Mosquitoes Larvicidal Activity of Ocimum kilimandscharicum Oil Formulation under Laboratory and Field-Simulated Conditions.

Mosquitoes are vectors of many severe diseases, including malaria, yellow as well as dengue fever, and lymphatic filariasis. The use of synthetic chemical insecticides for mosquito control has been associated with resistance development and detrimental human, and ecological effects. For a safer alternative, the emulsified Ocimum kilimandscharicum oil formulation was evaluated for its larvicidal activity. The oil was analyzed by GC and GC/MS. The formulations were evaluated against third instar mosquito larvae in the laboratory and later compared with Bacillus thuringiensis subsp. israelensis against An. gambiae under field-simulated conditions. Thirty-nine compounds were identified in the oil, the main ones being D-camphor (36.6%) and limonene (18.6%). The formulation showed significant larval mortalities against An. gambiae and An. arabiensis larvae with LC50 of 0.07 and 0.31 ppm, respectively, at 24 h. Under the field-simulated trial, within 24 h, the formulation showed 98% mortality while Bti had achieved 54%. On day three, it caused 100% mortality while Bti achieved 76.5%. The high bioactivity and sublethal toxic effects to offspring of treated mosquito larvae, in terms of disruption of larval morphological aspects, suggest the high potential of the formulation as a botanical larvicide. The formulation, thus, may provide a valuable alternative for the effective and eco-friendly control of disease vectors.

Growth-disrupting Murraya koenigii leaf extracts on Anopheles gambiae larvae and identification of associated candidate bioactive constituents.

Plant-based constituents have been proposed as eco-friendly alternatives to synthetic insecticides for control of mosquito vectors of malaria. In this study, we first screened the effects of methanolic leaf extracts of curry tree (Murraya koenigii) growing in tropical (Mombasa, Malindi) and semi-arid (Kibwezi, and Makindu) ecological zones of Kenya on third instar An. gambiae s.s. larvae. Extracts of the plant from the semi-arid region, and particularly from Kibwezi, led to high mortality of the larvae. Bioassay-guided fractionation of the methanolic extract of the leaves of the plants from Kibwezi was then undertaken and the most active fraction (20 fold more potent than the crude extract) was then analyzed by Liquid chromatography quadruple time of flight coupled with mass spectrometry (LC-QtoF-MS) and a number of constituents were identified, including a major alkaloid constituent, Neplanocin A (5). Exposure of the third instar larvae to a sub-lethal dose (4.43 ppm) of this fraction over 7-day periods induced gross morphogenetic abnormalities in the larvae, with reduced locomotion, and delayed pupation. Moreover, the few adults that emerged from some pupae failed to fly from the water surface, unlike in the untreated control group. These results demonstrate subtle growth-disrupting effects of the phytochemical blend from M. koenigii leaves on aquatic stages An. gambiae mosquito. The study lays down some useful groundwork for the downstream development of phytochemical blends that can be evaluated for integration into eco-friendly control of An. gambiae vector population targeting the often overlooked but important immature stages of the malaria vector.

An Assessment of Participatory Integrated Vector Management for Malaria Control in Kenya.

BACKGROUND: The World Health Organization (WHO) recommends integrated vector management (IVM) as a strategy to improve and sustain malaria vector control. However, this approach has not been widely adopted. OBJECTIVES: We comprehensively assessed experiences and findings on IVM in Kenya with a view to sharing lessons that might promote its wider application. METHODS: The assessment used information from a qualitative external evaluation of two malaria IVM projects implemented between 2006 and 2011 and an analysis of their accumulated entomological and malaria case data. The project sites were Malindi and Nyabondo, located in coastal and western Kenya, respectively. The assessment focused on implementation of five key elements of IVM: integration of vector control methods, evidence-based decision making, intersectoral collaboration, advocacy and social mobilization, and capacity building. RESULTS: IVM was more successfully implemented in Malindi than in Nyabondo owing to greater community participation and multistakeholder engagement. There was a significant decline in the proportion of malaria cases among children admitted to Malindi Hospital, from 23.7% in 2006 to 10.47% in 2011 (p < 0.001). However, the projects' operational research methodology did not allow statistical attribution of the decline in malaria and malaria vectors to specific IVM interventions or other factors. CONCLUSIONS: Sustaining IVM is likely to require strong participation and support from multiple actors, including community-based groups, non-governmental organizations, international and national research institutes, and various government ministries. A cluster-randomized controlled trial would be essential to quantify the effectiveness and impact of specific IVM interventions, alone or in combination. CITATION: Mutero CM, Mbogo C, Mwangangi J, Imbahale S, Kibe L, Orindi B, Girma M, Njui A, Lwande W, Affognon H, Gichuki C, Mukabana WR. 2015. An assessment of participatory integrated vector management for malaria control in Kenya. Environ Health Perspect 123:1145-1151; http://dx.doi.org/10.1289/ehp.1408748.

Zanthoxoaporphines A-C: Three new larvicidal dibenzo[de,g]quinolin-7-one alkaloids from Zanthoxylum paracanthum (Rutaceae).

The bioassay-guided purification of Zanthoxylum paracanthum (Rutaceae) extracts led to the isolation of three new alkaloids, namely 1-hydroxy-10-methoxy-7H-dibenzo[de,g]quinolin-7-one (zanthoxoaporphine A, 2), 1-hydroxy-7H-dibenzo[de,g]quinolin-7-one (zanthoxoaporphine B, 3) and 1,8-dihydroxy-9-methoxy-7H-dibenzo[de,g]quinolin-7-one (zanthoxaporphine C, 4), and a known lignan identified as sesamin (1). Isolation and purification of the constituent compounds was achieved through conventional chromatographic methods. The chemical structures of the isolated compounds were determined on the basis of UV, IR, NMR and MS data, and confirmed by comparison with those reported in the literature. The larvicidal activity of some of the isolated compounds was investigated by using third-instar Anopheles gambiae larvae.

Identification of insect-selective and mammal-selective toxins from Parabuthus leiosoma venom.

Venoms were collected from two scorpion species: Parabuthus leiosoma and Parabuthus pallidus from Kenya. Subcutaneous injection and oral toxicity tests of crude and pure fractions of scorpion venoms were done in Mus musculus (mice), Chilo partellus and Busseola fusca. The highest activity against C. partellus was found in P. leiosoma venom (LC(50) 0.689 mg/50mg body weight). Bioassay-guided purification by a combination of cation-exchange (CE) and reverse-phase high-performance liquid chromatography (RP-HPLC) led to the isolation of three toxic peptides. A lepidopteran-selective toxin (P. leiosoma insect toxin, Plit) was isolated, and the partial N-terminal amino acid sequence (-KDGYPVDNANCKYE-) plus the molecular weight (6688.5 Da) determined. A peptide with significant insect toxicity coupled with mild effects on mice (P. leiosoma toxin, Plt) was isolated, and the partial N-terminal amino acid sequence (-LCEKFKVQRLVELNCVD-) plus the molecular weight (6742.5 Da) was determined. Another toxin with anti-mammalian activity (P. leisoma mammal-selective toxin, Plmt), and N-terminal partial amino acid sequence of ADVPGNYPLDKNGNRYY- plus a molecular weight of 7145.5 Da was also isolated. Comparison of the partial N-terminal amino acid sequences with other toxins revealed that Plit shows high homology to other known insect toxins. Similarly, Plmt shows high homology with several birtoxin-like anti-mammalian toxins. Plt does not exhibit homology with any known scorpion toxin with combined anti-insect and anti-mammalian activity.

Repellency of essential oils of some Kenyan plants against Anopheles gambiae.

Essential oils of six plants growing in Kenya were screened for repellent activities against Anopheles gambiae sensu stricto. The oils of Conyza newii (Compositeae) and Plectranthus marrubioides (Labiateae) were the most repellent (RD50=8.9 x 10(-5) mg cm(-2), 95% CI) followed by Lippia javanica (Verbenaceae), Lippia ukambensis (Verbenaceae), Tetradenia riparia, (Iboza multiflora) (Labiateae) and Tarchonanthus camphoratus (Compositeae). Eight constituents of the different oils (perillyl alcohol, cis-verbenol, cis-carveol, geraniol, citronellal, perillaldehyde, caryophyllene oxide and a sesquiterpene alcohol) exhibited relatively high repellency. Four synthetic blends of the major components (present in > or = 1.5%) of the essential oils were found to exhibit comparable repellent activity to the parent oils.

New mosquito larvicidal tetranortriterpenoids from Turraea wakefieldii and Turraea floribunda.

The crude methanol extracts of the root barks of Turraea wakefieldii and Turraea floribunda were found to show mosquito larvicidal activity against third-instar larvae of Anopheles gambiae sensu stricto. Four new limonoids comprising a vilasininoid 1 and three havanensinoids 2-4 were isolated from the chloroform fractions of the methanol extracts of T. wakefieldii and T. floribunda, respectively. The structures of the compounds were elucidated by NMR spectroscopy. Compounds 1, 2, and 4 had LD50 values of 7.1, 4.0, and 3.6 ppm, respectively, and were more potent than azadirachtin, which had an LD50 value of 57.1 ppm when tested against larvae of A. gambiae.

Repellency of live potted plants against Anopheles gambiae from human baits in semi-field experimental huts.

The repellency of potted plants against the malaria vector Anopheles gambiae sensu stricto Giles was quantified in experimental huts under semi-field conditions inside a screen-walled greenhouse. Ocimum americanum Linnaeus (Labiatae), Lantana camara L. (Verbenaceae), and Lippia uckambensis Spreng (Verbenaceae) repelled at an average of 39.7% (95% confidence interval [CI] = 29.6-48.4%), 32.4% (95% CI = 19.7-43.1%), and 33.3% (95% CI = 21.5-43.3%) of the mosquitoes, respectively (P < 0.0001 for all treatments). This was determined by logistic regression, allowing for variations associated with different bait hosts, sampling huts, and replicate test nights. In contrast, Ocimum kilimandscharicum Guerke (Labiatae), Ocimum suave Willd. (Labiatae), Corymbia citriodora Hook (Myrtaceae), Azadirachta indica A. Juss (Meliaceae), Tagetes minuta L. (Asteraceae), and Hyptis suaveolens Poit. (Lamiaceae) did not significantly repel mosquitoes. The combination of O. americanum with either L. camara or L. uckambensis repelled 31.6% (95% CI = 19.7-41.7%) and 45.2% (95% CI = 34.7-54.0%) of the mosquitoes, respectively (P < 0.0001 for both treatments). This study is the first to show that live intact plants can reduce domestic exposure to malaria vector mosquitoes. As such, they may represent a new, sustainable and readily applicable malaria vector control tool for incorporation into integrated vector management programs.

Repellent activities of stereoisomers of p-menthane-3,8-diols against Anopheles gambiae (Diptera: Culicidae).

Four stereoisomers of p-menthane-3,8-diol, which make up the natural product obtained from Eucalyptus citriodora, were synthesized through stereoselective procedures. Repellency assays showed that all the four were equally active against Anopheles gambiae s.s. Racemic blends and the diastereoisomeric mixture of all the four isomers were also equally repellent. 1-alpha-terpeneol, with a single hydroxyl function at C-8 and unsaturation at C-8, and menthol, with a single hydroxyl function at C-3, were not repellent. The practical implication of these results is discussed.