Laboratory Rearing and Vital Statistics of Diabrotica Speciosa and Diabrotica Viridula (Coleoptera: Chrysomelidae), Two Species of South American Pest Rootworms
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ISSN 0373-5680 Rev. Soc. Entornol. Argent. 60 (1-4): 239-248, 2001 Laboratory rearing and vital statistics of Diabrotica speciosa and Diabrotica viridula (Coleoptera: Chrysomelidae), two species of South American pest rootworms C.A~BRERAlVALSH, Guillermo J.Cabrera Walsh, Guillermo J. South American Biological Control Laboratory, Agricultural Counselor, Agricultural Research Service Laboratory,U.S. Embassy. Unit 4325, APO AA 34034-0001. USA Bolivar 1559. 1686 Hurlingham, Argentina; e-mail: [email protected] • ABSTRACT. Methods for rearing Diabrotica speciosa (Germar) and D. viridula (F.) in the laboratory, and their vital statistics are described. These Luperini are pests of several crops in South and Central America, and there was no pre vious work describing suitable methods for obtaining the numbers required for the mass production of parusitoids, assessing pathogens, or conducting host specificity studies. Handling of immatures and adults was reduced to a minimum, and materials used were inexpensive, reducing costs, labor time, and mortality related to manipulation. Larvae were reared on maize seedlings in a soil mixture, and adults were fed a dry diet. Oviposition was obtained in cups with corn seedlings wrapped in black cloth; eggs were harvested by rinsing them off the cloths with water, and sown by squirting them into maize cultures with plastic pipettes. Percentage of hatch, survival from egg to adult, and number of eggs laid per female were the highest cited yet forD. speciosa %,25.9%, %, (D. speciosa: 92.6 and 1164 respectively;D. viridula: 91 %,25 and "1271 respectively), and sirnilar for both species. D. viridula proved to be a more vigorous species than D. speciosa, with a longer average Iife span, and a slightly higher egg production, at least in present laboratory conditions. KEY WORDS. Diabrotica speciosa ..Diabrotica viridula. Laboratory rearing. Rootworms. • RESUMEN. Cria enlaboratorio y estadisticas vitales de Diabrotica speciosa y Diabrotica viridula (Coleoptera: Chrysomelidae), dos especies de vaquitas sudamericanas plagas de cultivos, Se describen metodos para la cria en labora torio deDiabrotica speciosa (Germar) y Diabrotica viridula (F.)y sus estadisticas vitales. Estos Luperini son plagas de varios cultivos en America del Sur y Central, y no habia trabajos previos describiendo metodos adecuados para obtener las cantidades de adultos y larvas necesarias para crías rnasivas de enemigos naturales, estudios de patógenos,o pruebas de especificidad de enemigos naturales. EI manipuleo de inmaduros y adultos, y su mortalidad asociada, fueron reducidos alminimo. Los materiales util izados fueron de bajo costo, y se redujeron las horas hombre. Las larvas se alimentaron de plantulas de maiz y los adultos de una dieta seca. Se obtuvieron puestas en recipientes con plantulas de maíz envueltas con rectangulos de tela negra hu medecida: los huevos se cosecharon mediante chorros de agua de los trozos de tela y se sembraron en los cultivos de rnaiz mediante pipetas plasticas .. EI porcentaje de eclosión, supervivencia de huevo a adulto, y promedio de hue % vos por hembra fueron simi lares en ambas especies (D. speciosa: 92,6 , %, 25,9 Y 1164 respectivamente: D. viridula: 91%, 25% Y 1271 respectiva mente), y los mas altos obtenidos en laboratorio hasta ahora para D. speciosa. 239 Rev. Soc. Eniomol. Argent. 60 (1-4)/ 2001 En las condiciones de laboratorio utilizadas,D. viridula resultó una especie mas vigorosa queD. speciosa, con una rnayor longevidad y producción de huevos. PALABRAS CLAVE.Diabrotica speciosa. Diabrotica viridula. Cria en laboratorio. Chrysomel idae plagas. INTROD'UCTI()N squash leaves and fruit, corn seedlings), or both (Mendoza & Peters, 1963; Hovve & George, The subtribe Diabroticina has polyphagous 1966; Chalfant & Mitchell, 1968; Cuthbert et al., and 01 igophagous species that have been recor 1968; Skelton Bx Hunter, 1970; Branson et al., ded to feed on many food, ornarnental, and wild "1975; Jackson, 1986; Schalk, 1986; Campbell & plants. Of these, the genus Diabrotica Chevrolat Jackson, 1987; Branson 8x Jackson, 1988; Branson has the greater nurnber of pest species, including et al., 1988). Several attempts to rear immature some of the most important rO\1\I and orchard stages of Oiabrotica on artificial diets have been crops pests of the Americas. In South America, successful (Sutter et al., 1971; Rose & McCabe, the most common and pestiferous species is 1973; Marrone et al., 1985; Avila et al., 2000). Oiabrotica speciosa (Germar). The adults are However, they have proven less effective and blamed for important damage in maize, as part more labor intensive than the live hosts methods, of the kernels do not mature because they eat so Diabroticites are, but exceptionally, at present the silks; cucurbits, where they eat the flowers reared on live hosts. and young fruit; heavy defoliation in soya and The objectives of this work are to report the re damage to the tender parts of al most every crop sults of the methods developed at the ARS-USDA (Christensen, 1943; Silva et al., 1968; link & Cos South American Biological Control laboratory ta, 1978). They also may transmit bacterial wilt in (SABCl), Hurlingham, Argentina, for rearing D. cucurbits (Cabrera VValsh, unpublished data). speciosa and O. viridula, and their vital statistics, Although larval damage to crops has not been with notes on thei I' development and biology. evaluated rigorously, there is sound evidence that they seriously damage corn, wheat, potatoes and peanuts (Sarasola et al., 1980; Gassen, 1984). MATERIAL AND METHODS Oiabrotica speciosa belongs to the fucata group, which contains generally multivoltine, polypha Field Collections. The founding beetles of the gous species, and none of thern are known to have LJ. speciosa and 0'. viridula colonies were adults diapausing eggs, but rather overwinter as adults collected in the field throughout northeastern and (Krysan & Srnith, 1987). The other group within the central Argentina. Eggs were harvested from the genus, the virgifera group, includes the worst pests field collected adults, reared in the laboratory, of maize in North America, based on annual losses and the adults mixed with the laboratory colony, in yields and control costs (Metcalf, 1986). This to maintain a diverse gene pool. group generally shows a stricter stenophagy, es The beetles were usually collected off fol iage pecially of larval hosts, diapausing eggs, and by sweeping. Corn silks or flower heads were univoltinism (Krysan & Smith, 1987). The sole knocked over the edge of a plastic funnel atta South American species of this group that may ar ched to a plastic container. In delicate crops, such guably be considered a pest, is Oiabrotica viridula as lettuce, where sweeping would cause extensive (F.), reported to cause significant darnage on maize damage, they vvere collected with hand aspirators. (Harries, 1975; Olalquiaga, 1980), and to trans Battery-operated aspirators a'nd a D-vac also ha mit maize chlorotic rnottle virus in Peru (Reyes & ve been tried, but they have not proven useful, Castillo,1988). because they clog with leaves and petals easily. Stud ies on these insects have led to the deve Together with these methods, polyester cloths lopment of several laboratory rearing procedures. sprayed vvith alcoholic Cayaponia spp. root or Most of these methods involve reari ng larvae on a berry extracts were deployed at every collection si live host, typically corn seedlings, and feeding the te, to attract beetles to them while the workers co adults meridic diets, fresh food (bean leaves, llected on the plants (Cabrera Walsh, unpublished 240 CABRERA WALSH, G. J. Laboratory rearing and vital statistics of Diabrotica data). When collecting was complete, the cloths Table 1. Diets used for Diabroiica speciosa, andD. tnri were shaken into a sweeping net, which was dula (for 1Kg of diet) emptied of its contents of beetles into a cage, or Ingredients Amounts separated by species on the spot with aspirators. Soy flour 54.5 g Wheat germ 54.5 g laboratory Handling. T\I\lO sizes of cylindrical Casein 70 g cages with sleeves (1.5 and -19 liters) were used AlphaceJ 82 g for rearing adults and collecting their parasitoids. Sucrose 145 g The small containers can hold ca. 150 beetles, Brewers yeast 22 g while the large ones up to 1500 beetles. Both Glicerine 60 g had a size 12 plastic rnesh attached to the botton]' Wesson salts mix 5.5 g through which ernerging parasitoids fell onto a Vanderzant vitamins mix 5.5 g moist sand bed. Adults were fed a diet based on Cholesterol 1.0 g Campbell & Jackson (1987), to which antimicro Methylparaben 0.65 ga bials were added (Table ~I). Water was provided Sorbic acid 0.92 ga from 45 rnl plastic cups with cotton wool wicks Acetic acid 2.7ml through the Iids. Distilled water 500 ml Rearing. Oviposition was obtained in 42-ml a dissolved together in 8rnl of ethanol. cups with moist squares of folded black cotton/polyester cloth, with maize seed lings roo- Blend the mixture with part of the water after each in gredient is added. Squeeze 5-mrn thick, 2-cm wide ted amidst the folds. Females laid their eggs in the rows on successive layers of aluminum foil and freeze. folds. The seedlings were obtained by incubating Thaw in microwave and spread on shallow lid. Renew 40-.50 kernels at 25°C in O.S-liter plastic contai every week-Ill d or when consumed. ners with 3 crn of moist oven-steril ized sand. When the seedlings reached 3-7 CITl in height, the 100 eggs, based on the average volume of five sand was washed away, and the seed ling roots (5 previously counted samples of 1- to 3-d-old eggs. 7 per cup) wrapped in the cloths, pressed into The samples were allowed to settle for some 30 s, the cups, and moistened thoroughly. Although and tapped to eliminate air bubbles. The same Diabroticites oviposit on several moist substra procedure was used for O.