevities of virgin and blood‐fed mated adults reared at different temperatures were compared for the two generations. Longevity increased as temperature decreased within a range of 15° to 30°C for the unmated adults, and 15° to 27°C for the mated and blood‐fed adults. Generation times were as long as 124.07 and 106.76 days for two subsequent generations reared at 15°C, and the highest intrinsic rate of increase (r_m) values were estimated at 0.22 and 0.18, respectively, from the cohorts reared at 27°C. For survival rates, reproductive rates (R₀), and r_m values, 30°C was found to be a critical temperature for this species. These cohorts produced the smallest amount of eggs (R₀= 5.06), r_m values decreasing across generations (from 0.11 to 0.06), and the survival rates from egg to adult were found to be insufficient (16.1 and 10.8%). Additionally, the rate of exponential increase with age and age specific mortalities (b) were calculated for the virgin cohorts. Age specific mortality rates increased as temperature decreased. The increase in mortality rates started to accelerate at 27°C and was more pronounced at 30°C, for both females and males. We estimated the coefficients of variation for the b values in which females have smaller coefficients than those of the males at all temperatures.     

Permethrin resistance variation and susceptible reference line isolation in a field population of the mosquito,Culex quinquefasciatus (Diptera: Culicidae)

This study examines the genetic variations and mechanisms involved in the development of permethrin resistance in individual mosquitoes from a field population of Culex quinquefasciatus, HAmCq^G0, and characterizes susceptible reference lines of mosquitoes with a similar genetic background to the field HAmCq^G0 strain. Six upregulated cytochrome P450 genes, CYP9M10, CYP9J34, CYP6P14, CYP9J40, CYP6AA7, and CYP4C52v1, previously identified as being upregulated in the larvae of resistant HAmCq^G8 mosquitoes were examined in the larvae of 3 strains (susceptible S‐Lab, parental HAmCq^G0 and permethrin‐selected highly resistant HAmCq^G8) and 8 HAmCq^G0 single‐egg raft colonies, covering a range of levels of susceptibility/resistance to permethrin and exhibiting different variations in the expression of A and/or T alleles at the L‐to‐F kdr locus of the sodium channel. The 2 lines with the lowest tolerance to permethrin and bearing solely the susceptible A allele at the L‐to‐F kdr locus of the sodium channels, from colonies Cx_SERC5 and Cx_SERC8, showed lower or similar levels of all 6 of the P450 genes tested compared with the S‐Lab strain, suggesting that these 2 lines could be used as the reference mosquitoes in future studies characterizing insecticide resistance in HAmCq mosquitoes. This study also provides a detailed investigation of the mechanisms involved in insecticide resistance in individuals within a population: individuals with elevated levels of resistance to permethrin all displayed one or more potential resistance mechanisms–either elevated levels of P450 gene expression, or L‐to‐F mutations in the sodium channel, or both.