INSECTICIDAL ACTIVITY of NEEM, PYRETHRUM and QUASSIA EXTRACTS and THEIR MIXTURES AGAINST DIAMONDBACK MOTH LARVAE (Plutella Xylostella L.)
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MENDELNET 2016 INSECTICIDAL ACTIVITY OF NEEM, PYRETHRUM AND QUASSIA EXTRACTS AND THEIR MIXTURES AGAINST DIAMONDBACK MOTH LARVAE (Plutella xylostella L.) NAMBE JABABU, TOMAS KOPTA, ROBERT POKLUDA Department of Vegetable Science and Floriculture Mendel University in Brno Valticka 337, 69144 Lednice CZECH REPUBLIC [email protected] Abstract: The diamondback moth is known globally by many as the most destructive and economically important insect pest of cruciferous crops. It is also known to have developed resistance to numerous synthetic insecticides including those with newer active ingredients (Shelton et al. 2008); and this has triggered the development of alternative measures, including botanical insecticides (Oyedokun et al. 2011). The aim of this study was to evaluate the toxic effect of Azadirachta indica, Chrysanthemum cinerariaefolium and Quassia amara extracts and their mixtures on the diamondback moth larvae; in an attempt to find a superior mixture with more than one major active ingredient for the control of diamondback moth. The study consisted of two separate experiments; a feeding test, and a tarsal contact test. Mortality was recorded at 24, 48 and 72-h intervals from the start of the study. Varying levels of mortalities was recorded in both tests; mortalities ranged between 2–58% and 8% to 76% for the feeding test and the tarsal contact experiment respectively. In both experiments, the highest mortality was recorded in the first 24h and in formulations with the highest concentration. For the feeding test, Pyrethrins, Azadirachtin + Pyrethrins and Azadirachtin + Pyrethrins + Quassin extract/extract mixtures, produced the best effect; with Pyrethrins recording a 54% total mortality count, Azadirachtin + Pyrethrins combination producing a 48% mortality count and a 58% mortality count from Azadirachtin + Pyrethrins + Quassin extract combination. In the contact experiment, the highest mortality was observed in Pyrethrins, and Pyrethrins + Azadirachtin mixture; recording 76% and 64% mortality count respectively. Key Words: botanical insecticides, feeding test, contact test, larval mortality, brassica pests. INTRODUCTION The diamondback moth (Plutella xylostella) is considered the most destructive and economically important insect pest of cruciferous crops in the world (Sarfraz et al. 2006, You and Wei 2007). The economic loss due to this pest has been estimated worldwide to be US$4-US$5 billion (Zalucki et al. 2012). In addition to crop losses, the annual management costs for controlling this pest were estimated to be more than US$1.0 billion globally (Grzywaez et al. 2010). As a result, a lot of effort has been devoted to find alternative control measures for this pest because of the negative impact of pesticides and the problems encountered in controlling diamondback moth populations (Isman 2006). Having observed that some plants protect themselves better than others, humans developed the use of plants as pesticides. Historically, botanicals were used before other kinds of pesticides. They are mentioned in Hieroglyph, Chinese, Greek, and Roman antiquity and also in India where the use of the neem tree (Azadirachta indica Juss.; Meliaceae) was reported in the Veda, a body of manuscripts written in archaic Sanskrit dated at least 4,000 years ago (Philogène et al. 2005). Botanical pesticides are naturally occurring chemicals extracted from plants. The plant kingdom is the most efficient producer of chemical compounds (primary and secondary metabolites), synthesizing many products having wide array of functions that are used in defence against herbivores (Croteau et al. 2000). Extracts from Azadirachta indica, Chrysanthemum cinerariifolium and Quassia amara have all been reported individually to possess insecticidal effects against the diamondback moth larvae. Grainge et al. 1984, and others; stated the neem plant possessed Insecticidal, Contact Poison, Stomach Poison, 84 | Page MENDELNET 2016 Growth Inhibitory, Antifeedant and Repellant activity against the diamondback moth. They also reported that the whole plant, bark, stem, leaves, fruits and seeds, were among plant parts from which extracts could be made. Pyrethrum (active component being pyrethrins) is the powdered, dried flower head of the daisy; Chrysanthemum cinerariaefolium (Asteraceae). The flowers are ground to a powder and then extracted with hexane or a similar nonpolar solvent (Casida and Quistad 1995, Glynne-Jones 2001). The insecticidal action of the pyrethrins is characterized by a rapid knockdown effect, particularly in flying insects, and hyperactivity and convulsions in most insects. These symptoms are a result of the neurotoxic action of the pyrethrins, which block voltage-gated sodium channels in nerve axons. In purity, pyrethrins are moderately toxic to mammals (rat oral acute LD50 values range from 350 to 500 mg/kg), but technical grade pyrethrum is considerably less toxic (ca. 1,500 mg/kg) (Casida and Quistad 1995). Technical grade pyrethrum, the resin used in formulating commercial pesticides, typically contains from 20–25% pyrethrins (Casida and Quistad 1995, Isman 2006). Two types of botanical pesticides can be obtained from seeds of the Indian neem tree; Azadirachta indica (Schmutterer 1990, 2002); at the physiological level, azadirachtin blocks the synthesis and release of molting hormones from the prothoracic gland, leading to incomplete ecdysis in immature insects. And in adult female insects, a similar mechanism of action leads to sterility. In addition, azadirachtin is a potent antifeedant to many insects. Neem oil, obtained by cold-pressing seeds, can be effective against soft-bodied insects and mites but is also useful in the management of phytopathogens. Apart from the physical effects of neem oil on pests and fungi, disulfides in the oil likely contribute to the bioactivity of this material (Dimetry 2012). Quassia amara is a neotropical forest shrub or small tree, whose range extends from Mexico to Ecuador, including the Caribbean basin, where it normally grows in the forest understory, but it also grows easily in disturbed areas (Villalobos 1995). Its wood contains several quassinoids with insecticidal properties (Polonsky 1973). Daido et al. (1995), also noted that many cytotoxic quassinoids have been shown to have insect antifeedant activity. It is assumed that a binary mixture (having two components, A and B) can elicit at least three types of responses on an insect (Bitterman 1996, Caouvillon and Bitterman 1982). Two of these correspond to each of the components, where the response of one of the components dominates over the response to the other, and would be similar to the effects produced when presented separately. These elemental qualities might be diminished or enhanced in intensity via mixture suppression or synergism (Cromarty and Derby 1997). A mixture of A and B could also give rise to a third element (AB) that is activated only when both elemental stimuli are present. And it is AB that is associated with reinforcement instead of A and B themselves (Rudy and Sutherland, 1992). Based on the many available evidence on the insecticidal activities of the Neem, Pyrethrum and the Quassia amara extracts individually, we decided to further investigate the effects of extracts from those plant; individually and in a 1:1 and 1:1:1 mixture on DBM larvae under laboratory conditions. MATERIALS AND METHODS This research work was conducted in the laboratories of the Faculty of Horticulture of the Mendel University in Brno (Czech Republic) between September and December 2015. All botanical pesticides (NeemAzal T/S, Spruzit®, and Quassia wood chips) used for this experiment were purchased from local authorized dealers in the Czech Republic. Cabbage, Brassica oleracea var. capitata L. (HORNET F1) was planted in plastic pots (100 mm diameter) in a mixture of 70% peat substrate and 20% perlite. Plants were maintained under normal greenhouse conditions, and six weeks-old plants were used for the experiments. Plutella xylostella larvae used in this study were obtained from the laboratory Crop Research Institute; Czech Republic. The colony was reared on cabbage seedlings (Brassica oleracea var capitata L.) and maintained at room temperature (20–25 °C), RH of 60–65% and a 16:8 h light: dark regime. Bio-pesticides Used A test solution of Spruzit, [with 4.59 g/l Pyrethrins; equivalent to 18.36 g/l natural pyrethrum and 825.3 g/l rapeseed oil]; with different concentrations (2.0%, 1.0%, and 0.50% v/ v) was prepared with 85 | Page MENDELNET 2016 PYRETHRIN being the active ingredient. For the Neem extract/Azadirachtin (NeemAzal-T/S), with azadirachtin 10.6 g/l (Azadirachtin A 1%); a test solution with different concentrations of NeemAzal- T/S (2.0%, 1.0%, and 0.5% v/ v) were prepared with AZADIRACHTIN being the active ingredient. A test solution of 3% w/v which was latter diluted to 1.5% and 0.75%, was also prepared from Quassia amara (Quassia wood chips) with QUASSIN being the active component. The aqueous extract of Quassia amara was prepared by soaking Quassia wood chips overnight in water and boiled for 30 minutes, following procedures similar to those described by Dodia et al. (2008) and Zijp and Blommers (2002). A brown-coloured solution which was subsequently obtained was decanted and used immediately. The supplier did however not specify the amount of quassins in the wood chips, but it is known that the source of quassin and neoquassin is the wood of Quassia amara L. (Sapindales: Simaroubaceae), containing; depending on the age,