Chemical composition and larvicidal activity of essential oils from Zingiber montanum (J. Koenig) Link ex. A. Dietr. against three mosquito vectors / Composición química y actividad larvicida de aceites esenciales de Zingiber montanum (J. Koenig) Link ex. A. Dietr. contra tres vectores de mosquitos

The chemical composition and larvicidal activity of essential oils derived from the leaves and rhizomes of Zingiber montanum (J. Koenig) Link ex. A. Dietr. were reported. The main compounds in the leaf oil were ß-pinene (13.8%), ß-phellandrene (11.3%) and α-pinene (7.3%) while the rhizome oil was dominated by sabinene (41.1%), terpinen-4-ol (22.7%) and (E)-nerolidol (14.3%). The minimum lethal concentration (larvicidal activity) LC50of the rhizome oil at 24 h against Aedes albopictus was 35.17 µg/mL, while LC50 values of 32.20 µg/mL and 31.12 µg/mL were obtained against Aedes aegypti and Culex quinquefasciatus respectively. At 48 h the oil displayed larvicidal action with LC50 values of 23.18 µg/mL, 25.58 µg/mL and 18.99 µg/mL respectively towards Ae. albopictus, Ae. Aegyptiand Cx. quinquefasciatus. The leaf oil did not exhibit significant mortality and larvicidal action. The results indicate the potential of rhizome essential oil of Z. montanumas a source of larvicidal agent.

En el presente trabajo se reportan la composición química y actividad larvicida de los aceites esenciales obtenidos de hojas y rizomas de Zingiber montanum (J. Koenig) Link ex. A. Dietr. Los principales compuestos en el aceite de hojas fueron ß-pineno (13.8%), ß-felandrene (11.3%) y α-pineno (7.3%); mientras que los más abundantes en el aceite de rizomas fueron sabineno (41.1%), terpinen-4-ol (22.7%) y (E)-nerolidol (14.3%). La concentración letal mínima (actividad larvicida) LC50 del aceite de riomas ante Aedes albopictus fue 35.17 µg/mL, mientras que los valores de LC50 de 32.20 µg/mL y 31.12 µg/mL fueron obtenidos ante Aedes aegyptiy Culex quinquefasciatus respectivamente. A las 48 horas, el aceite mostró acción larvicida con valores de LC50 de 23.18 µg/mL, 25.58 µg/mL y 18.99 µg/mL respectivamente, ante Ae. albopictus, Ae. Aegyptiand Cx. quinquefasciatus. El aceite de hojas no mostró mortalidad ni acción larvicida significativa. Los resultados indican el potencial del aceite esencial de rizomas de Z. montanum como una fuente de agentes larvicidas.

Isolation of mosquitocidal bacteria (Bacillus thuringiensis, B.sphaericus and B.cereus) from excreta of arid birds.

Mosquitocidal bacteria are environmentally friendly alternatives to chemical insecticides for controlling mosquitoes and therefore, there have been tremendous world-wide efforts to identify novel mosquitocidal bacteria from natural environment. In the present study, excreta from arid-birds were analyzed for identifying mosquitocidal bacteria. The selection of sample for bacterial screening is significant, because, arid-birds are the unique living species and gathering the foods from variety of sources from environment. Out of 1000 samples examined, twelve bacterial strains were identified as mosquitocidal and the 16S rRNA gene sequence alignment depicted that these isolates belonged to Bacillus species (Bacillus thuringiensis, B.sphaericus and B.cereus). Toxicity assay against mosquito vectors have shown that these isolates are potential. The B. sphaericus VCRC-B547 (NCBI: JN377789) has shown a higher toxicity against Cx. quinquefasciatus, An. stephensi, and Aed. aegypti. Result from SDS-PAGE has shown that there was considerable difference in the protein profiles among the new bacterial isolates. Phylogenetic tree with branch length 0.05 revealed three distinct groups with homology among the closely related Bacillus strains. This study therefore throws considerable interest on the diversity of microbial organisms from arid birds and its application in mosquito control.

Laboratory and field evaluation of two formulations of Bacillus thuringiensis M-H-14 against mosquito larvae in the Islamic Republic of Iran, 2012

Due to low efficacy in the field of a local commercial product of Bacillus thuringiensis M-H-14 [Bioflash[registered sign]], a study was designed to assess its efficacy in laboratory, glass standard aquarium, semi-field and field conditions at both target and higher dosages against immature stages of Anopheles spp. and Culex spp. In laboratory conditions, the LC[50] values of wettable powder and granule formulations were 227 and 1031 ppm respectively against a susceptible strain of An. stephensi. Following application of wettable powder and granules at 56.1 mg/aquarium, the survival rates of the exposed larvae to the granule formulation were 65.6% and 54.2% on days 6 and 8 respectively. In the artificial ponds, the larval density was reduced to 38.9%, 39.3% and 65.1% at dosages of 2, 4 and 8 g/m[2] respectively. In rice fields, at a dosage of 2 kg/ha, the density of immature larvae were reduced to 33.1% and 28.6% 7-days post-treatment. Further investigations are needed for the reasons for the low efficacy of this larvicide

Enhanced production of mosquitocidal cyclic lipopeptide from Bacillus subtilis subsp. subtilis.

Background & objectives: A cyclic lipopeptide, surfactin produced by a strain of Bacillus subtilis subsp. subtilis (VCRC B471) was found to exhibit activity against both the larval and pupal stages of mosquitoes. The present study was aimed at increasing the production of the mosquitocidal metabolite by modifying the conventional medium. Methods: Enhancement of mosquitocidal metabolite production was attempted by replacing the existing micronutrients of the conventional NYSM and supplementing the medium with additional amounts of glucose. The LC50 value of culture supernatant (CS) against the larval and pupal stages of Anopheles stephensi was determined. Crude mosquitocidal metabolite (CMM) was separated from the CS, identified by MALDI-TOF analysis and its LC50 dosage requirement for the pupal stage of the above mosquito species determined. Results: The medium containing a new composition of micronutrients and glucose up to 1 per cent resulted in increased metabolite production. The LC50 value of the CS obtained in the improved medium against larvae and pupae of An. stephensi was 5.57 and 0.71 μl/ml, respectively. The yield of CMM was doubled in the improved medium. MALDI-TOF analysis revealed that the CMM was surfactin. Interpretation & conclusions: The new improved medium enhanced the production of mosquitocidal metabolite as the dosage required for inciting 50 per cent mortality among the pupal stages of mosquitoes was only half of that required when the metabolite was produced in the conventional medium. The mosquitocidal metabolite was identified as surfactin, a cyclic lipopeptide and biosurfactant.

Adulticidal activity of essential oil of Lantana camara leaves against mosquitoes.

Background & objectives: Development of insect resistance to synthetic pesticides, high operational cost and environmental pollution have created the need for developing alternative approaches to control vector-borne diseases. In the present study we have investigated the insecticidal activity of essential oil isolated from the leaves of Lantana camara against mosquito vectors. Methods: Essential oil was isolated from the leaves of L. camara using hydro-distillation method. Bioassay test was carried out by WHO method for determination of adulticidal activity against mosquitoes. Different compounds were identified by gas chromatography-mass spectrometry analysis. Results: LD50 values of the oil were 0.06, 0.05, 0.05, 0.05 and 0.06 mg/cm2 while LD90 values were 0.10, 0.10, 0.09, 0.09 and 0.10 mg/cm2 against Ae. aegypti, Cx. quinquefasciatus, An. culicifacies, An. fluvialitis and An. stephensi respectively. KDT50 of the oil were 20, 18, 15, 12, and 14 min and KDT90 values were 35, 28 25, 18, 23 min against Ae. aegypti, Cx. quinquefasciatus, An. culicifacies, An. fluviatilis and An. stephensi, respectively on 0.208 mg/cm2 impregnated paper. Studies on persistence of essential oil of L. camara on impregnated paper revealed that it has more adulticidal activity for longer period at low storage temperature. Gas chromatographic-mass spectrometric analysis of essential oil showed 45 peaks. Caryophyllene (16.37%), eucalyptol (10.75%), α-humelene (8.22%) and germacrene (7.41%) were present in major amounts and contributed 42.75 per cent of the total constituents. Interpretation &conclusion: Essential oil from the leaves of L. camara possesses adulticidal activity against different mosquito species that could be utilized for development of oil-based insecticide as supplementary to synthetic insecticides.

Laboratory studies on mosquito larvicidal efficacy of aqueous & hexane extracts of dried fruit of Solanum nigrum Linn.

Background & objectives: Aqueous and organic solvent extracts of plants/plant parts were effective in killing the mosquito larvae. Comparative efficacy of the aqueous and hexane extracts of dried fruit of Solanum nigrum was tested against five laboratory colonized strains of mosquito species, namely Anopheles culicifacies species A, An. culicifacies species C, An. stephensi, Culex quinquefasciatus and Aedes aegypti to assess the possibility for use of these extracts for their control. Methods: Concentrations of aqueous extract of dried fruit in the range of 62.5 to 2000 ppm and hexane extract of dried fruit in the range of 0.781 to 150 ppm were used in bioassays. The mortality data were subjected to log probit regression analysis to detertmine the median lethal concentrations (LC50 and LC90) to kill 50 and 90 per cent of the treated larvae of the respective species. Results: All the five species registered 100 per cent mortality in larval bioassays at 1000 ppm with aqueous extract and at 100 ppm with hexane extract of dried fruit. In bioassays with aqueous extract An. culicifacies species A registered the lowest LC50 of 208.5 ppm (range-208.5-359 ppm for different mosquito species) while with hexane extract, An. stephensi registered the lowest LC50 of 6.25 ppm (6.25- 17.63 ppm for different mosquito species). The LC50 of aqueous extract was 13-39 fold higher than the values of hexane extract of dried fruit for different species. The calculated LC90 for hexane extract of dried fruit for different species was in the range of 43.38-95.28 ppm. Interpretation & conclusion: Hexane extract showed good mosquito larvicidal efficacy than that of the aqueous extract. The calculated LC 90 for the extract for different species was below 100 ppm and could be effective for comprehensive control of disease vectors.

Larvicidal activity of neem and karanja oil cakes against mosquito vectors, Culex quinquefasciatus (say), Aedes aegypti (L.) and Anopheles stephensi (L.).

Larvicidal effect of neem (Azadirachta indica) and karanja (Pongamia glabra) oil cakes (individuals and combination) was studied against mosquito species. Both the oil cakes showed larvicidal activity against the mosquito species tested. The combination of neem and karanja oil cakes in equal proportion proved to have better effect than the individual treatments. The combination of the two oil cakes recorded an LC95 of 0.93, 0.54 and 0.77% against the mosquitoes, Culex quinquefasciatus, Aedes aegypti and Anopheles stephensi respectively The increase in efficacy of the combination treatment over individuals in all the mosquito larvae tested was found to range about 4 to 10 fold in terms of LC50 and 2 to 6 fold in terms of LC95.

Control of mosquito vectors of tropical infectious diseases: (1) bioefficacy of mosquito coils containing several pyrethroids and a synergist.

The bioefficacy of mosquito coils containing several pyrethroids were tested in a 25 m3 room against Culex pipiens quinquefasciatus, Aedes aegypti and Anopheles dirus. The test results were compared with tests against Culex pipiens pallens in Japan. Based on the KT50 values (the 50% knockdown time) of mosquito coils containing dl, d-T80-allethrin, d, d-T-prallethrin and methoxymethyl-tetrafluorobenzyl tetramethyl-cyclopropanecarboxylate (K-3050) at doses of 0.05-0.5% (w/w) with or without a synergist, the pyrethroid susceptibility of the four mosquito species was as follows: Cx. p. quinquefasciatus was several times more tolerant to pyrethroids than Cx. p. pallens, Ae. aegypti was a further several times more tolerant than Cx. p. quinquefasciatus, and An. dirus was more susceptible than Cx. p. pallens (KT50 value: about half of Cx. p. pallens). The order of their susceptibilities is common for pyrethroids. Mosquito coils containing d, d-T-prallethrin and K-3050 at doses of 0.05-0.2% (w/w) and N-(2-ethylhexyl)bicycle-[2,2,1]-hept-5-ene-2,3-dicarboxyimide as a synergist at a ratio of 2 times the active ingredient were highly effective against Ae. aegypti, the most important mosquito vector for dengue fever.

Use of expanded polystyrene [EPS] and shredded waste polystyrene [SWAP] beads for control of mosquitoes

Mosquitoes transmit several diseases to human. There are several measures for control of larvae. As part of Integrated Vector Management [IVM] program, the utility of floating layers of polystyrene beads [EPS] is a potential alternative in habitats of mosquito larva. EPS beads prevent oviposition of mosquito as well as killing the immature stages by forming a tick layer on the water surface. They are cheap, environmentally safe and do not need frequent application and remain on the surface of water for long time. The objective of the current study was to asses the effectiveness of two types of polystyrene beads of [EPS] and [SWAP] for control of mosquito larvae under laboratory conditions. Anopheles stephensi and Culex quinquefasciatus were used for experimental purposes. In each tray 250 larvae of late 3rd and early 4th instars were introduced. The experiment was conducted on 4 replicates for An. stephensi, Cu. quinquefasciatus and combination of both. Emerging of adult mosquitoes were calculated every day until the end of experiments. Mortality rate and Inhibition of Emerge [IE] for Cu. quinquefasciatus, An. stephensi and combination of both species was 97.8%, 100% and 99.07%, respectively using EPS. In average, EPS was able to kill 98.9% of larvae. The figures with SWAP were 63%, 91.05% and 72.65%, respectively. The average mortality for mosquitoes was 75.57% EPS and SWAP beads can be very effective and practical for elimination of An. stephensi and Cx. quinquefasciatus under the laboratory conditions

Studies on mosquito larvicidal properties of Eucalyptus citriodora Hook (family-Myrtaceae).

Hexane extract obtained from leaves of Eucalyptus citriodora Hook was tested against larvae of Anopheles stephensi Liston, Culex quinquefasciatus Say and Aedes aegypti Linn to assess its toxicity and growth inhibiting activity. The results showed better activity of hexane extract against larvae of An. stephensi Liston as compared to the larvae of Cx. quinquefasciatus Say and Ae. aegypti Linn. The LC50 values against IVth instar larvae of three species were 69.86, 81.12 & 91.76 ppm respectively after 24 hours and 26.7, 29.9 & 38.8 ppm respectively after 72 hours. At lowest concentration viz. 10 ppm, 73% larvae of An. stephensi Liston failed to emergence as adult mosquito while in Cx. quinquefasciatus Say and Ae. aegypti Linn only 10 & 6% larvae failed to emerge. Further observations showed some morphological abnormalities in same pupae which could not moult normally. These results clearly indicated growth inhibition effect of the hexane extract of Eucalyptus against mosquito larvae, particularly Anopheles species. Thus Eucalyptus citriodora Hook, which is commonly found in Northern India, has a great potential for the development of an eco-friendly herbal insecticidal product for control of mosquito breeding.

Relative susceptibility of some common mosquito vector larvae to synthetic insecticidal compounds in north-western Rajasthan.

Relative susceptibility of three important mosquito vector larvae viz., Anopheles stephensi, Aedes aegypti and Culex quinquefasciatus, prevalent in the arid region was determined against four organophosphates (malathion, fenitrothion, fenthion, temephos) and three synthetic pyrethroid (alphamethrin, deltamethrin and fanvalerate) compounds. Studies were carried out on late 3rd or early 4th instar larvae of these species using standard WHO technique. Based on concentration mortality data LC50 and LC90 values along with their fiducial limits, regression equation, chi-square (chi2)/heterogeneity of the response have been determined by log probit regression analysis. LC50 values as observed for the above seven insecticides were 0.8097, 0.0398. 0.0432, 0.0035, 0.0025, 0.0092, 0.1006; 1.2370, 0.0531, 0.0655, 0.0076, 0.00004, 0.00004, 0.0046 and 1.4980, 0.0719, 0.0817, 0.0056, 0.00021, 0.00073, 0.0112 mg/l for the above three mosquito species respectively Among the four organophosphates tested temephos was the most effective followed by fenitrothion, fenthion and malathion. In general, Anopheles was found more susceptible as compared to the other two culicines to the above four compounds. The results also showed that larvae of Ae. aegypti were most susceptible followed by Cx. quinquefasciatus and An. stephensi to all the three pyrethroids tested. Among the three compounds tested alphamethrin was found to be the most toxic followed by deltamethrin while fanvalerate was the least toxic. The study would be of great importance while planning use of these insecticides for the control of different vector species in this area.

Chemical composition and larvicidal activities of the essential oil of Zanthoxylum armatum DC (Rutaceae) against three mosquito vectors.

BACKGROUND & OBJECTIVES: In view of the recently increased interest in developing plant origin insecticides as an alternative to chemical insecticide, this study was undertaken to assess the larvicidal potential of the essential oil from the seeds of Zanthoxylum armatum DC [syn. Z. alatum Roxb] (Rutaceae) against three medically important species of mosquito vectors, Aedes aegypti, Anopheles stephensi and Culex quinquefasciatus. METHODS: Essential oil was hydro distilled in the laboratory from the seeds obtained from the market and the chemical constituents of the oil were determined using GC/GC-MS. Bioefficacy of the essential oil was evaluated under laboratory conditions using III instar mosquito larvae. RESULTS: Among the three mosquito species tested, Cx. quinquefasciatus was the most sensitive (LC50 = 49 ppm) followed by Ae. aegypti (LC50 = 54 ppm) and An. stephensi (LC50 = 58 ppm). GC-MS analysis of the oil revealed at least 28 compounds, consisting mainly of oxygenated monoterpenes (75%) and monoterpenes (22%). Linalool though constituted a major part (57%), failed to produce any appreciable mortality when tested alone. INTERPRETATION & CONCLUSION: From the results it can be concluded that the larvae of the three mosquito species were susceptible to the essential oil composition. Such findings would be useful in promoting research aiming at the development of new agent for mosquito control based on bioactive chemical compounds from indigenous plant sources as an alternative to chemical larvicides.

Efficacy of piperonyl butoxide (PBO) as a synergist with deltamethrin on five species of mosquitoes.

Development of insecticide resistance has been a challenging problem for a long time and new solutions are yet to emerge. In this regard, the use of synergist with the insecticide is thought to play a key role in reducing the resistance levels. Present study demonstrates the efficacy of PBO with deltamethrin against the field collected mosquito larvae of five species of Aedes, Anopheles and Culexfrom in and around Mysore.

Repellency of essential oils extracted from plants in Thailand against four mosquito vectors (Diptera: Culicidae) and oviposition deterrent effects against Aedes aegypti (Diptera: Culicidae).

In this study we evaluated and reported repellent effects of essential oils from Thai plants against 4 mosquito vectors: Aedes aegypti, Ae. albopictus, Anopheles. dirus and Culex quinquefasciatus under laboratory conditions using human volunteers. The essential oils were extracted from 18 plant species, belonging to 11 families, and the oils were then prepared as 10% solution in absolute ethanol with additives. Two chemical repellents, deet and IR3535, were also prepared in the same formulation as the essential oil repellents and tested for repellency as controls. The essential oils were also evaluated for oviposition deterrent effects against Ae. aegypti under laboratory conditions. The results show night-biting mosquitoes (An. dirus and Cx. quinquefasciatus) and Ae. albopictus were more sensitive to all the essential oils (repellency 4.5 - 8 hours) than was Ae. aegypti (repellency 0.3 - 2.8 hours), whereas deet and IR3535 provided excellent repellency against all four mosquito species (repellency 6.7- 8 hours). All essential oils exhibited oviposition deterrent activity against Ae. aegypti with various degrees of repellency ranging from 16.6 to 94.7%, whereas deet and IR3535 had no repellency. The present study demonstrates the potential for using essential oils as mosquito repellents and oviposition deterrents. These findings may lead to new and more effective strategies for protection from and control of mosquitoes.

Sterculia guttata seeds extractives--an effective mosquito larvicide.

The larvicidal activity of ethanol, chloroform and hexane soxhlet extracts obtained from S. guttata seeds was investigated against the IVth instar larvae of Dengue fever vector, Aedes aegypti and filarial vector, Culex quinquefasciatus. All extracts including fractions of ethanol extract exhibited 100% larval kill within 24 hr exposure period at 500 ppm concentration. Fraction A1 of ethanol was found to be most promising; its LC50 was 21.552 and 35.520 ppm against C. quinquefasciatus and A. aegypti respectively. Naturally occurring S. guttata seed derived fractions merit further study as potential mosquito larval control agents or lead compounds.