Potential of the Bacillus thuringiensis toxin reservoir for the control of Lobesia botrana (Lepidoptera: Tortricidae), a major pest of grape plants

The potential of Bacillus thuringiensis Cry proteins to control the grape pest Lobesia botrana was explored by testing first-instar larvae with Cry proteins belonging to the Cry1, Cry2, and Cry9 groups selected for their documented activities against Lepidoptera. Cry9Ca, a toxin from B. thuringiensis, was the protein most toxic to L. botrana larvae, followed in decreasing order by Cry2Ab, Cry1Ab, Cry2Aa, and Cry1Ia7, with 50% lethal concentration values of 0.09, 0.1, 1.4, 3.2, and 8.5 microg/ml of diet, respectively. In contrast, Cry1Fa and Cry1JA were not active at the assayed concentration (100 microg/ml). In vitro binding and competition experiments showed that none of the toxins tested (Cry1Ia, Cry2Aa, Cry2Ab, and Cry9C) shared binding sites with Cry1Ab. We conclude that either Cry1Ia or Cry9C could be used in combination with Cry1Ab to control this pest, either as the active components of B. thuringiensis sprays or expressed together in transgenic plants.     

Generating susceptible strain and resistance status of field populations of Spodoptera exigua (Lepidoptera: Noctuidae) against some conventional and new chemistry insecticides in Pakistan

Two field populations of Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae) from Dera Ghazi Khan (D. G. Khan) and Multan, Pakistan, were tested for resistance to the 10 most commonly used insecticides in Pakistan by using a standard leaf disc bioassay on the F1 progeny. For comparison, a susceptible strain was generated from the Multan strain, which displayed lower LC50 values for most of the insecticides, by either mass rearing without exposure to insecticides or single-pair crosses against selected insecticides. The single-pair crosses generated a more susceptible strain than mass rearing. The D. G. Khan field strain was highly resistant to cypermethrin, profenofos, spinosad, abamectin, and chlorpyrifos and moderately resistant to deltamethrin, indoxacarb, and methoxyfenozide compared with susceptible lab strain. The Multan strain was highly resistant to profenofos and indoxacarb. Both field populations were susceptible to emamectin benzoate and lufenuron. Rotating these two insecticides with others that show very low, low, or moderate levels of resistance and have different modes of action may be useful for the effective management of this pest.     

Effects of lufenuron on Lobesia botrana (Lepidoptera: Tortricidae) egg, larval, and adult stages

The effect of the chitin synthesis inhibitor lufenuron was evaluated against different developmental stages of Lobesia botrana Den. & Schiff. (Lepidoptera: Tortricidae). Lufenuron fed to adults at 10 ppm reduced their fecundity and fertility, but it did not affect adult longevity. High activity was observed against L. botrana eggs with greater effect on 1-d-old eggs than on the other age classes and on eggs treated by direct contact rather than those laid on a previously treated surface. Eggs laid by treated adults showed the same effects during development as eggs treated by contact or those laid on a treated surface. Larvae that emerged from treated eggs could not perforate grape berries. Administered into the diet, lufenuron had a larvicidal effect, resulting in similar LC50 values for different instars: 0.07 ppm for first instars, 0.08 ppm for third instars, and 0.11 ppm for fifth instars. None of the larvae treated with sublethal concentrations throughout their life emerged as adults at the highest concentration (0.08 ppm), and only 70% emerged at the lowest concentration (0.0025 ppm).     

Resurgence of the cotton bollworm Helicoverpa armigera in northern Greece associated with insecticide resistance

Helicoverpa armigera has been controlled effectively with chemical insecticides in the major cotton crop production areas of northern Greece for many years. However, a resurgence of the pest was observed in 2010, which significantly affected crop production. During a 4‐year survey (2007 – 2010), we examined the insecticide resistance status of H. armigera populations from two major and representative cotton production areas in northern Greece against seven insecticides (chlorpyrifos, diazinon, methomyl, alpha‐cypermethrin, cypermethrin, gamma‐cyhalothrin and endosulfan). Full dose‐response bioassays on third instar larvae were performed by topical application. Lethal doses at 50% were estimated by probit analysis and resistance factors (RF) were calculated, compared to a susceptible laboratory reference strain. Resistance levels were relatively moderate until 2009, with resistance ratios below 10‐fold for organophosphates and carbamates and up to 16‐fold for the pyrethroid alpha‐cypermethrin. However, resistance rose to 46‐ and 81‐fold for chlorpyrifos and alpha‐cypermethrin, respectively in 2010, when the resurgence of the pest was observed. None of the known pyrethroid resistance mutations were found in the pyrethroid‐resistant insects. The possible association between resistance and H. armigera resurgence in Greece is discussed.     

Switching among natal and auxiliary hosts increases vulnerability of Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae) to insecticides

The role of insecticidal application and host plant resistance in managing Spodoptera exigua has been well documented, but the effect of different host plants, on which the pest cycles its population in the field, has seldom been investigated. Therefore, we have studied the vulnerability of S. exigua against commonly used insecticides (cypermethrin, chlorpyrifos, lufenuron, and emamectin benzoate) with different mode of actions when it switches its generations from natal to auxiliary hosts and vice versa. Different field populations being established on different host plants including castor, cauliflower, cotton, okra, and spinach were collected and reared in the laboratory before insecticidal bioassays. The role of larval diet and host plant switching on their response to tolerate applied insecticides was studied using leaf‐dip bioassay methods. Host switching demonstrated a significant role in altering the vulnerability of S. exigua populations to tested insecticides. Spodoptera exigua sourced from castor, when switched host to okra and spinach, exhibited 50% higher mortality when treated with emamectin benzoate. This trend in mortality was consistent upon complete host switch cycle (natal—auxiliary—natal host). However, the highest increase (92%) in vulnerability was recorded when the larvae were shifted to spinach from cotton. In general, chlorpyrifos and lufenuron had highest efficacies in terms of larval mortality. The findings of present studies provide insights to a better understanding the behavior of polyphagous pests and the role of different host plants in altering the susceptibility of these pests against applied insecticides. Ultimately the results warrant that due consideration should be given to cropping patterns and time of host switching by pest population during planning and executing chemical control.     

Evidence for p-glycoprotein modification of insecticide toxicity in mosquitoes of the Culex pipiens complex

Pesticide resistance has parallels with multi-drug resistance syndrome of tumours in clinical medicine, which has been linked to an ATP-dependent pump, p-glycoprotein (P-gp). P-gps pump drugs out of the cell, thereby reducing cellular concentrations of the chemical. P-gps have been found in several invertebrate species and have been shown to provide a defence against environmental xenobiotics, including pesticides. This study used a model cell culture system to investigate the interaction of pesticides with P-gp. Ivermectin and endosulfan were shown to be strong inhibitors of dye transport out of cells, which is a standard measure of P-gp modulation. We then investigated the action of a P-gp inhibitor, verapamil (calcium channel blocker), on insecticide toxicity to fourth-instar mosquito larvae of the Culex pipiens L. complex (Diptera: Culicidae). Verapamil increased toxicity to examples of three insecticide classes (cypermethrin, endosulfan, ivermectin), but not to chlorpyrifos (organophosphate). The discovery of a novel protective mechanism in mosquitoes, with a wide substrate range, has implications for the control of important pest and vector species.     

Influence of parasitism by Chelonus insularis Cresson (Hymenoptera: Braconidae) on the susceptibility of Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) to insecticides

The egg-larval parasitoid Chelonus insularis Cresson is a key parasitoid of the fall armyworm Spodoptera frugiperda (J. E. Smith) during the second field generation in the upper Magdalena River Basin, Colombia. In spite of selection pressure, the higher susceptibility of the second generation fall armyworm larvae to insecticides, compared with that of the first generation, suggests that the parasitism may be responsible for the apparent difference in susceptibility. Parasitized and non-parasitized 2nd-instar larvae of the fall armyworm were tested for susceptibility to chlorpyriphos, methomyl, cypermethrin, and Bacillus thuringiensis in the laboratory, using the leaf dip test. Parasitized larvae were up to 3.93 times more susceptible to chlorpyriphos, 3.71 times to methomyl, and 14.11 times to cypermethrin than non-parasitized larvae. The least effect of parasitism on susceptibility was found for B. thuringiensis. We discuss the negative influence of synthetic insecticide on the parasitoid population dynamics and its impact on insecticide resistance.     

Does natural larval parasitism of Lobesia botrana (Lepidoptera: Tortricidae) vary between years, generation, density of the host and vine cultivar?

Populations of European grapevine moth Lobesia botrana Denis & Schiffermüller and its larval parasitoids were studied for two consecutive years on an experimental insecticide-free vineyard in France planted with adjacent plots of five grape cultivars (Merlot, Cabernet Franc, Sauvignon, Cabernet Sauvignon and Sémillon) using a natural L. botrana population during the first year, and a natural population supplemented with artificially inoculated individuals during the second year. Levels of natural populations of larval parasitoids were measured by their parasitism rate. The ichneumonid Campoplex capitator Aubert was the most common species collected from L. botrana larvae. Its incidence was higher during the spring compared to summer. The overall parasitism rate found on the experimental vineyard varied from 23% in 2000 to 53% in 2001, and was mainly due to C. capitator. Parasitism was not affected by the grape cultivar on which the host developed but was positively correlated with the host density, per bunch or per stock, suggesting that among the five grape cultivars tested, C. capitator females probably do not discriminate between hosts feeding on different grape cultivars, but rather the densities of L. botrana larvae.     

Evaluation of resistance to different insecticides in field strains of Helicoverpa armigera (Lepidoptera: Noctuidae) in Punjab, Pakistan

Susceptibility of field populations of Helicoverpa armigera (Lepidoptera: Noctuidae) from Punjab, Pakistan to selected insecticides was evaluated using the leaf dip method. The resistance factors varied between populations. The general trends were low to moderate resistance to endosulfan; very low to low resistance to chlorpyrifos and profenofos; very low resistance to spinosad, indoxacarb and thiodicarb; moderate to high resistance to bifenthrin; high to very high resistance to cypermethrin; and very high resistance to lambda-cyhalothrin. Integrated pest management strategies aimed at reducing pesticide application, such as rotating the conventional insecticides having low resistance with newly developed synthetic insecticides and conserving natural enemies, are recommended.     

Mating disruption of Lobesia botrana (Lepidoptera: Tortricidae): effect of pheromone formulations and concentrations

The reluctance of Israeli vine growers to adopt the mating disruption technique to control the moth Lobesia botrana Den. & Schiff. has been attributed to the high cost of this method compared with that of traditional insecticide control. In this study, we tested the possibility of reducing the cost, first by testing different pheromone formulations (and thus open the market for competition) and second by reducing the pheromone concentration used in vineyards. Comparisons were made between two pheromone formulations--Shin-Etsu (Tokyo, Japan) at 165 g/ha and Concep (Sutera, Bend, OR) at 150 g/ha--and between two concentrations of Shin-Etsu, 165 and 110 g/ha. Pheromone dispensers were placed at the onset of the second moth generation. Comparison of the numbers of clusters infested with eggs and larvae of L. botrana showed no significant differences in the performance, either between the two formulations, or between the two tested concentrations. The results suggest that 1) the two formulations are equally effective, and 2) a low pheromone concentration is sufficient to maintain good control of small populations of L. botrana. However, when the population is high, pest control efficacy is not improved by increasing the pheromone concentration. Therefore, in the interest of reducing the relatively high cost of mating disruption, we emphasize that increasing the pheromone concentration does not provide improved control of high populations of L. botrana. The cost of mating disruption can be diminished by reducing the applied pheromone concentration and by using the least expensive pheromone formulations     

Comparison Between the Control Efficacy of Trichogramma evanescens Westwood (Hymenoptera: Trichogrammatidae) and Two Trichogramma cacoeciae Marchal Strains Against Grapevine Moth ( Lobesia botrana Den. & Schiff.), Depending on Their Release Density

In the first generation of the grape moth Lobesia botrana, Trichogramma evanescens spread to a greater extent than Trichogramma cacoeciae and parasitized most of the grape moth eggs. Reduction of damage to grapes was improved by doubling the number of T. evanescens release points in the first and second generations. In the second generation of the grape moth, T. cacoeciae was more efficient than T. evanescens . The highest proportion of individuals belonging to the reared and natural T. cacoeciae strains were found in the plot in which they were initially released. The total number of individuals emerging from parasitized eggs eggs was the same for each strain, which would suggest that neither predominated. Whichever T. cacoeciae strain considered, a density of 800 release points ha -1 did not induce greater parasitization than a density of 400 release points ha -1 .     

Could resistance to insecticides in Plutella xylostella (Lep., Plutellidae) be overcome by insecticide mixtures?

Abstract: To investigate if synergism occurs between pyrethroids, organophosphates and new insecticides, we tested representatives of these compounds (bifenthrin, chlorpyrifos, spinosad, indoxacarb and emamectin) against the diamondback moth (Plutella xylostella). Larvicidal activity of these insecticides was assessed separately and together on a susceptible strain (Lab‐UK) of P. xylostella as well as a field population collected from Multan. The field population showed significant resistance to chlorpyrifos (331 100‐fold), bifenthrin (45 200‐fold), emamectin (1800‐fold), spinosad (11‐fold) and indoxacarb (5600‐fold) when compared with the Lab‐UK population. When insecticides were mixed based on LC₅₀ and tested at serial concentrations against Lab‐UK, significant synergy (CI < 1) occurred between bifenthrin, spinosad and emamectin. In contrast, the interaction between bifenthrin and indoxacarb was additive (CI = 1). The toxicity of bifenthrin against the field population increased significantly (P < 0.01) when combined with spinosad, emamectin and indoxacarb. Synergistic effects could be attributed to the complementary modes of action by these insecticide classes acting on different components of nerve impulse transmission (which are not identical for bifenthrin and indoxacarb either). However, chlorpyrifos/bifenthrin mixture was not significantly different either from bifenthrin or chlorpyrifos alone, indicating an additive affect. In combination with spinosad and emamectin, tested against the resistant field population, the toxicity of chlorpyrifos increased significantly and even more so with indoxacarb. Mixtures could also give rise to multiple resistance that may extend across other chemical classes and thus become difficult to manage. Therefore, alternative strategies such as mosaics or rotations should be considered. That is, though synergistic effects have been found, this should not be followed up as a strategy to manage resistant field populations.     

Insecticide resistance in populations of the diamondback moth, Plutella xylostella (L.) (Lepidoptera: Plutellidae), from the state of Pernambuco, Brazil

The diamondback moth Plutella xylostella (L.) has a great economic importance in Brassicaceae crops in many parts of the world. Recurrent infestations of this pest in growing areas of Pernambuco state, Brazil, have led farmers to frequently spray their crops with insecticides. However, control failures by several insecticides have been alleged by farmers. The objective of this study was to check whether resistance to insecticides could explain these control failures in P. xylostella. Populations of P. xylostella from Pernambuco were collected between January and April 2009. The resistance ratios of P. xylostella populations were compared among five different active ingredients: abamectin, methomyl, lufenuron, indoxacarb, and diafenthiuron by leaf dipping bioassays using foliar discs of kale leaves. Mortality data were submitted to probit analysis. The P. xylostella populations showed variable response and significant resistance to one or more insecticides. The population from Bezerros County exhibited the highest resistance ratios to indoxacarb (25.3 times), abamectin (61.7 times), and lufenuron (705.2 times), when compared to the reference population. The populations from Bonito and Jupi Counties were 33.0 and 12.0 times more resistant to lufenuron and abamectin, respectively, when compared with the reference population. Resistance to methomyl was the least common, but not less important, in at least four populations. These results indicated that control failures were associated with resistance by some of the evaluated insecticides, reinforcing the need for resistance management in areas of the state of Pernambuco.     

Screening of the insecticidal activity of Bacillus thuringiensis strains against Lygus hesperus (Hemiptera: Miridae) nymphal population

Lygus hesperus Knight (Hemiptera: Miridae) is an economically important insect pest controlled primarily by chemical pesticides. Bacillus thuringiensis Berliner is a gram-positive bacterium that has been developed for the control of some insect pests in the orders Lepidoptera, Coleoptera, and Diptera. In this study, whole culture extracts of 94 B. thuringiensis strains from 83 serovars were added to an artificial diet and assayed against L. hesperus first and second instars. A total of five B. thuringiensis strains, B. thuringiensis variety thuringiensis, thuringiensis exotoxin +, morrisoni, tolworthi, and darmstadiensis generated > 98% mortality after 7 d of incubation. The screening was repeated with 117 alkali-solubilized trypsin-digested B. thuringiensis cultures and the same five B. thuringiensis strains showed nearly identical results. All five strains produce beta-exotoxin, which exhibits a wide host spectrum activity. No beta-exotoxin-minus B. thuringiensis strains showed significant toxicity against L. hesperus nymphs. The present work is one of the first thorough screenings of the wide diversity of the B. thuringiensis varieties for the control of L. hesperus nymphal populations.     

Evidence for resistance to pyrethroids and organophosphates in Plutella xylostella (Lepidoptera: Plutellidae) from Pakistan

The susceptibility of representative pyrethroid (cypermethrin, deltamethrin, lambdacyhalothrin, bifenthrin), organophosphate (chlorpyriphos, triazophos, profenophos) and new chemistry insecticides (spinosad, indoxacarb and emamectin) was investigated for 18 field populations of Plutella xylostella (Linnaeus) from three different zones in Pakistan. The LC(50) (mg ml(-1); 48 h) values of pyrethroids for various populations ranged from 0.19-1.88 for cypermethrin, 0.31-2.64 for deltamethrin, 0.08-1.16 for lambdacyhalothrin and 0.07-0.88 for bifenthrin. The LC(50) (mg ml(-1); 48 h) of organophosphates ranged from 0.52-5.67 for chlorpyriphos, 0.37-4.14 for triazophos and 0.03-2.65 for profenophos. The most probable reason for low toxicity of organophosphates and pyrethroids is the evolution of multiple resistance mechanisms; however, further studies are required to establish these mechanisms. When these same products were tested against a susceptible laboratory population (Lab-Pak), the new chemistry compounds were significantly more toxic than pyrethroids and organophosphates. The results are discussed in relation to integrated pest management and insecticide resistance management strategies for P. xylostella.     

Efficacy tests and determination of optimal spray timing values to control nantucket pine tip moth (Lepidoptera: Tortricidae) infestations

The Nantucket pine tip moth, Rhyaciona frustrana (Comstock), a common regeneration pest of loblolly pine, Pinus taeda L., has been shown to reduce tree volume yields through larval feeding. Chemical applications can be effective in protecting trees from the growth losses associated with this feeding, and optimum spray timing values are commonly used to reduce the number of necessary applications and to increase insecticide efficacy. Optimal spray timing values for the Georgia Piedmont were obtained for the following four insecticides available for use in loblolly pine plantations: permethrin (Pounce), lambda-cyhalothrin (Warrior T), spinosad (SpinTor 2 SC), and Bacillus thuringiensis variety kurstaki Berliner (Foray 48B). Optimal timing values were similar between the first and second generations for each of these compounds. All of the insecticides used in this study significantly reduced tip moth damage below the control treatment levels. Lambda-cyhalothrin was the most efficacious and had the longest spray timing window. B. thuringiensis was the least effective and had the shortest timing window. Spinosad and permethrin were similar in efficacy and spray timing values. This information is applicable to regions where there are three tip moth generations per year, as found in the southern Piedmont region and the coastal plain of Virginia and most of North Carolina.     

Insecticide susceptibility and detoxication enzyme activities among Coptotermes formosanus Shiraki workers sampled from different locations in New Orleans

Coptotermes formosanus Shiraki worker termites were sampled from 20 locations in the City Park area of New Orleans, LA. The termites were subsequently assayed to determine their susceptibility to cypermethrin, chlordane and chlorpyrifos, and detoxication enzyme activity. Cypermethrin was most toxic against Formosan subterranean termite workers, chlorpyrifos exhibited intermediate toxicity and chlordane was least toxic. A comparison of insecticide susceptibility between the most and least tolerant colonies revealed 1.9-, 1.7- and 1.8-fold differences in susceptibility for cypermethrin, chlorpyrifos and chlordane, respectively. As with the bioassay data, although significant differences were noted, a great deal of overlap was observed among the colonies for total cytochrome P450 content (difference of 2.2-fold between high and low value) aldrin epoxidation (3.6-fold) and cytosolic esterase (3.9-fold) activity. No significant differences were observed among the colonies for methoxyresorufin O-demethylase or glutathione S-transferase activity. Conversely, microsomal esterase activity varied greatly; a 38-fold difference was observed between the most (Cf1776) and least (Cf1387) active colonies. However, no significant correlation was observed between insecticide susceptibility and microsomal esterase activity. In fact, no significant correlations were observed between any of the enzyme activities measured and insecticide susceptibility. These results are discussed in the context of insecticide selection and future control effectiveness.     

Evaluating the efficacy of insecticides to control Sceliodes cordalis (Doubleday) (Lepidoptera: Crambidae) in eggplant

The efficacy of insecticides in controlling Sceliodes cordalis , eggfruit caterpillar, in eggplant was tested in four small plot trials because there has been a very limited range of insecticides available to manage this pest. Weekly applications of bifenthrin, flubendiamide, methoxyfenozide, chlorantraniliprole and spinosad and twice weekly applications of methomyl provided control as measured by a percentage of damaged fruit significantly lower than that in an untreated control. Twice weekly applications of methoxyfenozide, chlorantraniliprole or spinosad were not significantly more effective than weekly applications. Bacillus thuringiensis kurstaki , emamectin benzoate, indoxacarb, methomyl and pyridalyl applied weekly were ineffective, with percentages of damaged fruit not significantly different from the untreated control. These trials have identified a number of insecticides that could be used to manage S. cordalis , including several that would be compatible with integrated pest management programs in eggplant.     

Spatial analysis of Lobesia botrana (Lepidoptera: Tortricidae) male population in a mediterranean agricultural landscape in central Italy

The results obtained from the spatial analysis of pheromone-baited trap catch data of Lobesia botrana (Denis and Schiffermüller) males are reported. The research was undertaken in the Abruzzo region of central Italy. In the study area, vineyards (of Vitis vinifera L.) are the predominant cultivation, surrounded by hedgerows and small woodlots, and interspersed with cereal crops and olive groves. The main purpose of the study was to investigate the spatio-temporal dynamics of L. botrana, inside and outside vineyards, and to evaluate the effect of the landscape elements on pest distribution. A trend orientation over the experimental area was observed along the direction from northwest to southeast. Correlograms fitted using a spherical model showed in all cases an aggregated distribution and an estimated range having a mean of 174 m in 2005 and 116 m in 2006. Contour maps highlighted that spatial distribution of L. botrana was not limited to vineyards, but its presence is high particularly inside olive groves. The adult distribution on the experimental area changed during the season: hot spots of flight I were positioned inside olive groves; during flights II and III, they were concentrated in vineyards. L. botrana males were also captured in uncultivated fields, but never in high densities. Our results showed that a large proportion of the adult population of L. botrana inhabits areas outside those usually targeted by pest management programs. Thus, in Mediterranean agro-ecosystems, it is highly recommended to consider the whole landscape, with particular attention to olive crops.     

草地贪夜蛾幼虫对常用杀虫剂相对敏感基线的建立

【目的】本研究旨在建立草地贪夜蛾Spodoptera frugiperda对常用杀虫剂的相对敏感基线,为我国草地贪夜蛾的抗药性系统性监测提供依据。【方法】从玉米田采集草地贪夜蛾在室内不接触任何杀虫剂连续饲养5~7代,采用浸叶法和点滴法测定了草地贪夜蛾3龄幼虫以及采用饲料药膜法测定了草地贪夜蛾2龄幼虫对6大类共7种常用杀虫剂(甲维盐、乙基多杀菌素、虫螨腈、茚虫威、四氯虫酰胺、虱螨脲和氯虫苯甲酰胺)的敏感性,并据此建立了草地贪夜蛾幼虫对这7种杀虫剂的相对敏感基线。【结果】浸叶法测得上述7种药剂对草地贪夜蛾幼虫的LC₅₀值在0.054~12.131 mg/L之间;点滴法测得上述6种药剂(不含虱螨脲)对草地贪夜蛾幼虫的LD₅₀值在0.355~4.707 μg/g之间;饲料药膜法测得上述7种药剂对草地贪夜蛾幼虫的LC₅₀值在0.003~0.238 μg/cm²之间。【结论】采用3种生测方法建立了草地贪夜蛾幼虫对几种常用杀虫剂的相对敏感基线,为我国草地贪夜蛾的抗药性监测和化学防治提供了依据。