Laboratory evaluation of lactic acid on attraction of Culex spp. (Diptera: Culicidae)

The role of lactic acid was evaluated for attraction of Culex nigripalpus, Culex quinquefasciatus, Culex tarsalis, and Aedes aegypti in the laboratory using a dual‐port olfactometer. When lactic acid was combined with chicken odor, attraction was increased for Cx. quinquefasciatus compared to chicken odor alone but not for Cx. nigripalpus, Cx. tarsalis, and Ae. aegypti. Lactic acid combined with hand odor did not change attraction of Cx. tarsalis and Ae. aegypti but decreased attraction of Cx. nigripalpus and Cx. quinquefasciatus. The addition of lactic acid to CO₂ increased attraction of Ae. aegypti and Cx. quinquefasciatus but reduced attraction of Cx. nigripalpus and Cx. tarsalis. Use of commercial lactic acid baits with CO₂ resulted in a similar trend except for Cx. nigripalpus which showed no difference. A blend of lactic acid, acetone, and dimethyl disulfide was attractive to Ae. aegypti (63.4%) but elicited low responses by all Culex spp. (1.3–26.8%). Addition of the blend to CO₂ increased attraction of Ae. aegypti and Cx. quinquefasciatus but reduced attraction of Cx. nigripalpus and Cx. tarsalis. The mixture of compounds plus CO₂ was as attractive as a hand for Cx. quinquefasciatus, Cx. tarsalis, and Ae. aegypti.     

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.     

Butterflies of the South Pacific

ABSTRACT

Larvae of the invasive mosquito Culex quinquefasciatus Say are morphologically similar to those of the native Culex pervigilans Bergroth, yet distinguishing these species can be hampered by morphological variations in Cx. quinquefasciatus. We present detail about the extent of these variations in an urban population of Cx. quinquefasciatus in Auckland. To aid in identification of this exotic species, we provide images of key diagnostic characters and some observed exceptions to these. Details regarding variation in diagnostic characters for < 3rd instar and 3rd/4th instar larvae are given. Of the nine characters used for identification, three were highly consistent (dorsal papillae, mantle plate, pecten teeth); each observed in > 90% of larvae, although these characters were not always visible. Other characters were less reliable, for instance, the expected position of seta 1a-S in relation to the pecten teeth was observed in < 10% of larvae. Further exploration of regional morphological variation in both Cx. quinquefasciatus and Cx. pervigilans is recommended, ideally with associated molecular characterisation.     

Toxicity, transformation and accumulation of inorganic arsenic species in a microalga Scenedesmus sp. isolated from soil

Arsenic speciation and cycling in the natural environment are highly impacted via biological processes. Since arsenic is ubiquitous in the environment, microorganisms have developed resistance mechanisms and detoxification pathways to overcome the arsenic toxicity. This study has evaluated the toxicity, transformation and accumulation of arsenic in a soil microalga Scenedesmus sp. The alga showed high tolerance to arsenite. The 72-h 50 % growth inhibitory concentrations (IC₅₀ values) of the alga exposed to arsenite and arsenate in low-phosphate growth medium were 196.5 and 20.6 mg? L^?1, respectively. When treated with up to 7.5 mg? L^?1 arsenite, Scenedesmus sp. oxidised all arsenite to arsenate in solution. However, only 50 % of the total arsenic remained in the solution while the rest was accumulated in the cells. Thus, this alga has accumulated arsenic as much as 606 and 761 μg? g^?1 dry weight when exposed to 750 μg? L^?1 arsenite and arsenate, respectively, for 8 days. To our knowledge, this is the first report of biotransformation of arsenic by a soil alga. The ability of this alga to oxidise arsenite and accumulate arsenic could be used in bioremediation of arsenic from contaminated water and soil.     

Survival of West Nile virus‐challenged Southern house mosquitoes,Culex pipiens quinquefasciatus,in relation to environmental temperatures

We investigated the effect of West Nile virus (WNV) infection on survival in two colonies of Culex pipiens quinquefasciatus Say (Diptera: Culicidae) originating from Vero Beach and Gainesville, FL. Mosquitoes were fed West Nile virus‐infected blood and checked daily for survival. Exposure to WNV decreased survival among Cx. p. quinquefasciatus from Gainesville relative to unexposed individuals at 31° C. In contrast, exposure to WNV enhanced survival among Cx. p. quinquefasciatus from Vero Beach relative to unexposed individuals at 27° C. These results may suggest that exposure to WNV and associated infection could increase or decrease components of fitness, dependent on environmental temperature and intraspecific variation in Cx. p. quinquefasciatus. The relationship between lifespan (time of death in days) and WNV titer differed in the colonies at 31° C and 27° C, suggesting that intraspecific species variation in response to temperature impacts interactions with WNV. While further work is needed to determine if similar effects occur under field conditions, this suggests intraspecific variation in vector competence for WNV and adult survival of Cx. p. quinquefasciatus, both aspects of vectorial capacity.     

Larvicidal potentials of leaf and bark extracts of Eucalyptus camaldulensis (Schlect) and Eucalyptus citriodora (Hook) on Culex quinquefasciatus (Say) larvae

The larvicidal activity of leaf and bark extracts of Eucalyptus camaldulensis and Eucalyptus citriodora on juveniles of Culex quinquefasciatus was determined. Larvae were successfully bred in the laboratory to obtain a “pure” colony. One hundred and sixty‐two batches of 25 larvae each were treated with 3.90, 15.63, 62.50, 250 and 1000 mg/L of plant extract in three replicates, for 24 h to record mortality and for up to 96 h to record feeding and fecundity. There was an increase in percentage mortality with increase in concentration of the petroleum ether plant extracts. There was a statistically significant difference (P < 0.05) between extracts used in the bioassay and mortality of larvae, but there was no significant difference (P > 0.05) between the two plant species. Statistical analysis of average mortality figures using Probit analysis for the leaf extracts of E. citriodora and E. camaldulensis, respectively, had LC₅₀ of 245.47 (26.89 ± 10.13 [SE]) and 316.23 (22.89 ± 8.95) for the crude extracts; 97.72 (44.95 ± 12.89) and 223.87 (29.17 ± 9.56) for the petroleum ether extracts; and 162.18 (30.7 ± 11.16) and 257.04 (26.47 ± 9.28) for the methanol extracts. The bark extracts of E. citriodora and E. camaldulensis, respectively, had LC₅₀ of 251.19 (27.78 ± 8.39) and 398.11 (20.7 ± 7.17) for methanol; and 302.0 (21.56 ± 8.11) and 630.96 (13.56 ± 6.4) for hexane. Both plant species showed anti‐feeding properties against larvae of Culex quinquefasciatus.     

Mosquitoes and West Nile virus along a river corridor from prairie to montane habitats in eastern Colorado

We conducted studies on mosquitoes and West Nile virus (WNV) along a riparian corridor following the South Platte River and Big Thompson River in northeastern Colorado and extending from an elevation of 1,215 m in the prairie landscape of the eastern Colorado plains to 1,840 m in low montane areas at the eastern edge of the Rocky Mountains in the central part of the state. Mosquito collection during June‐September 2007 in 20 sites along this riparian corridor yielded a total of 199,833 identifiable mosquitoes of 17 species. The most commonly collected mosquitoes were, in descending order: Aedes vexans, Culex tarsalis, Ae. dorsalis, Ae. trivittatus, Ae. melanimon, Cx. pipiens, and Culiseta inornata. Species richness was higher in the plains than in foothills‐montane areas, and abundances of several individual species, including the WNV vectors Cx. tarsalis and Cx. pipiens and the nuisance‐biter and potential secondary WNV vector Ae. vexans, decreased dramatically from the plains (1,215‐1,487 m) to foothills‐montane areas (1,524‐1,840 m). Ae. vexans and Cx. tarsalis had a striking pattern of uniformly high abundances between 1,200‐1,450 m followed by a gradual decrease in abundance above 1,450 m to reach very low numbers above 1,550 m. Culex species were commonly infected with WNV in the plains portion of the riparian corridor in 2007, with 14 of 16 sites yielding WNV‐infected Cx. tarsalis and infection rates for Cx. tarsalis females exceeding 2.0 per 1,000 individuals in ten of the sites. The Vector Index for abundance of WNV‐infected Cx. tarsalis females during June‐September exceeded 0.5 in six plains sites along the South Platte River but was uniformly low (0–0.1) in plains, foothills and montane sites above 1,500 m along the Big Thompson River. A population genetic analysis of Cx. tarsalis revealed that all collections from the ≈190 km riparian transect in northeastern Colorado were genetically uniform but that these collections were genetically distinct from collections from Delta County on the western slope of the Continental Divide. This suggests that major waterways in the Great Plains serve as important dispersal corridors for Cx. tarsalis but that the Continental Divide is a formidable barrier to this WNV vector.     

Microbial oxidation of arsenite in a subarctic environment: diversity of arsenite oxidase genes and identification of a psychrotolerant arsenite oxidiser

Background  

Arsenic is toxic to most living cells. The two soluble inorganic forms of arsenic are arsenite (+3) and arsenate (+5), with arsenite the more toxic. Prokaryotic metabolism of arsenic has been reported in both thermal and moderate environments and has been shown to be involved in the redox cycling of arsenic. No arsenic metabolism (either dissimilatory arsenate reduction or arsenite oxidation) has ever been reported in cold environments (i.e. < 10°C).     

Resistance potential of Culex quinquefasciatus against the insect growth regulators methoprene and diflubenzuron

Bioassay procedures for the insect growth regulators diflubenzuron (a chitin synthetase inhibitor) and methoprene (a juvenile hormone mimic) were evaluated on the mosquito Culex quinquefasciatus Say to determine the most sensitive larval stages and the effects of varying exposure periods. Sensitivity to diflubenzuron decreased gradually during instars I and II; instars III and IV were relatively insusceptible. Sensitivity to methoprene increased with larval age. Standard test procedures were devised for diflubenzuron involving continuous exposure from the early third instar and for methoprene as 6 h exposure during instar IV. Selection for ten generations of strains from Dar-es-Salaam which were multiresistant to insecticides increased the LC₅₀ to diflubenzuron by 2.8-fold and increased the LC₅₀ to methoprene by 3.9-fold. When compared to a standard susceptible strain from Colombo the resistance ratios of Dar-es-Salaam strains rose from 2.4 x to 6.6 x for diflubenzuron and from 6.1 x to 21.3 x for methoprene, with no significant effects detected on the pattern of cross-resistance. These results are discussed in relation to standardization of IGR testing and the need for integrated control programmes against Cx quinquefasciatus.

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Résistance potentielle de Culex quinquefasciatus Say au méthoprène et au diflubenzuron, régulateurs de croissance des insectes

Résumé La détermination du stade larvaire le plus sensible et les effects de différentes durées d'exposition aux régulateurs de croissance des insectes-diflubenzuron (inhibiteur de la chitine synthétase) et méthoprène (mimétique de l'hormone juvénile)—ont été recherchés sur C. quinquefasciatus. La sensibilité au diflubenzuron diminue graduellement pendant les deux premiers stades, les stades III et IV étant relativement insensibles. La sensibilité au méthoprène augmente avec l'âge des larves. Les essais comportaient une exposition continue au diflubenzuron depuis le début du stade III et une exposition de 6 heures au méthoprène pendant le stade IV. La sélection pendant 10 générations de lignées de Dares-Salam multirésistantes aux insecticides a entraîné une augmentation due 2,8 fois de la DL 50 au diflubenzuron et de 3,9 fois au méthoprène. Comparés aux souches standards sensibles de Colombo, les taux de résistance des lignées de Dar-es-Salam se sont élevés de 2,4 à 6,6 pour le diflubenzuron et de 6,1 à 21,3 pour le méthoprène, sans effet visible significatif sur la résistance croisée. Ces résultats sont discutés en relation avec la standardisation du test IGR et les besoins de programmes de lutte intégrée contre C. quinquefasciatus.

     

Influence of resource levels,organic compounds and laboratory colonization on interspecific competition between the Asian tiger mosquito Aedes albopictus (Stegomyia albopicta) and the southern house mosquito Culex quinquefasciatus

The mosquitoes Aedes albopictus (Stegomyia albopicta) (Skuse) and Culex quinquefasciatus (Say) (Diptera: Culicidae) are common inhabitants of tyres and other artificial containers, which constitute important peridomestic mosquito breeding habitats. We tested the hypotheses that interspecific resource competition between the larvae of these species is asymmetrical, that the concentration of chemicals associated with decomposing detritus affects the competitive outcomes of these species, and that wild and colonized strains of Cx. quinquefasciatus are affected differently by competition with Ae. albopictus. We conducted two laboratory competition experiments wherein we measured survivorship and estimated population growth (λ′) in both species under multiple mixed‐species densities. Under varying resource levels, competition was asymmetrical: Ae. albopictus caused competitive reductions or exclusions of Cx. quinquefasciatus under conditions of limited resources. In a second experiment, which used both wild and colonized strains of Cx. quinquefasciatus, organic chemical compounds associated with decomposing detritus did not affect the competitive outcome. The colonized strain of Cx. quinquefasciatus had greater survivorship and adult mass, and faster development times than the wild strain, but both strains were similarly affected by competition with Ae. albopictus. Competition between these species may have important consequences for vector population dynamics, especially in areas in which tyres and artificial containers constitute the majority of mosquito breeding habitats.     

Permethrin and malathion LD90 values for Culex quinquefasciatus vary with topical application site

Prior research in multiple insect species has demonstrated that insecticide‐induced mortality varies according to the body region exposed on the insect. This variation has been demonstrated in Culex quinquefasciatus Say (Diptera: Culicidae), but has not been quantified using dose–response curves. Applications of technical permethrin or malathion to one of three body regions on Cx. quinquefasciatus resulted in dose–response curves that were not equivalent to one another. The generated LD₉₀ values and curves for each body region were compared with previously reported LD values for analogous sites in several mosquito species, specifically the mesothorax. Based on the present results, the permethrin and malathion LD₅₀ and LD₉₀ concentrations required for droplets impinging on the abdomen and mesothorax of Cx. quinquefasciatus when applied through ground‐based spray systems utilized by mosquito control programmes were calculated.     

Insecticide susceptibility and resistance of Culex tritaeniorhynchus (Diptera: Culicidae) larvae collected from Gwangju,Republic of Korea

The susceptibility of Culex tritaeniorhynchus collected from Gwangju, Jeollabuk Province, Republic of Korea (ROK) to insecticides was evaluated under laboratory conditions using ten insecticides (7 pyrethroids and 3 organophosphates) that are currently applied by local public health centers in the ROK. Based on the values of median lethal concentration (LC₅₀), Cx. tritaeniorhynchus larvae were most susceptible to chlorpyrifos (0.006 ppm), fenitrothion (0.022 ppm), fenthion (0.035 ppm) and bifenthrin (0.038 ppm), and were least susceptible to esbiol (1.722 ppm). In comparative resistance tests, the resistance ratios (RRs) of seven insecticides were compared among each other using two strains of Cx. tritaeniorhynchus that were collected from the same locality during 1992 and 2010. Culex tritaeniorhynchus demonstrated significantly increased RRs to pyrethroids over time, while demonstrating decreased RRs among the organophosphates. Among the pyrethroids, permethrin had the highest RR values of 182.1‐ and 833.3‐fold differences, followed by etofenprox with RRs of 138.4‐ and 224.1‐fold differences in values of LC₅₀ and concentration that produced 90% mortality (LC₉₀), respectively. Culex tritaeniorhynchus strains demonstrated the least amount of change in susceptibility to the organophosphates, chlorpyrifos, fenitrothion and fenthion with 0.020‐, 0.019‐ and 0.001‐fold differences in resistance ratios (RRLC₅₀), respectively.     

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

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

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.     

Oviposition preference and offspring performance in container breeding mosquitoes: evaluating the effects of organic compounds and laboratory colonisation

1. The preference–performance hypothesis (PPH) predicts that organisms lacking parental care should oviposit in habitats that optimise offspring performance. Preference–performance relationships were investigated for the Asian tiger mosquito (Aedes albopictus Skuse) and the southern house mosquito (Culex quinquefasciatus Say) (Diptera: Culicidae), two medically important container‐breeding species, in response to an organic chemical blend mimicking decaying plant matter. Additionally, the effects of long‐term laboratory colonisation of Cx. quinquefasciatus using wild and laboratory strains were evaluated. 2. Oviposition bioassays were conducted by releasing gravid mosquitoes into field enclosures with automobile tires containing low and high concentrations of the chemical blend, and water controls. The offspring were then reared in water collected from the tires in which they were deposited. 3. Aedes albopictus and wild Cx. quinquefasciatus laid more eggs in the chemical blend than water controls but did not differentiate between the low and high concentrations. Conversely, laboratory Cx. quinquefasciatus only preferred the high concentration to the low concentration. No statistical associations between oviposition preference and larval survival were found, as the chemical blend did not affect survivorship of either species. 4. The oviposition preference for the chemical blend over water controls suggests that both species oviposit in the best available resource environment, but further studies are needed before conclusions regarding preference–performance relationships can be drawn. 5. It was found that long‐term laboratory colonisation affects the oviposition behaviour in Cx. quinquefasciatus, suggesting that behavioural studies on laboratory strains are not always applicable to wild populations.     

Mitochondrial DNA analyses reveal low genetic diversity in Culex quinquefasciatus from residential areas in Malaysia

The present study explored the intraspecific genetic diversity, dispersal patterns and phylogeographic relationships of Culex quinquefasciatus Say (Diptera: Culicidae) in Malaysia using reference data available in GenBank in order to reveal this species' phylogenetic relationships. A statistical parsimony network of 70 taxa aligned as 624 characters of the cytochrome c oxidase subunit I (COI) gene and 685 characters of the cytochrome c oxidase subunit II (COII) gene revealed three haplotypes (A1–A3) and four haplotypes (B1–B4), respectively. The concatenated sequences of both COI and COII genes with a total of 1309 characters revealed seven haplotypes (AB1–AB7). Analysis using tcs indicated that haplotype AB1 was the common ancestor and the most widespread haplotype in Malaysia. The genetic distance based on concatenated sequences of both COI and COII genes ranged from 0.00076 to 0.00229. Sequence alignment of Cx. quinquefasciatus from Malaysia and other countries revealed four haplotypes (AA1–AA4) by the COI gene and nine haplotypes (BB1–BB9) by the COII gene. Phylogenetic analyses demonstrated that Malaysian Cx. quinquefasciatus share the same genetic lineage as East African and Asian Cx. quinquefasciatus. This study has inferred the genetic lineages, dispersal patterns and hypothetical ancestral genotypes of Cx. quinquefasciatus.     

Differential protein expression in the midgut of Culex quinquefasciatus mosquitoes induced by the insecticide temephos

Mosquitoes are vectors for pathogens of malaria, lymphatic filariasis, dengue, chikungunya, yellow fever and Japanese encephalitis. Culex quinquefasciatus Say, 1823 (Diptera: Culicidae) is a known vector of lymphatic filariasis. Its control in Brazil has been managed using the organophosphate temephos. Studies examining the proteins of Cx. quinquefasciatus that are differentially expressed in response to temephos further understanding of the modes of action of the insecticide and may potentially identify resistance factors in the mosquito. In the present study, a comparative proteomic analysis, using 2‐dimensional electrophoresis coupled with matrix‐assisted laser desorption/ionization (MALDI) time of flight (TOF)/TOF mass spectrometry, and bioinformatics analyses were performed to identify midgut proteins in Cx. quinquefasciatus larvae that were differentially expressed in response to exposure to temephos relative to those in untreated controls. A total of 91 protein spots were differentially expressed; 40 were upregulated and 51 were downregulated by temephos. A total of 22 proteins, predominantly upregulated, were identified as known to play a role in the immune response, whereas the downregulated proteins were involved in energy and protein catabolism. This is the first proteome study of the midgut of Cx. quinquefasciatus and it provides insights into the molecular mechanisms of insecticide‐induced responses in the mosquito.     

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.