Larvicidal effects of some Euro-Asiatic plants against Culex quinquefasciatus Say larvae (Diptera: Culicidae)

Methanol extracts of the aerial parts from 62 Euro-Asiatic plant species were tested for larvicidal activity against the mosquito Culex quinquefasciatus Say (Diptera: Culicidae) under laboratory conditions. The fourth larval instar was tested. The extracts of the plants Seseli pallasii and Schisandra chinencis displayed the highest larvicidal activities with LD₅₀ 6 and 15 ppm, respectively, followed by Arthemisia campestris, Verbena officinalis, and Imperatoria osthruthium with LD₅₀ 23, 38, and 49 ppm, respectively. The appraised value of LD₅₀ for two species was between 51–100 ppm; eight species had lethal doses from 101 to 500 ppm, 13 species showed lethal doses from 501 to 1,000 ppm, and 34 species did not show lethal doses for low mortality (LD₅₀ > 1,000 ppm).     

Insecticidal properties of essential oils from <Emphasis Type="Italic">Lippia turbinata</Emphasis> and <Emphasis Type="Italic">Lippia polystachya</Emphasis> (Verbenaceae) against <Emphasis Type="Italic">Culex quinquefasciatus</Emphasis> (Diptera: Culicidae)

Mosquitoes are important vectors of diseases to humans and domestic animals. Chemical control of vectors remains a main resource for the prevention and control of vector-borne diseases. Due to the development of insecticide resistance and risks to human health and the environment of synthetic compounds, the search for alternative pesticides is encouraged. This work assessed the insecticidal activity of essential oils (EOs) from Lippia turbinata and L. polystachya from Argentina on Culex quinquefasciatus mosquitoes. EOs were extracted by hydro-distillation and analyzed with gas–liquid chromatography and mass spectrometry. The insecticidal activity against mosquito larvae, pupae, and adults were evaluated according to World Health Organization protocols. Concentrations ranking from 10 to 160 ppm were assessed at 1, 2, 3, and 24 h posttreatment. The composition of the EO of L. polystachya and L. turbinata were qualitatively similar, with α-thujone and carvone as main constituent; differences were mostly due to the proportion of each component. β-caryophyllene was also an important constituent of the EO of L. turbinata. Both EO were larvicidal at concentrations of 80 ppm or higher, but only L. turbinata was adulticidal. No pupal mortality was detected. The potential of these EOs for vector control is discussed.     

Larvicidal and repellent activity of Hyptis suaveolens (Lamiaceae) essential oil against the mosquito Aedes albopictus Skuse (Diptera: Culicidae)

Lamiaceae have traditionally been used in developing countries for their insecticidal and repellent properties against several insect species. In our research, the essential oil (EO) extracted from fresh leaves of Hyptis suaveolens (Lamiaceae), and its main constituents were evaluated for larvicidal and repellent activity against the Asian tiger mosquito, Aedes albopictus Skuse (Diptera: Culicidae), currently the most invasive mosquito worldwide. H. suaveolens EO had insecticidal activity against A. albopictus larvae and mortality was dosage dependent. At the highest dosages of 450 and 400 ppm, there were no significant differences on larval mortality, as mortality ranged between 98.33% and 93.33%, respectively. At dosages ranging from 250 to 350 ppm, mortality rates were lower and not significantly different from each other. Terpinolene was found to be the most effective pure compound. Efficacy protection from H. suaveolens EO, at dosages ranging from 0.03748 to 0.7496 μg cm⁻² of skin, was evaluated during 150 min of observation. Results indicated that this EO had a significant repellent activity (RD₅₀ = 0.00035 μg cm⁻²; RD₉₀ = 0.00048 μg cm⁻²), with differences in repellency rates, as a function of both concentration and observation time. Protection time ranged from 16 to 135 min. These results clearly evidenced that the larvicidal and repellent activity of H. suaveolens EO could be used for the development of new and safer products against A. albopictus.     

Effects of essential oils from Cymbopogon citratus (DC) Stapf., Lippia sidoides Cham., and Ocimum gratissimum L. on growth and ultrastructure of Leishmania chagasi promastigotes

The current therapy for leishmaniasis, which affects annually about 2 million people, is far from satisfactory. All available drugs require parenteral administration and are potentially toxic. Plant essential oils have been traditionally used in folk medicine and appear as valuable alternative source for chemotherapeutic compounds. In this study, we demonstrated the effect of essential oils from Cymbopogon citratus, Lippia sidoides, and Ocimum gratissimum on growth and ultrastructure of Leishmania chagasi promastigote forms. Steam distillation was used to isolate the essential oils, and their constituents were characterized by gas chromatography coupled to mass spectrometry and nuclear magnetic resonance. All essential oils showed in vitro inhibitory action on L. chagasi promastigotes growth in a dose-dependent way, with IC₅₀/72 h of 45, 89, and 75 μg/mL for C. citratus, L. sidoides, and O. gratissimum, respectively. Drastic morphological alterations were observed in all essential oil-treated parasites, including cell swelling, accumulation of lipid droplets in the cytoplasm, and increase of acidocalcisome volume. Furthermore, aberrant-shaped cells with multi-septate body were observed by scanning electron microscopy, suggesting an additional effect on cytokinesis. Taken together, our data show that these essential oils affect the parasite viability being the C. citratus essential oil the most effective against L. chagasi.     

Evaluation of time toxicity, residual effect, and growth-inhibiting property of Carapa guianensis and Copaifera sp. in Aedes aegypti

Oils of Carapa guianensis and Copaifera spp. are well-known in the Amazonian region as natural insect repellents, and studies have reported their efficiency as larvicide against some mosquito species. However, toxicity persistence and effect on mosquito development have not yet been evaluated. Thus, the objective of this study was to evaluate the initial time of larvicidal activity, residual effect, and the effect of very low concentrations of these oils on Aedes aegypti. Different concentrations of the oils were used to evaluate the initial time of larval mortality and residual effect, as well as, the development of larvae, pupae, and adults. Results demonstrated that the lethal effect started mainly between the first 2 and 3 h of larvae exposure to oils, when using concentrations which ranged from 500 mg/L of C. guianensis and 90 mg/L of Copaifera sp. The toxic effect remained with total efficiency (100% mortality) until the sixth day for Copaifera sp. and 12th day for C. guianensis. When using sublethal dosages (ranging from 140 mg/L of C. guianensis to 26 mg/L of Copaifera sp.) mortality was observed after the larval molt. Also, imperfection of pupae and adult development and unsuccessful emergence of adults were observed. A product of botanical origin that could break the development of immature stage of mosquitoes and inhibit the emergence of adults should be essential in vector control. Thus, our results provide new information for a better understanding in using C. guianensis and Copaifera sp. oils with a potential to be used as a natural insecticide.     

Larvicidal potential of medicinal plant extracts against Anopheles subpictus Grassi and Culex tritaeniorhynchus Giles (Diptera: Culicidae)

Mosquito control is facing a threat due to the emergence of resistance to synthetic insecticides. Insecticides of botanical origin may serve as suitable alternative biocontrol techniques in the future. The acetone, chloroform, ethyl acetate, hexane, methanol and petroleum ether extracts of leaf, flower and seed of Cassia auriculata L., Leucas aspera (Willd.), Rhinacanthus nasutus KURZ., Solanum torvum Swartz and Vitex negundo Linn. were tested against fourth instar larvae of malaria vector, Anopheles subpictus Grassi and Japanese encephalitis vector, Culex tritaeniorhynchus Giles (Diptera: Culicidae). The larval mortality was observed after 24 h of exposure. All extracts showed moderate larvicidal effects; however, the highest mortality was found in leaf petroleum ether, flower methanol extracts of C. auriculata, flower methanol extracts of L. aspera and R. nasutus, leaf and seed methanol extracts of S. torvum and leaf hexane extract of V. negundo against the larvae of A. subpictus (LC₅₀ = 44.21, 44.69, 53.16, 41.07, 35.32, 28.90 and 44.40 ppm; LC₉₀ = 187.31, 188.29, 233.18, 142.66, 151.60, 121.05 and 192.11 ppm, respectively) and against the larvae of C. tritaeniorhynchus (LC₅₀ = 69.83, 51.29, 81.24, 71.79, 44.42, 84.47 and 65.35 ppm; LC₉₀ = 335.26, 245.63, 300.45, 361.83, 185.09, 351.41 and 302.42 ppm, respectively). These results suggest that the leaf petroleum ether, flower methanol extracts of C. auriculata, leaf and seed methanol extracts of S. torvum and leaf hexane extract of V. negundo have the potential to be used as an ideal eco-friendly approach for the control of the A. subpictus and C. tritaeniorhynchus. This is the first report on the mosquito larvicidal activity of the medicinal plant extracts.     

Evaluation of larvicidal nature of fleshy fruit wall of Momordica charantia Linn. (family: cucurbitaceae) in the management of mosquitoes

In view of the recently increased interest in developing plant-based insecticides as an alternative to chemical insecticides, this study was undertaken to assess the larvicidal potential of the various fruit wall extracts of Momordica charantia (cucurbitaceae) against two species of mosquito vectors, Anophels stephensi and Culex quinquefasciatus. Among the extracts tested, petroleum ether (LC₅₀ = 27.60; 17.22 ppm and 41.36; 15.62 ppm) extract was found more effective than carbon tetrachloride (LC₅₀ = 49.58; 16.15 ppm and 80.61; 27.64 ppm) and methanol (LC₅₀ = 142.82; 95.98 ppm and 1,057.49; 579.93 ppm) extracts towards anopheline and culicine larvae after 24 and 48 h of exposure respectively. Thus, all fruit wall extracts of M. charantia are toxic to both the larval species. M. charantia may, therefore, act as an effective biolarvicide against mosquitoes in the future.     

Biological activity of selected Lamiaceae and Zingiberaceae plant essential oils against the dengue vector Aedes aegypti L. (Diptera: Culicidae)

The larvicidal activity of hydrodistillate extracts from Mentha piperita L. Ocimum basilicum L. Curcuma longa L. and Zingiber officinale L. were investigated against the dengue vector Aedes aegypti L. (Diptera: Culicidae).The results indicated that the mortality rates at 80, 100, 200 and 400 ppm of M. piperita, Z. officinale, C. longa and O. basilicum concentrations were highest amongst all concentrations of the crude extracts tested against all the larval instars and pupae of A. aegypti. Result of log probit analysis (at 95% confidence level) revealed that lethal concentration LC₅₀ and LC₉₀ values were 47.54 and 86.54 ppm for M. piperita, 40.5 and 85.53 ppm for Z. officinale, 115.6 and 193.3 ppm for C. longa and 148.5 and 325.7 ppm for O. basilicum, respectively. All of the tested oils proved to have strong larvicidal activity (doses from 5 to 350 ppm) against A. aegypti fourth instars, with the most potent oil being M. piperita extract, followed by Z. officinale, C. longa and O. basilicum. In general, early instars were more susceptible than the late instars and pupae. The results achieved suggest that, in addition to their medicinal activities, Lamiaceae and Zingiberaceae plant extracts may also serve as a natural larvicidal agent.     

Chemical composition and larvicidal activity of essential oil from <Emphasis Type="Italic">Mentha spicata</Emphasis> (Linn.) against three mosquito species

Mosquitoes are blood-feeding insects and serve as the most important vectors for spreading human diseases such as malaria, yellow fever, dengue fever, and filariasis. The continued use of synthetic insecticides has resulted in resistance in mosquitoes. Synthetic insecticides are toxic and affect the environment by contaminating soil, water, and air, and then natural products may be an alternative to synthetic insecticides because they are effective, biodegradable, eco-friendly, and safe to environment. Botanical origin may serve as suitable alternative biocontrol techniques in the future. Mentha spicata, an edible and medicinal plant, is chiefly distributed in Southeast Asia and South Asia. In the present study, the toxicity of mosquito larvicidal activity of leaf essential oil (EO) and their major chemical constituents from Mentha spicata against Culex quinquefasciatus, Aedes aegypti, and Anopheles stephensi. The chemical composition of the leaf EO was analyzed using gas chromatography–mass spectroscopy (GC-MS). GC-MS revealed that the EO of M. spicata contained 18 compounds. The major chemical components identified were carvone (48.60%), cis-carveol (21.30%), and limonene (11.30%). The EO had a significant toxic effect against early third-stage larvae of C. quinquefasciatus, A. aegypti, and A. stephensi with LC₅₀ values of 62.62, 56.08, and 49.71 ppm and LC₉₀ values of 118.70, 110.28, and 100.99 ppm, respectively. The three major pure constituents extracted from the M. spicata leaf EO were also tested individually against three mosquito larvae. The LC₅₀ values of carvone, cis-carveol, and limonene appeared to be most effective against A. stephensi (LC₅₀ 19.33, 28.50, and 8.83 ppm) followed by A. aegypti (LC₅₀ 23.69, 32.88, and 12.01 ppm), and C. quinquefasciatus (LC₅₀ 25.47, 35.20, and 14.07 ppm). The results could be useful in search for newer, safer, and more effective natural larvicidal agents against C. quinquefasciatus, A. aegypti, and A. stephensi.     

Biochemical evidence of efficacy of potash alum for the control of dengue vector Aedes aegypti (Linnaeus)

Aedes aegypti is the primary vector of dengue, yellow fever and chikungunya in India and other South East Asian countries, and novel insecticides for vector control are urgently needed. In the present investigation, efficacy of potash alum, a traditionally known double salt in Indian and Chinese medicine system, was tested against the larvae of dengue vector, A. aegypti. LC_50, LC₉₀ and LC₉₉ values were recorded for various instar larvae where I instar larvae were found to be the most susceptible and IV instar larvae as the least susceptible one. The LC₅₀ values of crude and standard potash alum of various instar larvae ranged between 15.29 and 48.53 ppm and 20.50–65.10 ppm, respectively. Biochemical changes were also evidenced in IV instar A. aegypti larvae following a sublethal exposure for 24 h in the levels of various nutrient reserves and primary metabolites such as sugar, glycogen, lipids and proteins suggesting possible mode of action responsible for larval mortality. Sugar and glycogen concentrations were measured as 24.6 and 10.67 μg per five larvae in controls which were significantly (p < 0.05) reduced by 32.11–93.98% and 39.26–94.47%, respectively, in larvae treated with crude alum. In controls, protein and lipid content were recorded as 210.74 and 94.71 μg per five larvae which dropped up to 26.53% and 25.5%, respectively, in larvae following treatment with crude alum. Moreover, drastic changes were also recorded for DNA content with 25.39–44.17% decrease in crude alum-treated larvae. It is evident from these results that potash alum, a fairly cheaper and readily available ecofriendly compound could be recommended as a potential chemical larvicide against dengue vector at mosquito breeding sites in the vicinity of human dwellings.     

Larvicidal potential of ethanolic extracts of dried fruits of three species of peppercorns against different instars of an indian strain of dengue fever mosquito, Aedes aegypti L. (Diptera: Culicidae)

Larvicidal bioassay was carried out in the laboratory to assess the potential of ethanolic extracts of dried fruits of three species of peppercorns: Long pepper, Piper longum L., Black pepper, Piper nigrum, and White pepper, Piper nigrum against the different instars of field-collected Indian strain of dengue fever mosquito (Aedes aegypti L.). The investigations established the larvicidal potential of all the varieties of pepper fruits against Ae. aegypti. Against early fourth instar, the ethanolic extracts of Black and White P. nigrum proved to be 30–40% less toxic than the extracts of P. longum, whereas against third instars, white pepper extracts exhibited 7% more efficacy than that of black pepper and 47% more toxicity than that of long pepper. The results also revealed that the extracts of all the three pepper species were 11–25 times more toxic against the third instar larvae as compared to the early fourth instars. The lethal concentration, 50% (LC₅₀) values obtained with ethanolic extracts of P. longum, White P. nigrum and Black P. nigrum against early fourth instar larvae were 0.248, 0.356, and 0.405 ppm, respectively, and the lethal concentration, 90% (LC₉₀) values were 0.605, 0.758, and 0.801 ppm, respectively. Whereas against third instar larvae, the LC₅₀ values recorded with three extracts were 0.022, 0.015, and 0.016 ppm and the LC₉₀ values recorded were 0.054, 0.034, and 0.046 ppm, respectively. The larvae treated with all the pepper species showed initial abnormal behavior in their motion followed by excitation, convulsions, and paralysis, leading to 100% kill indicating delayed larval toxicity and effects of the extracts on the neuromuscular system. Observations of morphological alterations on treated larvae under light microscopy revealed that most organs, except anal papillae, had a normal structural appearance as that of controls. The structural deformation in the form of shrinkage in the internal membrane exhibited by anal papillae suggests the anal papillae as the probable action sites of the pepper extracts. The potential of peppercorns as new types of larvicides for the control of mosquitoes are explored.     

Larvicidal activity of lignans from Phryma leptostachya L. against Culex pipiens pallens

The larvicidal activity of crude petroleum ether, ethyl acetate, and methanol extracts of the whole plants of Phryma leptostachya L. was assayed for its toxicity against the early fourth instar larvae of Culex pipiens pallens. The larval mortality was observed after 24 h of exposure. Among three solvent extracts from Phyrma leptostachya L., the petroleum ether extract exhibited the best larvicidal activity. The corresponding LC₅₀ values of petroleum ether, ethyl acetate, and methanol extracts were 3.23, 5.23, and 61.86 ppm against the early fourth instar larvae of Culex pipiens pallens. The petroleum ether extract was successively subjected to column chromatography and preparative high performance liquid chromatography, and yielded the three lignans, phrymarolin-I, haedoxane A, and haedoxane E, which were isolated and identified as new mosquito larvicidal compounds. Phrymarolin-I, haedoxane A, and haedoxane E showed high larvicidal activity, for which the lethal doses LC₅₀ were estimated at 1.21, 0.025, and 0.15 ppm against the early fourth instar larvae of Culex pipiens pallens, respectively. The structures were elucidated by analyses of IR, UV, MS, and NMR spectral data. This is the first report on the mosquito larvicidal activity of the three compounds, phrymarolin-I, haedoxane A, and haedoxane E from Phyrma leptostachya L.     

Essential oil composition and larvicidal activity of six Mediterranean aromatic plants against the mosquito Aedes albopictus (Diptera: Culicidae)

Laboratory bioassays on insecticidal activity of essential oils (EOs) extracted from six Mediterranean plants (Achillea millefolium, Lavandula angustifolia, Helichrysum italicum, Foeniculum vulgare, Myrtus communis, and Rosmarinus officinalis) were carried out against the larvae of the Culicidae mosquito Aedes albopictus. The chemical composition of the six EOs was also investigated. Results from applications showed that all tested oils had insecticidal activity, with differences in mortality rates as a function of both oil and dosage. At the highest dosage (300 ppm), EOs from H. italicum, A. millefolium, and F. vulgare caused higher mortality than the other three oils, with mortality rates ranging from 98.3% to 100%. M. communis EO induced only 36.7% larval mortality at the highest dosage (300 ppm), a similar value to those recorded at the same dosage by using R. officinalis and L. angustifolia (51.7% and 55%, respectively). Identified compounds ranged from 91% to 99%. The analyzed EOs had higher content of monoterpenoids (80–99%) than sesquiterpenes (1–15%), and they can be categorized into three groups on the basis of their composition. Few EOs showed the hydrocarbon sesquiterpenes, and these volatile compounds were generally predominant in comparison with the oxygenated forms, which were detected in lower quantities only in H. italicum (1.80%) and in M. communis (1%).     

Repellence of essential oils of aromatic plants growing in Argentina against Aedes aegypti (Diptera: Culicidae)

Mosquitoes are vectors of pathogens to humans and domestic animals and may also have economical impacts. One approach to prevent mosquito-borne diseases is bite deterrence through the application of repellents. Currently, there is an interest to search for alternative bioactive products to the synthetic active ingredients most widely used in insect repellents. Repellence against Aedes aegypti of essential oils extracted from Acantholippia salsoloides, Aloysia catamarcensis, Aloysia polystachya, Lippia integrifolia, Lippia junelliana (Verbenaceae), Baccharis salicifolia, Euphatorium buniifolium, and Tagetes filifolia (Asteraceae) were assessed. Tests were conducted by alternatively exposing untreated and treated forehand to the mosquitoes and counting probing attempts. All essential oils tested were significantly repellent against A. aegypti when compared to untreated controls; L. junelliana was the most repellent and T. filifolia was the least based on the response of the mosquitoes to different concentrations of the essential oils (EO). Repellence may be attributed to the respective main components of each EO.     

Evaluation of indigenous plant extracts against the malarial vector, Anopheles stephensi (Liston) (Diptera: Culicidae)

Since ancient times, plant and microbial products were used in various aspects. However, their use against insects decreased when chemical products became developed. Recently, concerns increased with respect to public health and environmental security requiring detection of natural products that may be used against insects. In this study, mosquito Larvicidal and ovicidal activity of crude hexane, ethyl acetate, benzene, chloroform, and methanol extracts of the leaf of three plants, Eclipta alba, Cardiospermum halicacabum, and Andrographis paniculata, were tested against the early third-instar larvae of Anopheles stephensi (Liston) (Diptera: Culicidae). The larval mortality was observed after 24 h of exposure. All extracts showed moderate larvicidal effects; however, the highest larval mortality was found in methanol extract of A. paniculata, E. alba, and C. halicacabum against the larvae of A. stephensi (LC₅₀ = 79.68, 112.56, and 133.01 ppm; LC₉₀ = 154.66, 220.68, and 270.72 ppm), respectively. Mean percent hatchability of the ovicidal activity was observed 48 h post-treatment. The percent hatchability was inversely proportional to the concentration of extract and directly proportional to the eggs. Mortality of 100% with methanol and ethyl acetate extract of A. paniculata and methanol extract of E. alba were exerted at 200 ppm and methanol and benzene extract of C. halicacabum exerted at 150 ppm. This is an ideal eco-friendly approach for the control of the malaria vector, A. stephensi. Therefore, this study provides first report on the larvicidal and ovicidal activities against malaria vector, A. stephensi of E. alba plant extracts.     

Bioefficacy of larvicdial and pupicidal properties of Carica papaya (Caricaceae) leaf extract and bacterial insecticide, spinosad, against chikungunya vector, Aedes aegypti (Diptera: Culicidae)

The present study was carried out to establish the properties of Carica papaya leaf extract and bacterial insecticide, spinosad on larvicidal and pupicidal activity against the chikungunya vector, Aedes aegypti. The medicinal plants were collected from the area around Bharathiar University, Coimbatore, India. C. papaya leaf was washed with tap water and shade-dried at room temperature. An electrical blender powdered the dried plant materials (leaves). The powder (500 g) of the leaf was extracted with 1.5 l of organic solvents of methanol for 8 h using a Soxhlet apparatus and then filtered. The crude leaf extracts were evaporated to dryness in a rotary vacuum evaporator. The plant extract showed larvicidal and pupicidal effects after 24 h of exposure; however, the highest larval and pupal mortality was found in the leaf extract of methanol C. papaya against the first- to fourth-instar larvae and pupae of values LC₅₀ = I instar was 51.76 ppm, II instar was 61.87 ppm, III instar was 74.07 ppm, and IV instar was 82.18 ppm, and pupae was 440.65 ppm, respectively, and bacterial insecticide, spinosad against the first to fourth instar larvae and pupae of values LC₅₀ = I instar was 51.76 ppm, II instar was 61.87 ppm, III instar was 74.07 ppm, and IV instar was 82.18 ppm, and pupae was 93.44 ppm, respectively. Moreover, combined treatment of values of LC₅₀ = I instar was 55.77 ppm, II instar was 65.77 ppm, III instar was 76.36 ppm, and IV instar was 92.78 ppm, and pupae was 107.62 ppm, respectively. No mortality was observed in the control. The results that the leaves extract of C. papaya and bacterial insecticide, Spinosad is promising as good larvicidal and pupicidal properties of against chikungunya vector, A. aegypti. This is an ideal eco-friendly approach for the control of chikungunya vector, A. aegypti as target species of vector control programs.     

Larvicidal potential of silver nanoparticles synthesized using fungus Cochliobolus lunatus against Aedes aegypti (Linnaeus, 1762) and Anopheles stephensi Liston (Diptera; Culicidae)

Larvicides play a vital role in controlling mosquitoes in their breeding sites. The present study was carried out to establish the larvicidal activities of mycosynthesized silver nanoparticles (AgNPs) against vectors: Aedes aegypti and Anopheles stephensi responsible for diseases of public health importance. The AgNPs synthesized by filamentous fungus Cochliobolus lunatus, characterized by UV–Vis spectrophotometry, Fourier transform infrared spectroscopy, X-ray diffraction, scanning electron microscopy, and transmission electron microscopy. The characterization studies confirmed the spherical shape and size (3–21 nm) of silver nanoparticles. The efficacy of mycosynthesized AgNPs at all the tested concentrations (10, 5, 2.5, 1.25, 0.625, and 0.3125 ppm) against second, third, and fourth instar larvae of A. aegypti (LC₅₀ 1.29, 1.48, and 1.58; LC₉₀ 3.08, 3.33, and 3.41 ppm) and against A. stephensi (LC₅₀ 1.17, 1.30, and 1.41; LC₉₀ 2.99, 3.13, and 3.29 ppm) were observed, respectively. The mortality rates were positively correlated with the concentration of AgNPs. Significant (P < 0.05) changes in the larval mortality was also recorded between the period of exposure against fourth instar larvae of A. aegypti and A. stephensi. The possible larvicidal activity may be due to penetration of nanoparticles through membrane. Toxicity studies carried out against non-target fish species Poecilia reticulata, the most common organism in the habitats of A. aegypti and A. stephensi showed no toxicity at LC50 and LC90 doses of the AgNPs. This is the first report on mosquito larvicidal activity of mycosynthesized nanoparticles. Thus, the use of fungus C. lunatus to synthesize silver nanoparticles is a rapid, eco-friendly, and a single-step approach and the AgNps formed can be potential mosquito larvicidal agents.     

Efficacy of larvicidal botanical extracts against <Emphasis Type="Italic">Culex quinquefasciatus</Emphasis> Say (Diptera: Culicidae)

The present study explored the effects of crude leaf acetone, chloroform, hot water, methanol, petroleum ether (60–80°C), and water extracts of Calotropis procera (Ait) R. Br., Canna indica L., Hibiscus rosa-sinensis Linn., Ipomoea carnea Jacq. spp. fistulosa Choisy, and Sarcostemma brevistigma Wight that were selected for investigating larvicidal potential against second and fourth instar larvae of the laboratory-reared mosquito species, Culex quinquefasciatus Say, in which the major lymphatic filariasis was used. All plant extracts showed moderate larvicidal effects after 24 h of exposure at 1,000 ppm; however, the highest larval mortality was found in leaf acetone, chloroform, methanol, and petroleum ether of C. indica (LC₅₀ = 29.62, 59.18, 40.77, and 44.38 ppm; LC₉₀ = 148.55, 267.87, 165.00, and 171.91 ppm) against second instar larvae (LC₅₀ = 121.88, 118.25, 69.76, and 56.31 ppm; LC₉₀ = 624.35, 573.93, 304.27, and 248.24 ppm) and against fourth instar larvae and acetone, hot water, methanol, and petroleum ether extracts of I. carnea (LC₅₀ = 61.17, 41.07, 41.82, and 39.32 ppm; LC₉₀ = 252.91, 142.67, 423.76, and 176.39 ppm) against second instar larvae (LC₅₀ = 145.37, 58.00, 163.81, and 41.75 ppm; LC₉₀ = 573.30, 181.10, 627.38, and 162.63 ppm) and against fourth instar larvae of C. quinquefasciatus, respectively. These results suggest that the acetone, methanol extracts of C. indica and hot water, petroleum ether extracts of I. carnea have the potential to be used as an ideal eco-friendly approach for the control of the major lymphatic filariasis vector, C. quinquefasciatus.     

Larvicidal activities of Knema attenuata (Hook. f. & Thomson) Warb. (Myristicaceae) extracts against Aedes albopictus Skuse and Anopheles stephensi Liston

In recent years, uses of environment friendly and biodegradable natural insecticides of plant origin have received renewed attention as agents for vector control. The present study was undertaken to investigate the effect of aril and kernel extracts of Knema attenuata (Hook. f. & Thomson) Warb. (Myristicaceae) on larvae of Aedes albopictus Skuse and Anopheles stephensi Liston under laboratory conditions. The aril was extracted with chloroform and ethanol; the kernel was extracted with ethanol and hexane. The extracts were tested against the 3rd–4th instar larvae collected from Bunder area, Mangalore, India, which is a well-known fishing harbour, where several mosquito-borne diseases were reported. All the graded concentrations (100, 200, 300, 400 and 500 ppm) showed significant larval mortality after 24 h of observation. Chloroform extracts of aril showed 100% mortality against both larval forms of A. albopictus and A. stephensi at the concentration of 500 ppm. Among the extracts tested, chloroform extracts of aril and ethanol extracts of kernel exhibited higher toxicity against both A. albopictus (LC₅₀, 141 ppm and 159 ppm; LC₉₀, 290 ppm and 342 ppm) and A. stephensi (LC₅₀, 160 ppm and 162 ppm; LC90, 445 ppm and 458 ppm). Hexane extracts of kernel exhibited least toxicity against A. albopictus (LC50, 239 ppm; LC₉₀, 484 ppm), whereas ethanol extracts of aril showed the least toxicity against A. stephensi (LC₅₀, 290; LC₉₀, 498). A preliminary phytochemical assay revealed the presence of phenolics, tannins, steroids, terpenes, resins, and glycolipids in all the extracts. Alkaloids, flavonoids and saponins were absent. The lower LC₅₀ value of the chloroform extracts of K. attenuata aril indicates its potentiality as a larvicide against A. albopictus and A. stephensi mosquito larvae.     

Bioefficacy of Plumbago zeylanica (Plumbaginaceae) and Cestrum nocturnum (Solanaceae) plant extracts against Aedes aegypti (Diptera: Culicide) and nontarget fish Poecilia reticulata

In a search for natural products that could be used to control the vectors of tropical diseases, extracts of medicinal plants Plumbago zeylanica and Cestrum nocturnum have been tested for larvicidal activity against second, third, and fourth instar larvae of Aedes aegypti. The LC₅₀ values of all the extracts in different solvents of both the plants were less than 50 ppm (15.40–38.50 ppm) against all tested larval instars. Plant extracts also affected the life cycle of A. aegypti by inhibition of pupal development and adult emergence with increasing concentrations. The larvicidal stability of the extracts at five constant temperatures (19°C, 22°C, 25°C, 28°C, and 31°C) evaluated against fourth instar larvae revealed that toxicity of both plant extracts increases with increase in temperature. Toxicity studies carried out against fish species Poecilia reticulata, the most common nontarget organism in the habitats of A. aegypti, showed almost nil to meager toxicity at LC₅₀ and LC₉₀ doses of the plant extracts. The qualitative analysis of crude extracts of P. Zeylanica and C. nocturnum revealed the presence of bioactive phytochemicals with predominance of plumbagin in P. zeylanica and saponins in C. nocturnum. Partially purified plumbagin from P. zeylanica and saponins from C. nocturnum were obtained, and their presence was confirmed by thin-layer chromatography and biochemical tests. The bioassay experiment of partially purified secondary metabolites showed potent mosquito larvicidal activity against the fourth instar larval form. Therefore, this study explored the safer and effective potential of plant extracts against vector responsible for diseases of public health importance.