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Rosala Servn, Ricardo Aguilar, Jos L. Martnez, Enrique Troyo, Alfredo Ortega Monitoring of resistance to three insecticides on pepper weevil (anthonomus eugenii) in populations from Baja California Sur, Mexico Interciencia, vol. 27, nm. 12, diciembre, 2002, pp. 691-694, Asociacin Interciencia Venezuela
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Interciencia, ISSN (Printed Version): 0378-1844 interciencia@ivic.ve Asociacin Interciencia Venezuela

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MONITORING OF RESISTANCE TO THREE INSECTICIDES ON PEPPER WEEVIL (Anthonomus eugenii) IN POPULATIONS FROM BAJA CALIFORNIA SUR, MEXICO
Rosala Servn, Ricardo Aguilar, Jos L. Martnez, Enrique Troyo and Alfredo Ortega

SUMMARY Toxicity levels of three insecticides, carbaryl, endosulfan and methomyl, were determined in populations of the pepper weevil Anthonomus eugenii. Insect populations were obtained from three locations in Baja California Sur, Mexico: Benito Jurez (2753N, 11346W), San Jos Viejo (2307N, 10943W), and San Juan de los Planes (2358N, 10956W). The highest LC50 values were found for carbaryl (178.4, 635.4 and 5462.1 g/ml), endosulfan was intermediate (36.5, 34.1 and 47.1 g/ml, respectively); and the lowest LC50 values corresponded to methomyl (1.3, 1.7 and 2.7 RESUMEN Se determinaron los niveles de toxicidad en poblaciones de picudo del chile Anthonomus eugenii a tres insecticidas, carbaril, endosulfan y metomil. Las poblaciones de insectos fueron obtenidas de tres localidades en Baja California Sur, Mxico: Benito Jurez (2753N 11346W), San Jos Viejo (2307N, 10943W) and San Juan de los Planes (2358N, 10956W). Las CL50 ms elevadas corresponden al carbaril (178,4;635,4 y 5462,1 gr/ml); endosulfan tuvo valores intermedios (36,5; 34,1 y 47,1 g/ml); mientras que los valores ms bajos correspondieron a metomil (1,3; 1,7 y 2,7 g/ml), para Benito Jurez, San Jos Viejo y San Juan de los Planes respectivamente. Se puede concluir que las poblaciones de picudo del chile de San Juan de los Planes son resistentes a los tres insecticidas utilizados, teniendo mayor resistencia a carbaril, en comparacin con las poblaciones provenientes de San Jos el Viejo y Benito Jurez. Los niveles de susceptibilidad corresponden con las cantidades de insecticidas usados en contra del picudo del chile en las tres localidades de estudio y son una respuesta a la presin selectiva ejercida por esos compuestos. g/ml), to Benito Jurez, San Jos Viejo and San Juan de los Planes, respectively. It is concluded can be inferred that pepper weevil populations in the farming zone of San Juan de los Planes are resistant to the three insecticides used, and are significantly more resistant to carbaryl, than are pepper weevil populations in San Jos Viejo and Benito Jurez. Levels of insecticide susceptibility correspond to the amounts of insecticides used against the pepper weevil in the three locations, and are a response to the selective pressure exerted by those chemicals.

Introduction The pepper weevil, Anthonomus eugenii Cano (Coleoptera: Curculionidae) is the primary insect pest of the different varieties of chili pepper grown in Mexico (Bujanos, 1993). This insect causes premature maturation, evidenced by the fall of floral buds and fruit, and by a darkening or

yellowing of the fruit. The damage is caused by the larvae, which feed on the pulp and seeds of developing fruits. Flowers are susceptible to damage from ovoposition and from feeding by adult weevils, thus causing a significant reduction in the fruit production per plant (Pacheco-Covarrubias, 1985; Bernot, 1992). In Mexico, economic loss caused

by the pepper weevil may range from 50 to 100% of total pepper production (Laborde and Pozo, 1982; Bujanos, 1993; Riley and King, 1994; SAGAR, 1997). To solve this problem, several types of insecticide have been used as chemical control methods, especially carbaryl, endosulfan and methomyl. Avila (1987) notes that to obtain good

crops, farmers have invested up to 45% of the total crop cost in insecticides recommended for the chemical control of the pepper weevil. In response to increasing pesticide application, the pepper weevil has developed efficient resistance mechanisms against these chemicals. In the state of Baja California Sur, despite arid condi-

KEYWORDS / Anthonomus eugenii / Carbaryl / Endosulfan / Insecticides / Methomyl / Pepper Weevil /


Received: 06/13/2002. Modified: 09/26/2002. Accepted: 10/22/2002

Rosala Servn Villegas. Biologist, Instituto Politcnico (IPN), Ciudad de Mxico. M.Sc. in Animal Biology, Universidad Nacional Autnoma de Mxico (UNAM). Investigador Titular, Centro de Investigaciones Biolgicas del Noroeste, S.C. (CIBNOR). Address: Mar Bermejo #195, Playas El Con-

chalito Apdo. Postal 128, La Paz, B.C.S., Mxico, 23090 email: rservin@cibnor.mx Ricardo Aguilar Aguilar. Biologist, Universidad Autnoma de Nuevo Len, Mxico. M.Sc. in Animal Biology, UNAM. Tcnico Acadmico Titular, CIBNOR. e-mail: aguilar6@prodigy.net.mx

Jos L. Martnez Carrillo. Agronomical Engineer, Universidad Autnoma de Sinaloa, Mxico. M.S. and Ph.D. in Entomology, University of California at Riverside. e-mail: jlmc@cirno.inifap.conacyt.mx

Enrique Troyo Dieguez. M.Sc and D.Sc, UNAM. Investigador Titular, CIBNOR. e-mail: etroyo@cibnor.mx Alfredo Ortega. M.Sc. and D. Sc., IPN. Investigador Titular, CIBNOR. e-mail: aortega@cibnor.mx.

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RESUMO Se determinaram os nveis de toxicidade em povoaes de picudo do chile Anthonomus eugenii a tres inseticidas, carbaril, endosulfan e metomil. As povoaes de insetos foram obtidas de trs localidades em Baixa California Sul, Mxico: Benito Jurez (2753N 11346W), San Jos Viejo (2307N, 10943W) e San Juan de los Planes (2358N, 10956W). As CL50 mais elevadas correspondem ao carbaril (178.4,635.4 e 5462.1 gr/ml); endosulfan teve valores intermdios (36.5, 34.1 e 47.1 g/ml); enquanto que os valores mais baixos corresponderam a metomil (1.3, 1.7 e 2.7 g/ml), para Benito Jurez, San Jos Viejo e San Juan de los Planes respectivamente. Se pode concluir que as povoaes de picudo do chile de San Juan de los Planes so resistentes aos trs inseticidas utilizados, tendo uma marcada resistncia a carbaril, em comparao com as povoaes provenientes de San Jos el Viejo e Benito Jurez. Os nveis de susceptibilidade correspondem com as quantidades de inseticidas usados em contra do picudo del chile nas trs localidades de estudo e so uma resposta presso seletiva exercida por estes compostos.

tions, the culture of chili pepper is very successful and is a significant source of employment. In the period 19931997, an annual average of 1600ha were planted, with a production of 28700 metric tons, and an approximate value of MX$ 70 million. However, losses due to pepper weevil damage during 19961997 were estimated at MX$ 10 million (SAGAR, 1997). The effectiveness of several insecticides, either alone or mixed, for the management of the pepper weevil has been assessed in field conditions (Hernndez and Gonzlez, 1992; Pacheco-Covarrubias, 1993; Enrquez and Valenzuela, 1996), and the resistance of the insect to different insecticides has also been estimated using bioassays (Quiones and Flores, 1991; Lpez, 1996). To contribute to the efficient management of pepper weevil, this study assessed the resistance of the insect to three commonly applied insecticides, in three locations of Baja California Sur characterized by their geographic isolation from each other and by the quality of their chili pepper crop, generally exported or traded in other regions of Mexico. There are increasing restrictions for trading products contaminated with insecticides in international markets and a risk posed to domestic consumers by the increasing use of these chemicals. These reasons, combined with the evolution of the pepper weevils resistance to the chemicals and their costs, make it imperative for alternative insect

management programs to be implemented. These programs should include regular assessment of the resistance of pepper weevil populations to different insecticides. The objective of this work was to evaluate the resistance levels of the pepper weevil to the three pesticides most commonly used in the selected locations, by determining LC50 and LC 95 values, confidence limits, and dose-mortality slope values. Material and Methods This study was conducted from December 1997 to March 1998, in the main chili pepper cultivating areas in Baja California Sur. The locations were Benito Jurez (2753N, 11346W) in the municipality of Muleg; San Jos Viejo (2307N, 10943W) in the municipality of Los Cabos; and San Juan de los Planes (2358N, 10956W), in the municipality of La Paz. These areas are geographically isolated from each other by natural mountain barriers and by xerophytic vegetation on non-agricultural land, conforming agricultural valleys or oases. Within each location, parcels of land on which chili peppers (Ancho San Luis variety) are cultivated were selected. In each parcel, 10-12kg of dropped fruit were collected and deposited in plastic containers of 60cm diameter and 25cm depth. The containers were covered with a screen and kept in the laboratory until the adult insects hatched.

These adult insects were collected with a microaspirator and transferred to screened wooden cages of 25 x 25 x 40cm. The most active insects were selected, eliminating those having abnormal mobility. The insects were fed until 4 days old on a diet of fresh cut chili pepper, before being used for the toxicological bioassays with the glass vial technique (Plapp et al., 1987; Cahill and Hackett, 1992). The insecticides assessed were technical grade carbaryl, endosulfan and methomyl, which according to direct surveys of the farmers (published information was not available), are most commonly used against the pepper weevil pest. Five different concentrations with five replicates each were prepared for every insecticide, using acetone as a solvent. Each replicate included a control with acetone only. Five adult insects were introduced in 20ml scintillation vials whose caps were modified to allow the organisms to breathe, and the tubes were inverted to ensure continuous contact between the insects and the insecticides. After 24hr mortality was evaluated, considering as affected or dead any individual that was immobile or that had abnormal mobility. Bioassays were conducted at room temperature, between 22.5 and 28.5C, and relative humidity between 55% and 65%. The dose-mortality response lines for each chemical were obtained using the Probit program (Raymond, 1985).

Results and Discussion The insecticide concentration lethal to 50% of the insects (LC 50), the confidence range at 95% (CR 95%), the lethal concentration at 95% (LC95), the slope of each regression line, and the standard error (SE) for each population from the three studied locations are shown in Table I. For all three pepper weevil populations, methomyl had the highest toxicity, while endosulfan was intermediate and carbaryl was the least toxic. Based on confidence range overlap (Figure 1) it was determined that sensitivity to carbaryl was significantly different among the three populations evaluated. The Benito Jurez weevils were more sensitive than those from San Jos Viejo, which in turn were more sensitive than those from San Juan de los Planes, where resistance to carbaryl was caused by the excessive use of this pesticide for more than 10 years (personal communication from producers), as this was the pesticide most recommended by governmental agencies. The response to methomyl between pepper weevil populations from Benito Jurez and San Juan de los Planes showed a significant difference, but not that between populations from Benito Jurez and San Jos Viejo. With respect to endosulphan, there was no significant difference between the three populations evaluated, according to the overlap in the confidence ranges at the LC 50 level.

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Figure 1. LC50 values obtained with carbaryl, endosulfan and methomyl in three locations of Baja California Sur, Mexico. : Benito Jurez, : San Jos Viejo, : San Juan de los Planes. * Maximum level for carbaryl concentration in San Juan de Los Planes was 15.144 g/ml.

Based on the results, it is evident that of the three pepper weevil populations evaluated in Baja California Sur, that from the agricultural zone of San Juan de los Planes has the greatest pesticide resistance problem, due to the selective pressure exerted by the compounds used to combat them. The populations of San Jos Viejo showed an intermediate response to the insecticides, while the lowest resistance was observed in the Benito Jurez populations. Carbaryl was the pesticide mostly used in the studied areas, and it was found that the doses required in San Juan de los Planes are 30 times higher than in Benito Jurez, and 8.5 times higher than in San Jos. Therefore, it is recommended that carbaryl no longer be used to control the pepper weevil in any of the men-

tioned areas. The susceptibility of the three populations to endosulfan and methomyl is quite similar. Therefore, it is suggested that if these pesticides are used, precaution should be taken to avoid repetition of the carbaryl problem. Other strategies should be implemented in order to

reduce selection pressure on the pepper weevil and other insect pests in this regin. These may include planting dates, destruction of plant residues, establishment of host free periods, alternative chemical or biological insecticides, and biological control. There is little information published on pepper weevil resistance to the insecticides used in this study. Genung and Osaki (1972) refer methomyl, carbaryl and endosulfan resistance when conducting field assessments with these products. In test fields and commercial parcels Rolston (1977) found pepper weevils resistant to endosulfan and methomyl, reporting mortalities of 15% and 65%, respectively. In the agricultural valley of La Paz, Baja California Sur, Servn and Aguilar (2000) found insects resistant to carbaryl with an LC 50 of 3565.7g/ml, and to endosulfan, with an LC50 of 131.8g/ ml, consistent with the results presented in this study. According to the insecticide application record in the studied zone, the situation in San Juan de los Planes is critical. It is important to consider that these pesticides are highly toxic to organisms other than the pepper weevil. The use of methomyl and carbaryl is restricted, as they are highly poisonous to human beings and other animals by inhalation or contact. In addition, some pesticides are unstable; 50% of the applied endosulfan residue is lost or de-

graded within three to seven days. The control alternatives for the pepper weevil are not encouraging; therefore, it is important to regularly assess the reincidence of this pest and to manage pesticide use in order to avoid increased resistance.
ACKNOWLEDGEMENTS

The authors thank the collaboration of Dinora Romero, Instituto Nacional de Investigaciones Forestales y Agropecuarias (INIFAP) and Amado Cota, CIBNOR, Sociedad Civil. This study was financed by the Consejo Nacional de Ciencia y Tecnologa, Secretara de Educacin Pblica, Centro de Investigaciones Biolgicas del Noroeste , through Project AGEC-2, and by the Sistema de Investigacin del Mar de Corts through Project Code 990106010.
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TABLE I TOXICITY OF INSECTICIDES AGAINST Anthonomus eugenii IN THREE LOCATIONS OF BAJA CALIFORNIA SUR, MEXICO Location Benito Jurez San Jos Viejo San Juan de los Planes Pesticide Carbaryl Endosulfan Methomyl Carbaryl Endosulfan Methomyl Carbaryl Endosulfan Methomyl LC50 (g/ml) 178.4 36.5 1.3 635.4 34.1 1.7 5462.1 47.1 2.7 95% CR 104.8 23.3 0.6 327.9 2.2 0.9 3135.7 17.9 2.2 284.6 54.4 1.8 1420.1 67.9 2.4 15144 108.3 3.3 LC95 (g/ml) 3585.8 345.5 11.7 31541.6 1277.4 10.9 132089 5059.6 8.1 Slope 1.26 1.68 1.77 0.97 1.04 2.02 1.18 0.81 3.46 SE 0.206 0.298 0.507 0.188 0.387 0.51 0.282 0.24 0.503

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Campo Experimental Norte de Guanajuato / Campo Experimental Bajo. SARH. INIFAP. PIAFEG. Mxico. 6pp. Cahill MR, Hackett B (1992) Insecticidal activity and expression of pyrethroid resistance in Bemisia tabaci using a glass vial bioassay. Brighton Crop Protection Conference Pest and Diseases. 3C-15: 251-256. Enrquez J, Valenzuela P (1996) Evaluacin de insecticidas para el control del picudo del chile Anthonomus eugenii Cano. Memorias VI Congreso Latinoamericano y XXXI Congreso Nacional de Entomologa. Mrida, Yucatn, Mxico. pp. 119-120. Genung WG, Ozaki HY (1972) The pepper weevil Anthonomus eugenii Cano on the lower Florida east coast . Belle Glade. AREC Mimeo Report EV-1972:2. Hernndez J, Gonzlez R (1992) Efecto de siete insecticidas sobre el control de picudo del

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Cah. ORSTOM, Sr. Ent. Med. Parasitol. 22: 117-121. Riley DG, King EG (1994) Biology and management of pepper weevil Anthonomus eugenii Cano (Coleoptera: Curculionidae): A review. Entomol. (Trends in Agril.Sci) 2 : 109121. Rolston LH (1977) Insecticide tests in laboratory and field against the pepper weevil, Anthonomus eugenii Cano. J. Georgia Entomol. Soc. 12: 117-120. SAGAR (1997) Programa fitosanitario de contingencia para el control del picudo o barrenillo del chile (Anthonomus eugenii). Comit Regional de Sanidad Vegetal La Paz-Los Cabos. SAGAR. Mxico. 13 pp. Servn R, Aguilar R (2000) Bioensayos toxicolgicos en picudo del chile Anthonomus eugenii Cano (Coleoptera: Curculionidae) por exposicin residual, en Baja California Sur, Mxico. Fol. Entomol. Mex. 109: 117-118.

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