Risk and Toxicity Assessment of a Potential Natural Insecticide, Methyl Benzoate, in Honey Bees (Apis mellifera L.).

Methyl benzoate (MB) is a component of bee semiochemicals. Recent discovery of insecticidal activity of MB against insect pests provides a potential alternative to chemical insecticides. The aim of this study was to examine any potential adverse impact of MB on honey bees. By using two different methods, a spray for contact and feeding for oral toxicity, LC50s were 236.61 and 824.99 g a.i./L, respectively. The spray toxicity was 2002-fold and 173,163-fold lower than that of imidacloprid and abamectin. Piperonyl butoxide (PBO, inhibiting P450 oxidases [P450]) significantly synergized MB toxicity in honey bees, indicating P450s are the major MB-detoxification enzymes for bees. Assessing additive/synergistic interactions indicated that MB synergistically or additively aggravated the toxicity of all four insecticides (representing four different classes) in honey bees. Another adverse effect of MB in honey bees was the significant decrease of orientation and flight ability by approximately 53%. Other influences of MB included minor decrease of sucrose consumption, minor increase of P450 enzymatic activity, and little to no effect on esterase and glutathione S-transferase (GST) activities. By providing data from multiple experiments, we have substantially better understanding how important the P450s are in detoxifying MB in honey bees. MB could adversely affect feeding and flight in honey bees, and may interact with many conventional insecticides to aggravate toxicity to bees. However, MB is a relatively safe chemical to bees. Proper formulation and optimizing proportion of MB in mixtures may be achievable to enhance efficacy against pests and minimize adverse impact of MB on honey bees.

Microarray analysis of gene regulations and potential association with acephate-resistance and fitness cost in Lygus lineolaris.

The tarnished plant bug has become increasingly resistant to organophosphates in recent years. To better understand acephate resistance mechanisms, biological, biochemical, and molecular experiments were systematically conducted with susceptible (LLS) and acephate-selected (LLR) strains. Selection of a field population with acephate significantly increased resistance ratio to 5.9-fold, coupled with a significant increase of esterase activities by 2-fold. Microarray analysis of 6,688 genes revealed 329 up- and 333 down-regulated (≥2-fold) genes in LLR. Six esterase, three P450, and one glutathione S-transferase genes were significantly up-regulated, and no such genes were down-regulated in LLR. All vitellogenin and eggshell protein genes were significantly down-regulated in LLR. Thirteen protease genes were significantly down-regulated and only 3 were up-regulated in LLR. More than twice the number of catalysis genes and more than 3.6-fold of metabolic genes were up-regulated, respectively, as compared to those down-regulated with the same molecular and biological functions. The large portion of metabolic or catalysis genes with significant up-regulations indicated a substantial increase of metabolic detoxification in LLR. Significant increase of acephate resistance, increases of esterase activities and gene expressions, and variable esterase sequences between LLS and LLR consistently demonstrated a major esterase-mediated resistance in LLR, which was functionally provable by abolishing the resistance with esterase inhibitors. In addition, significant elevation of P450 gene expression and reduced susceptibility to imidacloprid in LLR indicated a concurrent resistance risk that may impact other classes of insecticides. This study demonstrated the first association of down-regulation of reproductive- and digestive-related genes with resistance to conventional insecticides, suggesting potential fitness costs associated with resistance development. This study shed new light on the understanding of the molecular basis of insecticide resistance, and the information is highly valuable for development of chemical control guidelines and tactics to minimize resistance and cross-resistance risks.

Inheritance, fitness cost and mechanism of resistance to tebufenozide in Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae).

BACKGROUND: Tebufenozide has been used as a key insecticide for controlling beet armyworm, Spodoptera exigua (Hübner). To evaluate the risk of resistance evolution and to develop a better resistance management strategy, a field-collected population was selected with tebufenozide in the laboratory. Thereafter, the inheritance and fitness cost of tebufenozide resistance in S. exigua were investigated. RESULTS: After being selected with tebufenozide for 61 generations, S. exigua developed a 92-fold resistance to the chemical. The degrees of dominance for the reciprocal cross progeny were - 0.2698 and - 0.2785. The resistant strain had a relative fitness of 0.71, with substantially lower rates of larval survival, pupal weight, pupation and oviposition per female, and prolonged larval and pupal duration. PBO significantly increased the toxicity of tebufenozide (SR = 2) against resistant insects, and DEM and DEF also showed synergism with tebufenozide. CONCLUSION: Tebufenozide resistance in S. exigua was inherited as autosomal, incompletely recessive and controlled by more than one gene. Development of the resistance may cost significant fitness for the resistant population. Mixed-function oxidases might play an important role in tebufenozide resistance in S. exigua. This study provided valuable information for further understanding tebufenozide resistance and for facilitating the development of resistance management strategies.

Imidacloprid susceptibility survey and selection risk assessment in field populations of Nilaparvata lugens (Homoptera: Delphacidae).

Imidacloprid has been used for many years to control planthopper Nilaparvata lugens (Stål) (Homoptera: Delphacidae) in China. To provide resistance assessment for the national insecticide resistance management program, we collected a total of 42 samples of the planthoppers from 27 locations covering eight provinces to monitor their dose responses and susceptibility changes to imidacloprid over an 11-yr period (1996-2006). Results showed that most field populations maintained susceptibility from 1996 to 2003 except for a population from Guilin, Guangxi, in 1997, which showed a low level of resistance to imidacloprid. However, surveys conducted in 2005 indicated that 16 populations from six provinces quickly developed resistance with resistance ratios ranging from 79 to 811. The data collected in 2006 revealed that the resistance levels in 12 populations collected from seven different provinces decreased slightly (RR = 107-316), except the Tongzhou population (Jiangsu Province), which developed 625-fold resistance. Dominant and intensive use of imidacloprid in a wide range of rice, Oryza savita L., growing areas might be a driving force for the resistance development. Migration of the insect also significantly boosted the resistance levels due to extensive and intensive use of imidacloprid in emigrating areas and continuous postmigration sprays of the chemical. In addition, laboratory resistance selection using imidacloprid showed that resistance ratio increased to 14-fold after 27 generations, suggesting that quick resistance development might be associated with more frequent applications of the insecticide in recent years.