Pathogenicity of entomopathogenic fungus,Metarhizium anisopliae MET-GRA4 isolate on dengue vectors,Aedes albopictus and Aedes aegypti mosquito larvae (Diptera: Culicidae)

Considering the rapid transmission of the dengue virus, substantial efforts need to be conducted to ward-off the epidemics of dengue viruses. The control effort is depending on chemical insecticides and had aroused undesirable conflicts of insecticide resistance. Here, we study the entomopathogenic fungus, Metarhizium anisopliae as a promising new biological control agent for vector control. The pathogenicity effects of Metarhizium anisopliae against field and laboratory strains of Aedes albopictus and Aedes aegypti larvae were tested using the larvicidal bioassay technique. The results demonstrate that the treatments using M. anisopliae isolate MET-GRA4 were highly effective and able to kill 100% of both Ae. albopictus and Ae. aegypti mosquito larvae at a conidia concentration of 1 × 10?/ml within 7 days of the treatment period. The fungus displayed high larvicidal activity against laboratory and field strain of Ae. aegypti larvae with LC₅₀ values (9.6 × 10³/ml, 1.3 × 10³/ml) and LC₉₅ values (1.2 × 10?/ml, 5.5 × 10⁵/ml) respectively. For Ae. albopictus, LC₅₀ values for laboratory and field strains were (1.7 × 10⁴/ml, 2.7 × 10⁴/ml) and the LC₉₅ values were (2.1 × 10?/ml, 7.0 × 10⁵/ml) respectively. Interestingly, the susceptibility of field strain towards M. anisopliae was higher as compared to the laboratory strain Aedes larvae. In which, the causative agents of all the dead larvae were verified by the virulence of M. anisopliae and caused morphological deformities on larval body. The findings from this study identify this isolate could be an effective potential biocontrol agent for vector mosquitoes in Malaysia.     

Evaluation of the toxicity of different phytoextracts of Ocimum basilicum against Anopheles stephensi and Culex quinquefasciatus

The larvicidal effect of the crude carbon tetrachloride, methanol and petroleum ether leaf extracts of a widely grown medicinal plant, Ocimum basilicum, against Anopheles stephensi and Culex quinquefasciatus was evaluated. Petroleum ether extract was found to be the most effective against the larvae of both mosquitoes, with LC₅₀ values of 8.29, 4.57; 87.68, 47.25 ppm and LC₉₀ values of 10.06, 6.06; 129.32, 65.58 ppm against A. stephensi and C. quinquefasciatus being observed after 24 and 48 h of treatment, respectively. The efficacy of petroleum ether was followed by that of the carbon tetrachloride and methanol extracts, which had LC₅₀ values of 268.61, 143.85; 446.61, 384.84 ppm and LC₉₀ values of 641.23, 507.80; 923.60, 887.00 ppm against A. stephensi after 24 and 48 h, respectively, and LC₅₀ values of 24.14, 17.02; 63.48, 53.77 ppm and LC₉₀ values of 295.38, 204.23; 689.71, 388.87 ppm against C. quinquefasciatus after 24 and 48 h of treatment, respectively. These extracts are highly toxic against mosquito larvae from a range of species; therefore, they may be useful for the management of mosquito larvae to control vector borne diseases.     

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.     

Synthesis of some novel phosphorylated and thiophosphorylated benzimidazoles and benzothiazoles and their evaluation for larvicidal potential to Aedes albopictus and Culex quinquefasciatus

Series of benzimidazole and benzothiazole linked phosphoramidates and phosphoramidothioates (5a–j) and benzimidazole linked phenylphosphoramidates and phenylphosphoramidothioates (10a–e) were synthesized. The title compounds were preliminary screened for mosquito larvicidal properties against Aedes albopictus and Culex quinquefasciatus at different concentration from 40 to 5 mg/L. Among the screened compounds three compounds revealed potential larvicidal effects with 100% mortality in the order of 10e > 5j > 5e. Compound 10e was found to be the most toxic compound to Ae. albopictus and Cx. quinquefasciatus. The LC₅₀ of 10e against Ae. albopictus was found to be 6.42 and 5.25 mg/L at 24 and 48 h, respectively, whereas it was 7.01 and 3.88 mg/L, respectively in Cx. quinquefasciatus. Temephos was used as positive control.     

Purification,characterization and larvicidal activity of a potent bioactive compound asarone from leaves of Acorus calamus against the culician larval mosquitoes

Mosquitoes are potent vectors by serving as agents to life-threatening diseases in humans. Increasing resistance in mosquitoes against existing insecticides and repellents brings new challenges and an opportunity to explore sustainable compounds. We chose six medicinal plants to screen potential bioactive compounds that could act as an insecticide. Among these, crude hexane leaf extract of Acorus calamus showed higher mortality percentage against Aedes aegypti and Culex quinquefasciatus. The LC₅₀ and LC₉₀ values were 151.86 ppm and 536.36 ppm, respectively, for the third instar A. aegypti larvae, and 174.70 ppm and 696.73 ppm, respectively, for C. quinquefasciatus. The treated larvae of both species showed morphological and physiological variations when compared to control. The GC–MS profile of purified fractions showed a single peak. Further, FT-IR and NMR analyses confirmed the propensity of the purified compound as trans asarone (phenylpropanoid; C₁₂H₁₆O₃. LC₅₀ and LC₉₀ values of purified asasone-treated larvae were 2.35 ppm and 12.58 ppm, respectively, for A. aegypti and 2.15 ppm and 11.58 ppm, respectively, for C. quinquefasciatus. Treatment of different sub-lethal doses of asarone to mosquito larvae at various time intervals showed disruption of intestinal layers. By showing negligible toxicity to non-target organism, purified asarone has a great potential in vector management.     

An effective entomopathogenic fungus Metarhizium anisopliae for the simultaneous control of Aedes albopictus and Culex pipiens mosquito adults

The purpose of this study was to select the most effective Metarhizium anisopliae isolates for the simultaneous control of Aedes albopictus and Culex pipiens mosquito adults. The selection was performed using 65 isolates of M. anisopliae, an entomopathogenic fungus isolated from Korean soils. The selection process was carried out through evaluations of conidial thermotolerance, UV-B tolerance, pathogenicity and virulence against adults of two mosquito species and conidial production on grain medium. The thermotolerance of 65 M. anisopliae isolates ranged from 0% to 99%, and their UV-B tolerance ranged from 7.1% to 94.8%. Among them, 30 isolates with high thermotolerance were selected for evaluation of their pathogenicity and virulence against adults of the two mosquito species. Metarhizium anisopliae isolates were more susceptible to Cx. pipiens adults than Ae. albopictus adults because 11 isolates were pathogenic to Ae. albopictus and 21 isolates were pathogenic to Cx. pipiens. Virulence and conidial production were evaluated using 6 isolates with similar pathogenicity against the two species of mosquito. As a result, the accumulated mortality was 74–100% for Ae. albopictus and 78–100% for Cx. pipiens. LT₅₀ values ranged from 2.3 to 5.2 days for Ae. albopictus and from 3.2 − 4.7 days for Cx. pipiens. Comparison of conidial production using three kinds of grain media showed various results according to isolate and medium. Based on the above results, M. anisopliae CN6S1W1 was finally selected as the most effective fungal isolate for the simultaneous control of Ae. albopictus and Cx. pipiens mosquito adults.     

Formulation of the entomopathogenic fungus Beauveria bassiana JN5R1W1 for the control of mosquito adults and evaluation of its novel applicability

In this study, an effective control agent and control technology for adult mosquitoes using entomopathogenic fungi is proposed as a solution to the adverse effects of chemical insecticides used for mosquito control. A formulation study was conducted using Beauveria bassiana JN5R1W1 with high virulence against both Aedes albopictus and Culex pipiens adult mosquitoes. For the inverted emulsion formulation, three kinds of vegetable oils were used to evaluate the germination promotion, UV blocking effects, and adhesion rate of conidia. As a result, soybean oil was selected as the most effective vegetable oil, and the corresponding formulation was made. Direct contact treatment of the inverted emulsion by spraying resulted in a mortality rate of approximately 100 % for both species of adult mosquitoes. However, in reality, since it is difficult to directly apply fungal agent treatment against adult mosquitoes, we evaluated a control method by indirect contact with mosquitoes after applying the inverted emulsion on a net. As an indirect treatment method, the prepared inverted emulsion showed a mortality rate of approximately 93 % or 89 % against Ae. albopictus and Cx. pipiens adult mosquitoes, respectively. Therefore, these results suggested that the inverted emulsion using B. bassiana JN5R1W1 is an effective control agent for controlling adult mosquitoes.     

Unraveling microbe-mediated interactions between mosquito larvae in a laboratory microcosm

Interspecies interactions have important impacts on communities and when multiple trophic levels are involved, effects can be complex and indirect. For mosquitoes, interactions experienced as larvae affect adult attributes such as survivorship, reproductive output, and longevity, factors that can affect their ability to vector disease. We examined how larvae of two ecologically distinct mosquito species, Aedes japonicus japonicus and Culex quinquefasciatus, interact at different temperatures (17 and 27 °C) and at different relative densities. We also quantified abundances of bacteria and protozoan flagellates to uncover how changes in the microbial community affect the outcome of the two mosquitoes’ interaction. At 17 °C, survival and size of both mosquito species were not affected by the other’s presence. Cx. quinquefasciatus was strongly affected by intraspecific, but not interspecific, competition at both temperatures. At 27 °C, Ae. j. japonicus larvae experienced 100 % mortality in treatments by themselves and treatments where Cx. quinquefasciatus was abundant, surviving only in the presence of low densities of Cx. quinquefasciatus. Both the total bacteria count and counts of a protozoan flagellate identified as Spumella spp. decreased with increasing numbers of Cx. quinquefasciatus. We postulate that at 27 °C, the survival of Ae. j. japonicus depends on the interaction between Cx. quinquefasciatus and the microbial community. This study demonstrates that one mosquito species may alter the microbial community in ways that indirectly influence another mosquito species’ larval survival, and by extension adult abundance and potential disease transmission.     

Larvicidal and mosquito repellent action of peppermint (Mentha piperita) oil

Oil of Mentha piperita L. (Peppermint oil), a widely used essential oil, was evaluated for larvicidal activity against different mosquito species: Aedes aegypti, Anopheles stephensi and Culex quinquefasciatus by exposing IIIrd instar larvae of mosquitoes in enamel trays 6 × 4 inch² size filled to a depth of 3 inch with water. Of the three species tested Cx. quinquefasciatus was most susceptible followed by Ae. aegypti and An. stephensi. Application of oil at 3 ml/m² of water surface area resulted in 100% mortality within 24 h for Cx. quinquefasciatus, 90% for Ae. aegypti and 85% for An. stephensi. For Ae. aegypti 100% mortality was achieved at 3 ml/m² in 48 h or 4 ml/m² in 24 h. For An. stephensi 100% mortality was observed at 4 ml/m² in 72 h. The emergence at 3 ml/m² was also inhibited to a great extent and the few adults which emerged did not ovipost even after taking a blood meal. The oil showed strong repellent action against adult mosquitoes when applied on human skin. Percent protection obtained against An. annularis, An. culicifacies, and Cx. quinquefasciatus was 100%, 92.3% and 84.5%, respectively. The repellent action of Mentha oil was comparable to that of Mylol oil consisting of dibutyl and dimethyl phthalates.     

Ongoing challenges in the management of malaria

Background

Mosquitoes transmit serious human diseases, causing millions of deaths every year. Use of synthetic insecticides to control vector mosquitoes has caused physiological resistance and adverse environmental effects in addition to high operational cost. Insecticides of botanical origin have been reported as useful for control of mosquitoes. Azadirachta indica (Meliaceae) and its derived products have shown a variety of insecticidal properties. The present paper discusses the larvicidal activity of neem-based biopesticide for the control of mosquitoes.

Methods

Larvicidal efficacy of an emulsified concentrate of neem oil formulation (neem oil with polyoxyethylene ether, sorbitan dioleate and epichlorohydrin) developed by BMR & Company, Pune, India, was evaluated against late 3^rd and early 4^th instar larvae of different genera of mosquitoes. The larvae were exposed to different concentrations (0.5–5.0 ppm) of the formulation along with untreated control. Larvicidal activity of the formulation was also evaluated in field against Anopheles, Culex, and Aedes mosquitoes. The formulation was diluted with equal volumes of water and applied @ 140 mg a.i./m² to different mosquito breeding sites with the help of pre calibrated knapsack sprayer. Larval density was determined at pre and post application of the formulation using a standard dipper.

Results

Median lethal concentration (LC₅₀) of the formulation against Anopheles stephensi, Culex quinquefasciatus and Aedes aegypti was found to be 1.6, 1.8 and 1.7 ppm respectively. LC₅₀ values of the formulation stored at 26°C, 40°C and 45°C for 48 hours against Ae. aegypti were 1.7, 1.7, 1.8 ppm while LC₉₀ values were 3.7, 3.7 and 3.8 ppm respectively. Further no significant difference in LC₅₀ and LC₉₀ values of the formulation was observed against Ae. aegypti during 18 months storage period at room temperature. An application of the formulation at the rate of 140 mg a.i./m² in different breeding sites under natural field conditions provided 98.1% reduction of Anopheles larvae on day 1; thereafter 100% reduction was recorded up to week 1 and more than 80% reduction up to week 3, while percent reduction against Culex larvae was 95.5% on day 1, and thereafter 80% reduction was achieved up to week 3. The formulation also showed 95.1% and, 99.7% reduction of Aedes larvae on day 1 and day 2 respectively; thereafter 100% larval control was observed up to day 7.

Conclusion

The neem oil formulation was found effective in controlling mosquito larvae in different breeding sites under natural field conditions. As neem trees are widely distributed in India, their formulations may prove to be an effective and eco-friendly larvicide, which could be used as an alternative for malaria control.     

Larvicidal activity of some fungal extracts on Aedes caspius and Culex pipiens (Diptera: Culicidae)

A preliminary study was conducted to evaluate the effects of nine methanol fungal extracts on fourth instar larvae of Aedes caspius and Culex pipiens (Diptera: Culicidae). None of the extracts tested showed larvicidal activity except for the Paecilomyces lilacinus extract, which showed 100 % mortality against the larvae of Ae caspius after 24 h. The LC₅₀ values of the P. lilacinus extract after 24 h were 190.66 μg/mL against Ae. caspius and 254.25 μg/mL against Cx. pipiens, respectively. After 48 h, meanwhile, the LC₅₀ values were 65.70 and 164.13 μg/mL, respectively. Histological analysis of the midgut of the treated larvae of Ae. caspius and Cx. pipiens revealed changes such as cell destruction, spacing between the cells and disruption of the microvilli, resulting in an appearance of vacuolization in the midgut. This article is the first report of the use of P. lilacinus extract for the control of Ae. caspius and Cx. pipiens larvae and the data obtained may help to provide a better understanding of the mode of action of P. lilacinus as an insecticide against mosquito larvae.     

Repellent,larvicidal and adulticidal activities of essential oil from Dai medicinal plant Zingiber cassumunar against Aedes albopictus

Zingiber cassumunar is an important plant used in traditional medicine and as a natural mosquito repellent. However, the compounds responsible for the repellent activity of the plant are still unknown. The aim of the study is to identify the components of Z. cassumunar essential oil that show repellent activity against Aedes albopictus. We also evaluated the larvicidal and adulticidal activities of Z. cassumunar essential oil against Ae. albopictus. In-cage mosquito repellent experiments showed that Z. cassumunar essential oil possessed moderate repellent activity with a minimum effective dose (MED) of 0.16 ± 0.01 mg/cm², compared to reference standard N,N-diethyl-3-methylbenzamide (DEET, 0.03 ± 0.01 mg/cm²). Bioassay-guided fractionation identified the major active compound of Z. cassumunar essential oil as (−)-terpinen-4-ol (1) (MED: 0.19 ± 0 mg/cm²). We also found that Z. cassumunar essential oil showed moderate larvicidal activity against first instar larvae of Ae. albopictus with a LC₅₀ (50% lethal concentration) of 44.9 μg/L after 24 h. Fumigation bioassays showed that Z. cassumunar essential oil exhibits moderate adulticidal activity against Ae. albopictus with a LC₅₀ of 5.44%, while (−)-terpinen-4-ol showed significant adulticidal activity with a LC₅₀ of 2.10% after 24 h. This study verifies that the Z. cassumunar essential oil has mosquito repellent activity, and that (−)-terpinen-4-ol is mainly responsible for this activity. Furthermore, this study provides scientific support for the folk usage of Z. cassumunar essential oil as mosquito repellent and indicates that Z. cassumunar essential oil and (−)-terpinen-4-ol can be used as plant-derived repellents and insecticides for mosquito control.     

A recent survey of mosquito fauna in Guandong Province, southern China, with a review of past records

Abstract The southern province of Guangdong has long been subject to endemic mosquito‐borne diseases. In recent years, this region of China has experienced rapid, extensive economic development involving environmental change, making much of the scant knowledge of its mosquito fauna obsolete. This paper reviews previous mosquito surveys, some of which may be too old to be of relevance to present‐day conditions, and presents the results of a recent survey of adult and immature mosquitoes. The main vectors of mosquito‐borne diseases endemic to the area, such as dengue and Japanese encephalitis virus, develop in container habitats. A three‐year survey was carried out, between 2004 and 2006, of 4131 breeding containers in residential areas and in open, sparsely populated areas, of which approximately 50% were positive for the presence of mosquitoes, and 10 156 larvae and pupae were collected and identified. Twelve species were found in both residential and sparsely populated areas: Aedes albopictus (Skuse) (Diptera: Culicidae), Ae. lineatopennis (Ludlow), Ae. vexans (Meigen), Tanakaius togoi (Theobald), Culex barraudi Edwards, Cx dispectus Bram, Cx malayi (Leicester), Cx pallidothorax Theobald, Cx quinquefasciatus Say, Cx sitiens Wiedemann, Lutzia fuscanus Wiedemann and Tripteroides bambusa (Yamada). Armigeres subalbatus (Coquillett) was found only in containers in villages, whereas Ae. macfarlanei Edwards, Cx mimeticus Noé, Cx sinensis Theobald, Cx vegans Wiedemann, Cx wilfredi Colless and Mansonia uniformis (Theobald) were found only in non‐ or sparsely populated areas. In residential areas, the rank order of most common species, as measured by the proportion of containers colonized, was Ae. albopictus > Cx quinquefasciatus > Lu. fuscanus, whereas in sparsely populated areas the rank order was Cx quinquefasciatus > Ae. albopictus > Lu. fuscanus. Light traps in non‐ or sparsely populated areas caught 5995 adult mosquitoes of 25 species, some of which are not container breeders. The most common species were: Anopheles sinensis Wiedemann, An. maculatus Theobald, An. minimus Theobald, Ta. togoi, Cx bitaeniorhynchus Giles, Cx malayi, Cx quinquefasciatus, Cx sinensis Theobald, Cx sitiens, Cx tritaeniorhynchus Giles and Lu. fuscanus. It is noteworthy that nine species caught had not been previously recorded in Guangdong Province, highlighting the deficient knowledge of the current composition and distribution of the mosquito fauna of this part of China.     

Predator–prey interactions and the cannibalism of larvae of Armigeres subalbatus (Diptera: Culicidae)

Predation and cannibalism can affect the co-existence of mosquito species or the assemblage of mosquito species (i.e., community structure). In this study, predatory feeding patterns and cannibalism of larvae of Armigeres subalbatus mosquitoes were quantified under laboratory conditions using Aedes albopictus and Culex uniformis larvae as prey organisms. Rate of consumption of prey larvae and the rate of cannibalism of 1st to 4th instar larvae of Armigeres subalbatus were reported at 24 h intervals with and without alternative food supplement. Both the 3rd and 4th instar larvae of Ar. subalbatus showed a substantial predation on the larvae of Ae. albopictus (33.6 ± 4.4) and Cx uniformis (47.3 ± 7.6). The cannibalism of the predatory larvae was strongly correlated to the larval density (r = 0.9). The predator and the prey density and the given food type were significant factors that determined the rate of pupation, death and the emergence as adults. A significant relationship was shown for the co-occurrence of predatory larvae and the prey larvae at the 85 natural breeding habitats observed at the field (χ² = 74.4, p < 0.001). Nine breeding sites (10.6%) were positive for both prey and predatory larvae. Our study showed robust information about the predatory and cannibalistic behavior of Ar. subalbatus larvae emphasizing their role in managing the population structure of mosquito communities.     

Larvicidal effects of some essential oils against Aedes aegypti (L.), the vector of dengue fever in Saudi Arabia

Essential oils are very popular among organic growers because they are ecologically safe, do not have mammalian toxicity, and cannot be resistant to a variety of contaminants. Four essential oils, Lemon, Lavender, Peppermint, and Neem, were tested for larvicide efficacy against the dengue fever vector Aedes aegypti larvae under laboratory conditions using dipping bioassay techniques. Among the essential oils tested, lemon, peppermint, and lavender oils showed high larvicidal activity against larvae of Ae. aegypti. Lemon oil showed the highest effects (LC₅₀ 10.676 ppm), while Peppermint, Lavender and Neem oil showed the lowest effects (LC₅₀ 21.380, 29.818 and 38.058 ppm, respectively). As a result, the mixture of lemon oil (LC₅₀) with Peppermint oil (LC₂₅) showed the highest co-toxicity factor, whereas the mixture of Lemon oil (LC₅₀) with Diesel oil (LC₂₅) showed the lowest co-toxicity factor. Based on the results of this study, it appears that essential oils may be useful as larvicides against Ae. aegypti larvae. In search of new natural larvicides, these compounds may provide an alternative to Synthetic insecticides as these are environmentally safe insecticides.     

Oviposition substrate selection by Florida mosquitoes in response to pathogen‐infected conspecific larvae

The impact of the presence of larval mosquito pathogens with potential for biological control on oviposition choice was evaluated for three mosquito species/pathogen pairs present in Florida. These included Aedes aegypti infected with Edhazardia aedis, Aedes albopictus infected with Vavraia culicis, and Culex quinquefasciatus infected with Culex nigripalpus nucleopolyhedrovirus (CuniNPV). Two‐choice oviposition bioassays were performed on each host and pathogen species with one oviposition cup containing infected larvae and the other cup containing uninfected larvae (control). Both uninfected and E. aedis‐infected female Ae. aegypti laid significantly fewer eggs in oviposition cups containing infected larvae. Uninfected gravid female Ae. albopictus and Cx. quinquefasciatus oviposited equally in cups containing uninfected larvae or containing larvae infected with V. culicis or CuniNPV, respectively. Gravid female Ae. albopictus infected with V. culicis did not display ovarian development and did not lay eggs. The decreased oviposition by gravid Ae. aegypti in containers containing E. aedis‐infected larvae may indicate that the infected larvae produce chemicals deterring oviposition.     

Chicory (Cichorium intybus) and wormwood (Artemisia absinthium) extracts exhibit strong larvicidal activity against mosquito vectors of malaria,dengue fever,and filariasis

Vector-borne diseases transmitted by mosquitoes cause globally important diseases such as malaria, dengue fever, and filariasis. The incidence of these diseases can be reduced through mosquito control programs but these control programs currently rely on synthetic insecticides that can impact the environment, and has selected widespread mosquito resistance. Environment friendly and biodegradable natural insecticides discovered in plants offer an alternative approach to mosquito control. Here, we investigated extracts from root or aerial parts of Chicory (Cichorium intybus) and wormwood (Artemisia absinthium) against the early 4th instar larvae of Anopheles stephensi (malaria vector), Aedes aegypti (dengue fever vector), and Culex quinquefasciatus (filariasis vector). The root and aerial parts extracts of A. absinthium and C. intybus at 200, 100, 50, 25 and 12.5?ppm caused significant mortality of the tested mosquito species. Root extracts exhibited higher larvicidal activity that aerial part extracts. The highest larvicidal activity was recorded in methanol extract of roots of C. intybus with LC50?=?66.16, 18.88 and LC¬90?=?197.56, 107.16?ppm for An. stephensi; LC50?=?78.51, 40.15 and LC90?=?277.31, 231.28?ppm for Ae. aegypti and LC50?=?103.99, 64.56 and LC¬90?=?314.04, 247.54?ppm for Cx. quinquefasciatus. These results reveal potent mosquito larvicidal activity against vectors of malaria, dengue fever, and filariasis is present in extracts of chicory and wormwood.     

Biological activity of Xanthium strumarium seed extracts on different cancer cell lines and Aedes caspius,Culex pipiens (Diptera: Culicidae)

Effects of methanol extracts of Xanthium strumarium on different cancer cell lines and on the mortality rates of Aedes caspius, Culex pipiens (Diptera: Culicidae) were investigated. Among the cell lines tested, the Jurkat cell line was the most sensitive to the methanol extract and ethyl acetate fraction, with reported LC₅₀ values of 50.18 and 48.73 μg/ml respectively. Conversely, methanol extracts were not that toxic to the A549 cell line though the toxicity increased on further purification. The percentage of growth inhibition was dose dependent for the methanol extract and ethyl acetate fraction. The ethyl acetate fraction showed higher toxicity to all cell lines tested when compared to the methanol extract. The results showed that methanol extracts of plant seeds caused 100% mortality of mosquito larvae at a concentration of 1000 μg/ml after 24 h of treatment. The LC₅₀ and LC₉₀ values of X. strumarium were found to be 531.07 and 905.95 μg/ml against Ae. caspius and 502.32 and 867.63 μg/ml against Cx. Pipiens, respectively. From the investigations, it was concluded that the crude extract of X. strumarium showed a weak potential for controlling the larval instars of Ae. caspius and Cx. pipiens. However, on further purification the extract lost the larvicidal activity. The ethyl acetate fraction showed higher toxicity to all cell lines tested when compared to the methanol extract. The ethyl acetate fraction investigated in this study appears to have a weak larvicidal activity but a promising cytotoxic activity. Future studies will include purification and investigation in further detail of the action of X. strumarium on Cancer Cell Lines and mosquitoes.     

Development of permethrin resistance in Culex quinquefasciatus Say in Kuala Lumpur,Malaysia

The resistance status towards permethrin among the laboratory strain, the permethrin-selected strain and four field strains of Culex quinquefasciatus collected in Kuala Lumpur, Malaysia was determined using three standard laboratory methods: WHO larval bioassay, WHO adult bioassay and biochemical microplate assay. Cx. quinquefasciatus permethrin-selected strain larvae were the least susceptible to permethrin with a resistance ratio of 47.28-folds, whereas all field strain larvae of the same species were tolerant to permethrin with resistance ratios of more than 3-folds. In contrast, in adult stage, the permethrin exposed permethrin-selected strain (resistance ratio = 1.27) was found to be more susceptible to permethrin than all permethrin-exposed field strains (resistance ratios = 2.23–2.48). Complete mortalities for all strains of Cx. quinquefasciatus adults proved the effectiveness of the synergist; piperonyl butoxide (PBO). For the biochemical microplate assay, the reduction of the mean optical density of elevated oxidase activity of three field strains upon exposure to PBO confirmed the association between oxidase activity and permethrin tolerance. On the other hand, irregular patterns of the mean optical density of elevated oxidase activity in the laboratory strain, permethrin-selected strain and Jalan Fletcher strain illustrated the gene variation within these mosquito colonies as well as the involvement of other enzyme activities in the permethrin resistance occurred.     

Behavioral responses of Aedes aegypti,Aedes albopictus,Culex quinquefasciatus,and Anopheles minimus against various synthetic and natural repellent compounds

The behavioral responses of colony populations of Aedes aegypti, Aedes albopictus, Culex quinquefasciatus, and Anopheles minimus to four essential oils (citronella, hairy basil, catnip, and vetiver), two standard repellents (DEET and picaridin), and two synthetic pyrethroids (deltamethrin and permethrin) were conducted in the laboratory using an excito‐repellency test system. Results revealed that Cx. quinquefasciatus and An. minimus exhibited much stronger behavioral responses to all test compounds (65–98% escape for contact, 21.4–94.4% escape for non‐contact) compared to Ae. aegypti (3.7–72.2% escape (contact), 0–31.7% (non‐contact)) and Ae. albopictus (3.5–94.4% escape (contact), 11.2–63.7% (non‐contact)). In brief, essential oil from vetiver elicited the greatest irritant responses in Cx. quinquefasciatus (96.6%) and An. minimus (96.5%) compared to the other compounds tested. The synthetic pyrethroids caused a stronger contact irritant response (65–97.8% escape) than non‐contact repellents (0–50.8% escape for non‐contact) across all four mosquito species. Picaridin had the least effect on all mosquito species. Findings from the current study continue to support the screening of essential oils from various plant sources for protective properties against field mosquitoes.