COMPARATIVE TOXICITY OF SIX INSECTICIDES ON THE RHINOCEROS BEETLE (COLEOPTERA: SCARABAEIDAE)

Headnote

Abstract

Strategus aloeus (Linnaeus, 1758) (Coleoptera: Scarabaeidae) is a dangerous pest of oil palms in the Americas, because the adults cause several kinds of damage and kill palm trees. Effective methods for pest management are needed urgently. Bioassays were conducted to compare the toxicity to S. aloeus of the insecticides: fipronil, imidacloprid, lambda- cyhalothrin, spinosad, thiacloprid and thiamethoxam. The toxicity of each insecticide to the adults of S. aloeus was determined as: (1) the LC50 and LC90 under laboratory conditions, after exposure of six concentrations of each insecticide applied in a semi-solid diet and used to feed each insect and (2) the mortality under semi-controlled field conditions after applications of insecticides into the beetle galleries in the oil palm tree. The mortality of S. aloeus was higher with fipronil, imidacloprid, lambda-cyhalothrin and thiamethoxam, while spinosad and thiacloprid were less effective. Higher mortalities were obtained with concentrations of 12.5, 25, 50 µL mL-1 for determining LC50 values and 50, 100 µL mL-1 for determining LC90 values during 72 h. The mortalities of S. aloeus had similar tendencies under laboratory and semi-controlled field conditions. Fipronil, imidacloprid, lambdacyhalothrin and thiamethoxam caused substantial mortality in S. aloeus and, thus, can be used rotationally in integrated pest management programs (IPM) against this pest in the oil palm plantations.

Key Words: Fipronil, imidacloprid, lambda-cyhalothrin, thiamethoxam, insect pest, neurotoxic insecticide, oil palm

resuMen

Strategus aloeus es una plaga peligrosa de la palma de aceite en América porque los adul- tos causan diversos daños y matan las palmas. Esto sugiere la necesidad de buscar mé- todos efectivos para el control de plagas. Se llevaron a cabo bioensayos para comparar la toxicidad del fipronil, imidacloprid, lambdacialotrina, spinosad, tiacloprid y tiametoxam sobre S. aloeus. La toxicidad de cada insecticida sobre adultos de S. aloeus fue determi- nada: (1) la concentración letal CL50 y CL90 en condiciones de laboratorio, después de la exposición de seis concentraciones de cada insecticida aplicadas en una dieta semisólida y utilizada para alimentar cada insecto y (2) mortalidad en condiciones semi-controladas de campo con la aplicación de insecticidas sobre galerías hechas por los insectos en árbo- les de palma de aceite. La mortalidad de S. aloeus fue mayor con fipronil, imidacloprid, lambdacialotrina y tiametoxam, mientras spinosad y tiacloprid fueron menos eficaces. Altas mortalidades se obtuvieron con concentraciones de 12.5, 25, 50 µL mL-1 para la CL50 y CL90, 100 µL mL-1 para la CL90 durante 72 horas. La mortalidad de S. aloeus fue similar en las condiciones controladas de laboratorio y de campo. La acción insecticida de esos compuestos puede ser producido debido a su capacidad de penetrar rápidamente en el cuerpo del insecto y afectar el sistema nervioso. Los insecticidas como fipronil, imida- cloprid, lambdacialotrina y tiametoxam causaron mortalidad en S. aloeus y, por lo tanto, pueden ser utilizados en programas de Manejo Integrado de Plagas (MIP) de esa plaga en plantaciones de palma de aceite.

Palabras Clave: Fipronil, imidacloprid, lambdacialotrina, tiametoxam, insecticidas neuro- tóxicos, insecto plaga, palma de aceite

The rhinoceros beetle, Strategus aloeus (Lin- naeus, 1758) (Coleoptera: Scarabaeidae) is a pest in oil palm (Elaeis guineensis Jacquin; Arecales: Arecaceae) plantations in the Americas. Strateg- us aloeus feeds on members of Arecaceae family such as Bactris gasipaes Kunth, Cocos nucifera (L.), Copernicia cerifera (Mart.), Elaeis oleifera (Kunth) and Phoenix dactylifera (L.) (Ratcliffe 1976; Genty et al. 1978; Chinchilla 2003). Strat- egus aloeus has a geographical distribution from the southern USA through Central America up to South America (Ratcliffe 1976; Genty et al. 1978). This beetle is nocturnal, and feeds and mates be- tween 18:30 and 05:30 h and 02:00 and 05:00 h, respectively (Pallares et al. 2000).

Adult S. aloeus colonize oil palm plantations in the replanting areas of 25-year-old oil palm stands. During replanting old palm trees are felled and replaced by young palms (Boyé & Au- bry 1973; Corley & Tinker 2003). The eggs of S. aloeus are laid in dead tissues of the palms; the larvae live in dead trunks or in the soil, and feed on decaying organic matter (Genty et al.1978; Ahumada et al. 1995). The adults attack 1-3 yr old palms by tunneling into the soil near the palm, and then boring their way into the plant just above the roots. Often, the growing point is reached and plant death occurs (Mariau 1994; Corley & Tinker 2003).

Various control methods such as application of entomopathogenic fungi and capture of adults by pheromone-baited traps have not been effective in protecting replanted crops where high populations of S. aloeus exist (Genty et al. 1978; Ahumada et al. 1995 Pallares et al. 2000). Other control proce- dures, such as collection of the immature stages on dead oil palms has a prohibitively high cost in commercial plantations (Bedford 1980; Corley & Tinker 2003). Insecticides such as carbamate and organophosphate compounds are the main method of control for S. aloeus (Genty et al. 1978; Ahuma- da et al. 1995). Carbamates and organophosphates act through contact and ingestion and cause neu- rotoxicity, which may be lethal (Bloomquist 1996; He 2000; Gilbert & Gill 2010). To protect oil palm crops, insecticides are applied inside the galleries of S. aloeus in the soil. Application of insecticides to the soil is an efficient method in managing the pest's populations by reducing palm tree damage and lessening the risk to human health and the environment (Darus & Basri 2000; Hartley 2002; Martínez 2013). Oil palm pests, such as Oryctes rhinoceros (L.) and Scapanes australis (Boisduval) (Coleoptera: Scarabaeidae), have been controlled successfully with insecticides applied in the soil galleries (Bedford 1976; Norman & Mohd Basri 1997; Corley & Tinker 2003).

Insecticides can be effective in protecting oil palm plantations, because they rapidly decimate dense insect pest populations (Mariau 1993; Nor- din et al. 2004). New insecticidal compounds that are achieving registration within the current regulatory environment of the US Environmen- tal Protection Agency (EPA) include chemical compounds that have reduced-risk and less im- mediate toxicity to the pest insects than the con- ventional insecticides (EPA 2011). These types of insecticides are potential new tools for use against the oil palm pest in the commercial plan- tations because of their suitability for inclusion in IPM programs.

This study compared the toxicity of 6 insecti- cides on S. aloeus adults under laboratory and field-controlled conditions, in order to contribute new control measures to IPM programs against S. aloeus in oil palm plantations.

MateriaLs and Methods

Insects

In the field, 2,683 pupae of S. aloeus (^ = 1314, ^ = 1369) were hand collected from dead palm stems in 2 yr-old commercial plantations of the oil palm in the municipality of Puerto Wilches, Santander, Colombia (N 07° 20' -W 73° 54'), with average conditions of 28.46 °C, 75-92% RH, 1,450-2,250 h sunshine/year and 2,168 mm an- nual rainfall. The insects were placed in metallic boxes (70 × 70 × 80 cm) covered with a nylon mesh and transferred to the Entomology Laboratory of the Brisas Oil Palm Plantation, Puerto Wilches, Santander, Colombia. Pupae of S. aloeus were iso- lated in plastic containers (15 × 10 cm) containing soil and a moistened cotton ball. The insects were maintained at 28 ± 1 °C, 75 ± 5% RH and 12:12 h L:D photoperiod until adult emergence.

Adults were placed in glass containers (30 × 30 × 30 cm) covered with a nylon mesh with a 15 cm deep soil layer maintained at 80% RH by periodic watering rather by a sprinkler. The beetles were fed on semi-solid diet daily (10 g of stem palm mer- istem + 10 g of sugarcane + 10 g of banana + water in a 5:5:2:1 ratio) (Pallares et al. 2000). The diet ingredients were cut, chopped and mixed in a labo- ratory blender (Waring LBC15; 3,500-20,000 rpm). The semi-solid diet was sterilized with ultraviolet light for 30 min and stored at -5 °C. Photophase and scotophase periods were simulated with white and red lights (IRO 110V 60W; Toshiba Lighting and Technology Corp., Tokyo, Japan), respectively. Healthy S. aloeus adults without amputations or malformations were used in the bioassays.

Insecticides

Six different neurotoxic insecticides were used in this study. The following commercial insecti- cides were tested at their maximum label rates: fipronil (Clap® SC, Bayer, Germany), 200 g L-1; imidacloprid (Confidor® SL, Cyanamid, Bel- gium), 200 g L-1; lambda-cyhalothrin (Karate® EC, Zeneca, Wilmington, USA), 25 g L-1; spinosad (Tracer® SC, Dow Agrosciences, USA), 120 g L-1; Thiacloprid (Calypso® SC, Cyanamid, Belgium), 480 g L-1; and thiamethoxam (Actara® SC, Syn- genta, Canada), 240 g L-1. These insecticides were diluted in 1 L water to produce a stock solution by adjusting 100 g L-1 per insecticide and to obtain the required concentrations.

Insecticide Bioassay

Insecticide efficacy was determined by calculat- ing the lethal concentrations (LC50 and-LC90) and lethal time (LT50 and LT99) values under laboratory conditions for each formulation. Six concentrations of each insecticide besides the control (distilled water), were adjusted in 1 mL stock solution (in- secticide and distilled water): 3.12, 6.25, 12.5, 25, 50 µL and 100 µL. For each insecticide, aliquots were taken from the stock solution and mixed with distilled water in 5 mL glass vials. Different con- centrations of the insecticides were applied in 1 µL solution onto 5 g of semi-solid diet and used to feed each insect. Fifty insects were used per con- centration and were placed individually in plastic containers (15 × 10 cm) with a perforated lid and maintained in the dark. The number of dead in- sects in each cage was counted after insecticide exposure at intervals of 2 h over 6 days.

Assays for Mortality under Field Conditions

One hundred, 6-month-old oil palms ('Tenera' cv, 'Deli' × 'Ghana') and 100 adults of S. aloeus were used for each insecticide in the controlled field test. These palms were planted into soil in polyethylene bags and maintained in the greenhouse at a tem- perature of 27 ± 2 °C, 75-80% RH and 25% shadow. One insect was caged in each oil palm a nylon trap (0.5 × 0.5 × 1.20 m). Treatments consisted of adding each insecticide at the calculated LC90 concentration and distilled water as the control with 4 replications per treatment. The LC90 concentration of each in- secticide was applied directly on the galleries while distilled water was applied to the controls. Applica- tions of 500 mL per gallery were made by a manual pump (Royal Condor, 10 L capacity). The applica- tion was evaluated after 5 days, when the insects build their galleries. Mortality of S. aloeus caused by insecticides was recorded every day during 15 days, and was corrected both in the laboratory and field bioassays according to Abbott (1925)

Statistics

The LC , LT and their confidence limits were determined by logistic regression based on the concentration probit-mortality (Finney 1964), with the program XLSTAT-PRO v.7.5 for Win- dows (XLSTAT 2004). Mortality data from the controlled field test were analyzed by one-way ANOVA. Mortality variables were summarized in percentages and the data were transformed by arcsine square root. Tukey's Honestly Significant Difference test (HSD) was used for comparison of the means at the 5% significance level (PROC ANOVA) (SAS 2002).

resuLts

Insecticide Susceptibility of Strategus aloeus

The highest mortalities were obtained with 50 and 100 µL mL-1 of the above mentioned insecti- cides. The 3 different lethal concentrations levels (LC50, LC90 and LC99) (Table 1) of each insecticide was estimated by Probit (χ2; P < 0.001). The LC50- 90 values indicated that fipronil (χ2 = 37.81; df = 5), lambda-cyhalothrin (χ2 = 42.52; df = 5) and thiamethoxam (χ2 = 48.43; df = 5) were the most toxic compounds to the S. aloeus adults followed by thiacloprid (χ2 = 27.84; df = 5). (LC50-90) value was lower with imidacloprid (χ2 = 13.07; df = 5) and spinosad (χ2 = 12.02; df = 5). Mortality was always < 1% in the control.

LT50 values were recorded when the pest was exposed from 21 to 144 h (1-6 days) to the various insecticides each applied at 6.25, 12.5, 25, 50 and 100 µL mL-1 (Fig. 1). The elapsed times for LC50 values were recorded from 24 h to 144 h for the insecticide concentrations of 6.25 and 100 µL mL-1 The LT50 values recorded at the higher and lower concentrations were between 21.81 and 111.9 h for fipronil, 25.53 and 85.71 h for imidacloprid, 48.97 and 120.1 h for lambda-cyhalothrin, 72.01 and 144.1 h for thiamethoxam, 77.41 and 105.8 h for spinosad, and to 100.1 and 120.1 h for thiaclo- prid. The elapsed times for the LC99 values were recorded from 120 to 144 h for the insecticide concentrations of to 50 and 100 µL mL-1. The LT99 values recorded at the higher and lower concen- trations were 97.96 h to fipronil, 125.1 h to imida- cloprid, 118.8 h to lambda-cyhalothrin and 142 h to thiamethoxam. It was not possible to estimate LT99 values for spinosad and thiacloprid at any of the exposure periods used.

Mortality in Controlled Field Tests

The mortality effects caused by the tested in- secticides on the S. aloeus adults were statistically different in controlled field conditions using the previously estimated concentrations for the LC90 values (F1,23 = 7.85; P < 0.05). Fipronil, lambda- cyhalothrin and thiamethoxam caused mortalities of 84.1 ± 9.5%, 88.3 ± 3.7% and 89.9 ± 12.4%, re- spectively, followed by mortalities by thiacloprid, imidacloprid and spinosad of 78.9 ± 8.9%, 73.9 ± 7.9% and 67.3 ± 7.3%, respectively Mortality never exceeded 3.14 ± 0.3% in the control (Fig. 2).

disCussion

The toxicity profiles for the 6 insecticides against the rhinoceros beetle, S. aloeus were de- termined from the bioassays in the laboratory and controlled field conditions. The insecticides fipronil, imidacloprid, lambda-cyhalothrin, spi- nosad, thiacloprid and thiamethoxam caused substantial mortality of the S. aloeus adults un- der laboratory conditions. The best results were obtained with concentrations of 50 and 100 µL mL-1. The susceptibility of the Scarabaeids may vary with exposure in the different concentra- tions of insecticides (Villani et al. 1988; Cowles & Villani 1996; Gilbert & Gill 2010). Different stud- ies show that Macrodactylus subspinosus (Fabri- cius), Oryctes rhinoceros (L.), and Popillia japon- ica (Newman) were susceptible to the insecticide concentration ranges applied in the food (Isaacs et al. 2004; Baumler & Potter 2007; Sreeletha & Geetha 2012).

The insects exposed to the neurotoxic insec- ticides: fipronil, thiacloprid and thiamethoxam displayed altered locomotion activity, and muscle contractions that were observed at high concen- trations in LC50-90 test. In contrast, the adults of S. aloeus gradually lost mobility when they were exposed to the lambda-cyhalothrin. In some indi- viduals, the paralysis was constant with concen- trations near the LC50 without recovery signs. The mode of action of the fipronil involves blocking the γ-aminobutyric acid (GABA)-gated chloride chan- nel which is also the target for cyclodiene insec- ticides such as endosulfan and dieldrin (Cole et al. 1993; Hainzl & Casida 1996; Durham et al. 2002). Lambda-cyhalothrin and other pyrethroids are known to cause rapid paralysis in inverte- brates by disrupting nerve conduction; their pri- mary site of action are the voltage-gated sodium channels (Bloomquist & Miller 1986; Baumler & Potter 2007). Imidacloprid, thiacloprid and thia- methoxam appear to interfere with the nicotinic acetylcholine receptors located in the post-syn- aptic membrane, disrupting normal nerve func- tion (Grewal et al. 2001; Matsuda 2001; Tan et al. 2008). Spinosad is a neurotoxin that targets the nicotinic acetylcholine receptor and the GA- BA receptors, and causes cessation of feeding fol- lowed later by paralysis and death (Salgado 1998; Crouse et al. 2001). In this study, the LC50-90 val- ues indicated that lethality of imidacloprid and spinosad were lower on S. aloeus using the evalu- ated concentrations. However, the lethalities of fipronil, lambda-cyhalothrin, thiacloprid and thiamethoxam on S. aloeus may have advantages by their mode of action on this insect and could be applied in rotation, under field conditions.

The insecticides evaluated required extended periods of time to achieve the mortality of S. al- oeus. The LT50 varied from 21 to 144 h, whereas the LT90 ranged from 120 and 144 h. The lethal ef- fect of fipronil was observed quickly (in 21 h), fol- lowed by imidacloprid (25 h), lambda-cyhalothrin (48 h), thiamethoxam (72 h), while spinosad and thiacloprid showed lethal effects at 76 h and 100 h, respectively. In this study, the comparative ef- fects on S. aloeus between the 6 insecticides were observed at various time points. Immediate lethal effect of insecticides are not essential to protect crops, unless the target insects vector viruses or other dangerous pathogens. Some insecticides cause cessation of feeding long before the target insects dies. However, a quick-acting insecticide is generally considered essential for impregna- tion of the palm stems; in this case, the beetles were able to feed, even if they died later. Neuro- toxic insecticides can reduce injuries of commer- cial crops caused by the nocturnal beetles (Drink- water 2003; Isaacs et al. 2004).

In the controlled field tests, the lethal effects of the insecticides on S. aloeus were consistent with those observed under laboratory condi- tions. Fipronil, lambda-cyhalothrin, thiacloprid and thiamethoxam showed strong lethal effects on S. aloeus, whereas imidacloprid and spinosad were less toxic. However, when the insecticides were applied to the insect in the field, the mortal- ity levels were lower than those obtained under laboratory conditions. One possible reason for the efficacy of the insecticides under the field condi- tions could be contact exposure on the body. In this bioassay, it is difficult to accurately know the amount of the insecticide absorbed by the insect, but the lethal effects caused by these insecticides on S. aloeus showed continuity of that trend with the application of the LC90 concentration. The le- thal effects of the insecticides and their effective- ness have also been studied in other pests under field conditions with fipronil being a potent con- trol agent of Diloboderus abderus Sturm larvae, lambda-cyhalothrin and thiacloprid being a po- tent in controlling P. japonica larvae and adults, and thiamethoxam being a potent against Het- eronychus arator Fabricius (Silva & Boss 2002; Drinkwater 2003; Baumler & Potter 2007).

This study showed the potential of 6 insec- ticides to control the S. aloeus adults in oil palm plantations. The toxicity of these insec- ticides with different modes of action may en- able the control of the S. aloeus populations and may reduce injuries caused by this insect on E. guineensis. Fipronil, lambda-cyhalothrin, thia- cloprid and thiamethoxam have lethal effects on S. aloeus with the potential to manage its field populations. In order to prevent or retard the de- velopment of insecticide resistance, insecticides with different modes of action should be used in rotation as indicated by Insecticide Resistance Action Committee (IRAC 2014). The IRAC mode of action groups to which the above insecticides belong are as follows: fipronil (2B, GABA-gated chloride channel antagonists), lambda-cyhalo- thrin (3A, Sodium channel modulators), imi- dacloprid, thiacloprid and thiamethoxam (4A, Nicotinic acetylcholine receptor [nAChR] ago- nists), and spinosad (5, Nicotinic acetylcholine receptor [nAChR] allosteric activators). Thus these insecticides belong to four IRAC mode of action groups, and their field applications should be rotated accordingly.

aCknowLedgMents

We thank Angel Contreras for technique support. To Oleagionas Las Brisas (Colombia), Conselho Nacional de Desenvolvimento Científico e Tecnológico CNPq (Brasil), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior CAPES (Brasil), and Fundação de Amparo a Pesquisa do Estado de Minas Gerais FAPEMIG (Brasil).

References

referenCes Cited

abbott, w. s. 1925. A method of computing the effective- ness of an insecticide. J. Econ. Entomol. 18: 265-266.

ahuMada, M. L., CaLvaChe, h., Cruz, M. a., and Luque, J. e. 1995. Strategus aloeus (L.) (Coleoptera: Scarabaeidae): biología y comportamiento en Puerto Wilches (Santander). Palmas 16: 9-16.

bauMLer, r. e., and Potter, d. a. 2007. Knockdown, residual, and antifeedant activity of pyrethroids and home landscape bioinsecticides against Japanese beetles (Coleoptera: Scarabaeidae) on linden foliage. J. Econ. Entomol. 100: 451-458.

bedford, g. o. 1976. Observations on the biology and ecology of Oryctes rhinoceros and Scapanes australis (Coleoptera: Scarabaeidae: Dynastinae): Pests of co- conut palms in Melanesia. Australian J. Entomol. 15: 241-251.

bedford, g. o. 1980. Biology, ecology and control of palm rhinoceros beetles. Annu. Rev. Entomol. 25: 309-339.

bLooMquist, J. r. 1996. Ion channels as targets for in- secticides. Annu. Rev. Entomol. 41: 163-190.

bLooMquist, J. r., and MiLLer, t. a. 1986. Sodium channel neurotoxins as probes of the knockdown re- sistance mechanism. Neurotoxicology 7: 217-224.

boye, P., and aubry, M. 1973. Replantation des palm- eraies industrielles. Methode de preparation de ter- rain et de protection contre l'Oryctes en Afrique de l'Ouest. Oléagineux 28: 175-176.

ChinChiLLa, C. M. 2003. Manejo integrado de problemas fitosanitarios en palma aceitera Elaeis guineensis en América Central. Manejo Integrado de Plagas y Agro- ecología 67: 69-82.

CoLe, L. M., niChoLson, r. a., and Casida, J. e. 1993. Action of phenylpyrazole insecticides at the GABA- gated chloride channel. Pestic. Biochem. Physiol. 46: 47-54.

CorLey, r. h. v., and tinker, P. b. 2003. The oil palm, 4th. Blackwell Science Ltd, USA.

CowLes, r. s., and viLLani, M. g. 1996. Susceptibility of Japanese beetle, oriental beetle, and European cha- fer (Coleoptera: Scarabaeidae) to halofenozide, and insect growth. J. Econ. Entomol. 89: 1556-1565.

Crouse, g. d., sParks, t. C., sChoonover, J., gifford, J., driPPs, J., bruCe, t., Larson, L. L., garLiCh, J., hatton, C., hiLL, r. L., worden, t. v., and Mar- tynow, J. g. 2001. Recent advances in the chemistry of spinosyns. Pest Mgt. Sci. 57: 177-185.

darus, a., and basri, M. w. 2000. Intensive IPM for management of oil palm pests. Oil Palm Bull. 41: 1-14.

drinkwater, t. w. 2003. Bioassays to compare the sys- temic activity of three neonicotinoids for control of Heteronychus arator Fabricius (Coleoptera: Scara- baeidae) in maize. Crop Prot. 22: 989-993.

durhaM, e. w., siegfried, b. d., and sCharf, M. e. 2002. In vivo and in vitro metabolism of fipronil by larvae of the European corn borer Ostrinia nubilalis. Pest Mgt. Sci. 58: 799-804.

environMentaL ProteCtion agenCy 2011. EPA's Pes- ticides industry sales and usage, 2006 and 2007 Mar- ket estimates. <http://www.epa.gov/oppbead1/pest- sales/. Accessed I-2012>.

finney, d. J. 1964. Probit analysis: a statistical treat- ment of the sigmoid response curve. Cambridge Uni- versity Press, London.

genty, P., desMier de Chenon, d., and Morin, J. 1978. Ravageurs du palmier á huile en Amérique Latine. Oléagineux 33: 325-419.

giLbert, L. i., and giLL, s. s. 2010. Insect control bio- logical and synthetic agents. Academic Press-Elsevier B.V. London, UK.

grewaL, P. s., Power, k. t., and shetLar, d. J. 2001. Neonicotinoid insecticides alter diapause behavior and survival of overwintering white grubs (Coleop- tera: Scarabaeidae). Pest Mgt. Sci. 57: 852-857.

hainzL, d., and Casida, J. e. 1996. Fipronil insecticide: Novel photochemical desulfinylation with retention of neurotoxicity (insecticide action and environmen- tal persistence). Proc. Natl. Acad. Sci. USA 93: 12764- 12767.

hartLey, M. J. 2002. Rationale and methods for conserv- ing biodiversity in plantation forests. Forest Ecol. Mgt. 155: 81-95.

he, f. 2000. Neurotoxic effects of insecticides-current and future research: a review. Neurotoxicology 21: 829-35.

iraC. 2014. IRAC MoA Classification Scheme, Version 7.3. http://www.irac-online.org/teams/mode-of-action/ .Accessed V-24-2014.

isaaCs, r., MerCader, r. J., and wise, J. C. 2004. Ac- tivity of conventional and reduced-risk insecticides for protection of grapevines against the rose chafer, Macrodactylus subspinosus (Coleoptera: Scarabaei- dae). J. Appl. Entomol. 128: 371-376.

Mariau, d. 1993. Insecticides recommended against oil palm and coconut pests. Oléagineux 48: 530-532.

Mariau, d. 1994. Phytosanitary monitoring of oil palm and coconut plantations. Oléagineux 49: 250-254.

Martínez, o. L., PLata-rueda, a., and Martínez, L. C. 2013. Oil palm plantations as an agroecosystem: impact on integrated pest management and pesti- cide use. Outlooks Pest Mgt. 24: 225-229.

Matsuda, k., buCkinghaM, s. d., kLeier, d., rauh, J. J., grauso, M., and satteLLe, d. b. 2001. Neo- nicotinoids: insecticides acting on insect nicotinic acetylcholine receptors. Trends Pharmacol. Sci. 22: 573-580.

norMan, h. k., and Mohd basri, w. 1997. Status of rhinoceros beetle, Oryctes rhinoceros (Coleoptera: Scarabaeidae) as a pest of young oil palm in Malay- sia. The Planter 73: 5-21.

nordin, a. b. a., siMeh, M. a., aMiruddin, M. n., weng, C. k., and saLaM, b. a. 2004. Economic fea- sibility of organic palm oil production in Malaysia. Oil Palm Ind. Econ. J. 4: 29-38.

PaLLares, C. h., aLdana, J. a., CaLvaChe, h., raMírez, P., r°Chat, d., Luque, e., and Correa, n. 2000. Análisis del comportamiento y comuni- cación química intraespecífica en Strategus aloeus (L.) (Coleoptera, Scarabaeidae - Dynastinae). Pal- mas 21: 185-194.

ratCLiffe, b. C. 1976. A revision of the genus Strat- egus (Coleoptera: Scarabaeidae). Univ. Nebraska Museum. Papers in Entomol. 144: 1-115. http://digi- talcommons.unl.edu/entomologypapers/144

saLgado, v. L. 1998. Studies on the mode of action of spinosad: insect symptoms and physiological corre- lates. Pestic. Biochem. Physiol. 60: 91-102.

siLva, M. t. b., and boss, a. 2002. Controle de larvas de Diloboderus abderus com inseticidas em trigo. Cienc. Rural 32: 191-195.

sreeLetha, C., and geetha, P. r. 2012. Pesticidal ef- fects of Annona squamosa L. on male Oryctes rhinoc- eros Linn. (Coleoptera: Scarabaeidae) in relation to reproduction. Curr. Biotic. 6: 8-21.

sas institute. 2002. The SAS System for Windows, release 9.0. SAS Institute, Cary, N.C.

tan, J., saLgado, v.L., and hoLLingworth, r. M. 2008. Neural actions of imidacloprid and their in- volvement in resistance in the Colorado potato bee- tle, Leptinotarsa decemlineata (Say). Pest Mgt. Sci. 64: 37-47.

viLLani, M. g., wright, r. J., and baker, P. b. 1988. Differential susceptibility of Japanese beetle, orien- tal beetle, and European chafer (Coleoptera: Scara- baeidae) larvae to five soil insecticides. J. Econ. En- tomol. 81: 785-788.

xLstat 2004. XLSTAT for Excel. Addinsoft, UK.