Alfred Elbert

Overview of the Status and Global Strategy for Neonicotinoids

In recent years, neonicotinoid insecticides have been the fastest growing class of insecticides in modern crop protection, with widespread use against a broad spectrum of sucking and certain chewing pests. As potent agonists, they act selectively on insect nicotinic acetylcholine receptors (nAChRs), their molecular target site. The discovery of neonicotinoids can be considered as a milestone in insecticide research and greatly facilitates the understanding of functional properties of the insect nAChRs. In this context, the crystal structure of the acetylcholine-binding proteins provides the theoretical foundation for designing homology models of the corresponding receptor ligand binding domains within the nAChRs, a useful basis for virtual screening of chemical libraries and rational design of novel insecticides acting on these practically relevant channels. Because of the relatively low risk for nontarget organisms and the environment, the high target specificity of neonicotinoid insecticides, and their versatility in application methods, this important class has to be maintained globally for integrated pest management strategies and insect resistance management programs. Innovative concepts for life-cycle management, jointly with the introduction of generic products, have made neonicotinoids the most important chemical class for the insecticide market.

Applied aspects of neonicotinoid uses in crop protection.

Neonicotinoid insecticides comprise seven commercially marketed active ingredients: imidacloprid, acetamiprid, nitenpyram, thiamethoxam, thiacloprid, clothianidin and dinotefuran. The technical profiles and main differences between neonicotinoid insecticides, including their spectrum of efficacy, are described: use for vector control, systemic properties and versatile application forms, especially seed treatment. New formulations have been developed to optimize the bioavailability of neonicotinoids through improved rain fastness, better retention and spreading of the spray deposit on the leaf surface, combined with higher leaf penetration. Combined formulations with pyrethroids and other insecticides are also being developed with the aim of broadening the insecticidal spectrum of neonicotinoids and to replace WHO Class I products from older chemical classes. These innovative developments for life-cycle management, jointly with the introduction of generic products, will, within the next few years, turn neonicotinoids into the most important chemical class in crop protection.

Resistance of Bemisia tabaci (Homoptera: Aleyrodidae) to insecticides in southern Spain with special reference to neonicotinoids

The tobacco whitefly, Bemisia tabaci Gennadius (Homoptera: Aleyrodidae) which occurs in various parts of the world, has developed a high degree of resistance against several chemical classes of insecticide, including organophosphates, carbamates, pyrethroids, insect growth regulators and chlorinated hydrocarbons. The present studies were done in order to monitor the susceptibility of whitefly populations in southern Spain to insecticides commonly used there. Systemic bioassays using Spanish field populations of B tabaci collected in 1994, 1996 and 1998 indicated an increase, albeit a slow one, in resistance to imidacloprid over this period. Comparative studies of other neonicotinoids using the same bioassay revealed a high degree of cross-resistance to acetamiprid and thiamethoxam. Leaf-dip bioassays with adult females from these populations revealed a high level of resistance to cyfluthrin, endosulfan, monocrotophos, methamidophos, and pymetrozine, each at 200 mg litre−1. Buprofezin and pyriproxyfen were tested against second-instar nymphs and eggs, respectively. Buprofezin also showed a lower efficacy against ESP-98, a strain of B tabaci received from Almeria in 1998, but pyriproxyfen resistance was not obvious when tested against eggs of strain ESP-98. Field trials in 1998 revealed good efficacy of imidacloprid in one farm in the Almeria region and two greenhouses in Murcia and Sevilla, but a loss of activity by imidacloprid in another farm in the Almeria region. Cross-resistance between imidacloprid and thiamethoxam was also confirmed under field conditions. © 2000 Society of Chemical Industry

Baseline determination and detection of resistance to imidacloprid in Bemisia tabaci (Homoptera: Aleyrodidae)

Adult Bemisia tabaci (Gennadius) were tested with imidacloprid in a bioassay method using systemically treated cotton leaves. The method was simple, robust and repeatable and provided baseline data for a laboratory and a number of recently collected field strains. The LC 50 for imidacloprid susceptible strains was calculated to be 1.7 ppm and a concentration of 16 ppm determined as diagnostic for imidacloprid resistance. Ten strains of B. tabaci collected from the Almeria region of Spain showed significantly less mortality at the diagnostic dose than the susceptible strains. The intensive use of imidacloprid in the Almeria region is considered the reason for the occurrence of resistance in this locality. Resistance to organophosphates, pyrethroids and endosulfan in B-type and non-B type B. tabaci did not confer resistance to imidacloprid.

Efficacy of plant metabolites of imidacloprid against Myzus persicae and Aphis gossypii (Homoptera: Aphididae)

The metabolism of the chloronicotinyl insecticide imidacloprid is strongly influenced by the method of application. Whilst in foliar application most of the residues on the leaf surface display unchanged parent compound, most of the imidacloprid administered to plants by soil application or seed treatment is metabolized more or less completely, depending on plant species and time. The present study revealed that certain metabolites of imidacloprid which have been described in crop plants are highly active against aphid pests in different types of bioassays. Some of these metabolites showed a high oral activity against the green peach aphid (Myzus persicae), and the cotton aphid (Aphis gossypii). The aphicidal potency of the metabolites investigated was weaker in aphid dip tests than in oral ingestion bioassays using artificial double membranes. The most active plant metabolite was the imidazoline derivative of imidacloprid. The LC50 values of this metabolite for M. persicae and A. gossypii in oral ingestion bioassays were in the lower ppb-range, i.e. 0·0044 and 0·0068 mg litre-1, respectively. Most of the other reported metabolites showed much weaker activity. Compared to imidacloprid, the imidazoline derivative showed superior affinity to housefly (Musca domestica) head nicotinic acetylcholine receptors, while all other metabolites were less specific than imidacloprid. It seems possible that, after seed treatment or soil application, a few of the biologically active metabolites arising are acting in concert with remaining levels of the parent compound imidacloprid, thus providing good control and long-lasting residual activity against plant-sucking pests in certain crops.

Imidacloprid, a Novel Chloronicotinyl Insecticide: Biological Activity and Agricultural Importance

Following the discovery of the insecticidal properties of the heterocyclic nitromethylenes (Soloway et al. 1978), chemists of Nihon Bayer Agrochem started in 1979 to optimize these structures. In 1985 the coupling of the chloropyridyl moiety to the N-nitro substituted imidazolidine ring system enabled the synthesis of the highly active insecticide imidacloprid (Fig. 1). Imidacloprid is the first commercial example of the chloronicotinyl insecticides acting on nicotinic acetylcholine receptors (Leicht 1993). It is now registered in more than 60 countries as a compound with a new or non-conventional mode of action to combat highly resistant insect pests (Elbert et al. 1991; Elbert et al. 1996; Nauen et al. 1996a). Chloronicotinyl insecticides will grow in importance in the coming years because other close analogues of imidacloprid, such as Takeda’s and Nippon Soda’s open chain derivatives nitenpyram and acetamiprid, respectively, have been described (Tomizawa et al. 1995; Yamamoto et al. 1995). During recent years several studies have demonstrated the excellent activity of imidacloprid on pest species of different orders. The present chapter gives an overview of the biological activity of imidacloprid on different target pests, its selectivity even at the molecular level, its physicochemical properties which led to good systemicity and its agricultural importance.

Whitefly‐active metabolites of imidacloprid: biological efficacy and translocation in cotton plants

The chloronicotinyl insecticide imidacloprid is widely used in soil application, seed treatment and as a foliar spray. Its systemic properties are well-known. It is more or less completely metabolised, depending on the method of application, plant species and time. Some of the metabolites that arise are active against different aphid species, as shown earlier, with the imidazoline derivative (olefine metabolite) more active in oral ingestion bioassays than the parent compound itself. In the present work, we demonstrate that the olefine metabolite and two hydroxy metabolites of imidacloprid are also active against the cotton whitefly, Bemisia tabaci, in oral ingestion bioassays (sachet test). The 4-hydroxy metabolite is as active as imidacloprid and the olefine compound c10 times more active. The two hydroxy metabolites were also active against biotypes from Almeria, Spain and a B-type strain from California. The physicochemical properties of the metabolites were determined and compared with those of imidacloprid. Lower log P values were found for all metabolites investigated. Interestingly, the olefine metabolite was the only compound with an acidic proton (pKa 7.2) under neutral pH conditions, suggesting that it could be more phloem-mobile. In order to investigate this hypothesis we compared the translocation of the radiolabelled olefine metabolite after foliar application with the movement of radiolabelled imidacloprid in cotton plants, but no major differences were found in acropetal or basipetal translocation.

Antifeedant effects of sublethal dosages of imidacloprid on Bemisia tabaci

The effects of sublethal dosages of the chloronicotinyl insecticide imidacloprid on different strains of the tobacco whitefly, Bemisia tabaci Gennadius (Homoptera: Aleyrodidae), have been studied after leaf dip and systemic application. All bioassays were performed with the insecticide susceptible strain, SUD‐S, and two Spanish biotypes, ALM‐2 and LMPA‐2, both resistant to conventional insecticides and with a lower susceptibility towards imidacloprid. Honeydew, excreted by all strains feeding on treated and untreated cotton leaf discs was quantified by photometric analysis of its carbohydrate content. EC50‐values for the depression of honeydew excretion in female adults after systemic application of imidacloprid were calculated at 0.037 ppm, 0.027 ppm and 0.048 ppm for strains SUD‐S, ALM‐2 and LMPA‐2, respectively, indicating no significant differences between strains in feeding behaviour throughout an 48 h testing period. Depending on the strain these EC50‐values were 150‐ to 850‐times lower than LC50‐values calculated for mortality in the same bioassay. Starvation tests revealed mean survival times of >48 h for female adults placed on agar without leaf discs, indicating that sublethal dosages of imidacloprid which caused antifeedant responses, were probably not covered in common 48 h systemic bioassays, used to monitor resistance to imidacloprid. Effects of sublethal dosages on honeydew excretion after leaf dip application seem to be minor. In choice situations with systemically treated and untreated leaf discs in a single container, female adults of B. tabaci showed a clear preference for the untreated leaf discs. However, when using leaf discs treated by painting the surface with imidacloprid in the same bioassay, feeding activities on treated and untreated leaf discs were not significantly different. The results of the present study demonstrate the antifeedant properties of imidacloprid on B. tabaci, which might play an essential role after soil application or seed treatment under field conditions.

Aphicidal activity of imidacloprid against a tobacco feeding strain of Myzus persicae (Homoptera: Aphididae) from Japan closely related to Myzus nicotianae and highly resistant to carbamates and organophosphates

We investigated the resistance potential of a red-coloured Japanese strain (JR) of a tobacco feeding form of Myzus persicae (Sulzer) of the M. persicae species complex closely related to the tobacco aphid Myzus nicotianae Blackman. Bioassays were performed with a range of insecticides, imidacloprid, nicotine and cartap, thought to act on nicotinic acetylcholine receptors in vivo , as well as with two conventional insecticides, pirimicarb and oxydemeton-methyl, acting on acetylchol-inesterase (AChE). Compared to a susceptible strain, JR showed high resistance to pirimicarb and oxydemeton-methyl, but was far less resistant to nicotine, cartap and imidacloprid. Imidacloprid was, among the insecticides tested, the most active compound in contact and ingestion bioassays. Compared to the susceptible strain, JR showed four-to seven-fold resistance to imidacloprid depending on the type of bioassay. Resistance factors for other insecticides tested in an oral ingestion bioassay were: cartap five-fold, nicotine nine-fold, oxydemeton-methyl 107-fold and pirimicarb > 385-fold. JR showed high carboxylesterase activity. Polyacrylamide gel electrophoresis indicated esterase FE4 as the major carboxylesterase. As for most M. persicae strains and some Greek strains of M. nicotianae , JR was monomorphic for glutamate oxalacetate transaminase. Studies with pirimicarb showed a marked insensitivity of AChE to inhibition by this chemical, whilst such insensitivity could not be detected with the organophosphate insecticide oxydemeton-methyl. Receptor binding assays with [ 3 H]-imidacloprid in aphid homogenates revealed I 50 -values of 0.4 to 0.8 nM and no statistical difference between the JR and susceptible strain.

European monitoring of resistance to insecticides in Myzus persicae and Aphis gossypii (Hemiptera: Aphididae) with special reference to imidacloprid.

The susceptibility to several insecticides of 16 and 8 strains of Myzus persicae Sulzer and Aphis gossypii Glover, respectively, received from different European countries in 2001 was investigated. Most of the strains were derived from places known for their aphid resistance problems to conventional insecticides before imidacloprid was introduced. In many regions and agronomic cropping systems imidacloprid has been an essential part of aphid control strategies for a decade, and therefore the susceptibility of aphid populations to imidacloprid using FAO-dip tests and diagnostic concentrations in a leaf-dip bioassay was checked. Additional insecticides tested were cyfluthrin (chemical class: pyrethroid), pirimicarb (carbamate), methamidophos and oxydemeton-methyl (organophosphates). Diagnostic concentrations (LC99-values of reference strains) for each insecticide were established by dose response analysis using a new leaf-disc dip bioassay format in 6-well tissue culture plates. Virtually no resistance to imidacloprid in any of the field-derived populations of M. persicae and A. gossypii was detected. In contrast, strong resistance was found to pirimicarb and oxydemeton-methyl, and to a lesser extent also to cyfluthrin. Two strains of A. gossypii exhibited reduced susceptibility to imidacloprid when tested directly after collection. However, after maintaining them for six weeks in the laboratory, the aphids were as susceptible as the reference strain. The diagnostic concentration of methamidophos did not reveal any resistance in M. persicae, but did so in four strains of A. gossypii.