Toxicity and sublethal effects of six insecticides to last instar larvae and adults of the biocontrol agents Chrysoperla carnea (Stephens) (Neuroptera: Chrysopidae) and Adalia bipunctata (L.) (Coleoptera: Coccinellidae)

To further develop Integrated Pest Management (IPM) strategies against crop pests, it is important to evaluate the effects of insecticides on biological control agents. Therefore, we tested the toxicity and sublethal effects (fecundity and fertility) of flonicamid, flubendiamide, metaflumizone, spirotetramat, sulfoxaflor and deltamethrin on the natural enemies Chrysoperla carnea and Adalia bipunctata. The side effects of the active ingredients of the insecticides were evaluated with residual contact tests for the larvae and adults of these predators in the laboratory. Flonicamid, flubendiamide, metaflumizone and spirotetramat were innocuous to last instar larvae and adults of C. carnea and A. bipunctata. Sulfoxaflor was slightly toxic to adults of C. carnea and was highly toxic to the L4 larvae of A. bipunctata. For A. bipunctata, sulfoxaflor and deltamethrin were the most damaging compounds with a cumulative larval mortality of 100%. Deltamethrin was also the most toxic compound to larvae and adults of C. carnea. In accordance with the results obtained, the compounds flonicamid, flubendiamide, metaflumizone and spirotetramat might be incorporated into IPM programs in combination with these natural enemies for the control of particular greenhouse pests. Nevertheless, the use of sulfoxaflor and deltamethrin in IPM strategies should be taken into consideration when releasing either of these biological control agents, due to the toxic behavior observed under laboratory conditions. The need for developing sustainable approaches to combine the use of these insecticides and natural enemies within an IPM framework is discussed.

Ecotoxicity of insecticides of frequent use in tomatoes on Trichogramma achaeae (Hymenoptera: Trichogrammatidae)

Since Tuta absoluta(Meyrick) (Lepidoptera: Gelechiidae) was detected in 2006 as a new pest in tomato crops in Spain, several natural enemies have been reported tocontrol this pest. In biological control programs, the native parasitoid Trichogramma achaeae Nagaraja&Nagarkatti (Hymenoptera: Trichogrammatidae) is used against T.absoluta. However, the most common control practice is based on use of pesticides,and in the frame of Integrated Pest Management (IPM) programs, the knowledge on the activity of insecticides towards beneficial insects is needed for its joint use. In thiswork, we evaluated lethal and sublethal effects of insecticides commonly applied on tomato crops on adults of T. achaeae. Pesticides were sprayed on tomato plants or T. Absoluta eggs till run off at their maximum field recommended concentration. Mortality was scored after 24, 48 and 72 hours, as well as beneficial capacity and percentage of emergence.

Fenología y capacidad de dispersión de Monochamus galloprovincialis (Olivier 1795) en la Península Ibérica = Phenology and dispersal ability of Monochamus galloprovincialis (Olivier 1795) at Iberian Peninsula

La aparición y avance de la enfermedad del marchitamiento del pino (Pine Wilt Desease, PWD), causada por Bursaphelenchus xylophilus (Nematoda; Aphelenchoididae), el nematodo de la madera del pino (NMP), en el suroeste de Europa, ha puesto de manifiesto la necesidad de estudiar la fenología y la dispersión de su único vector conocido en Europa, Monochamus galloprovincialis (Col., Cerambycidae). El análisis de 12 series de emergencias entre 2010 y 2014, registradas en Palencia, València y Teruel, con material procedente de diversos puntos de la península ibérica, demostró una alta variabilidad en la fenología de M. galloprovincialis y la divergencia térmica respecto de las poblaciones portuguesas. Para éstas, el establecimiento de los umbrales térmicos de desarrollo de las larvas post-dormantes del vector (12,2 y 33,5ºC) permitió la predicción de la emergencia mediana para la fecha en la que se acumulaban de 822 grados-día. Ninguna de las series analizadas en este trabajo necesitó de dichos grados-día estimados para la emergencia mediana. Asimismo, la emergencia se adelantó en las regiones más calurosas, mientras que se retrasó en las zonas más templadas. Más allá de la posible variabilidad entre poblaciones locales peninsulares, se detectaron indicios de que la diferencia en la acumulación de calor durante el otoño puede afectar el grado de maduración de las larvas invernantes, y su posterior patrón temporal de emergencia. Por último, también fueron observados comportamientos de protandria en las emergencias. Respecto a la fenología de su vuelo, entre los años 2010 y 2015, fueron ejecutados un total de 8 experimentos de captura de M. galloprovincialis mediante trampas cebadas con atrayentes en diferentes regiones (Castellón, Teruel, Segovia y Alicante) permitiendo el seguimiento del periodo de vuelo. Su análisis permitió constatar la disminución de las capturas y el acortamiento del periodo de vuelo con la altitud, el inicio del vuelo en el mes de mayo/junio a partir de los 14ºC de temperatura media diaria, la influencia de las altas temperaturas en la disminución de las capturas estivales (potencial causante de perfiles bimodales en las curvas de vuelo en las zonas menos frías), la evolución de la proporción de sexos a lo largo del periodo de vuelo (que muestra una mayor captura de hembras al inicio y de machos al final) y el comportamiento diurno y ligado a las altas temperaturas del vuelo circadiano del insecto. Dos redes de muestreo sistemático de insectos saproxílicos instaladas en la Comunitat Valencia (Red MUFFET, 15 parcelas, año 2013) y en Murcia (Red ESFP, 20 parcelas, años 2008-2010) permitieron el estudio de la comunidad de insectos relacionada con M. galloprovincialis. Cada una de las parcelas contaba con una trampa cebada con atrayentes y una estación meteorológica. El registro de más de 250 especies de coleópteros saproxílicos demostró el potencial que tiene el empleo de redes de trampas vigía para la detección temprana de organismos exóticos, además de permitir la caracterización y evaluación de las comunidades de entomofauna útil, representando una de las mejores herramientas de la gestión integrada de plagas. En este caso, la comunidad de saproxílicos estudiada mostró ser muy homogénea respecto a la variación ambiental de las zonas de muestreo, y que pese a las pequeñas variaciones entre las comunidades de los diferentes ecosistemas, el rol que M. galloprovincialis desempeña en ellas a lo largo de todo el gradiente estudiado es el mismo. Con todo, el análisis mediante redes de interacción mostró su relevancia ecológica al actuar de conector entre los diferentes niveles tróficos. Por último, un total de 12 experimentos de marcaje-liberación-recaptura desarrollados entre 2009 y 2012 en Castellón, Teruel, Valencia y Murcia permitieron evaluar el comportamiento dispersivo de M. galloprovincialis. Las detecciones mediante trampas cebadas de los insectos liberados se dieron por lo menos 8 días después de la emergencia. La abundancia de población pareció relacionada con la continuidad, la naturalización de la masa, y con la afección previa de incendios. La dispersión no estuvo influida por la dirección ni la intensidad de los vientos dominantes. La abundancia de material hospedante (en lo referente a las variables de masa y a los índices de competencia) influyó en la captura del insecto en paisajes fragmentados, aunque la ubicación de las trampas optimizó el número de capturas cuando se ubicaron en el límite de la masa y en zonas visibles. Por último también se constató que M. galloprovincialis posee suficiente capacidad de dispersión como para recorrer hasta 1500 m/día, llegando a alcanzar distancias máximas de 13600m o de 22100 m. ABSTRACT The detection and expansion of the Pine Wilt Desease (PWD), caused by Bursaphelenchus xylophilus (Nematoda; Aphelenchoididae), Pine Wood Nematode (PWN), in southwestern Europe since 1999, has triggered off the study of the phenology and the dispersion of its unique vector in the continent, Monochamus galloprovincialis (Coleoptera, Cerambycidae). The analysis of 12 emergence series between 2010 and 2014 registered in Palencia, Teruel and Valencia (Spain), registered from field colonized material collected at several locations of the Iberian Peninsula, showed a high variability in the emergence phenology of M. galloprovincialis. In addition, these patterns showed a very acute thermal divergence regarding a development model fitted earlier in Portugal. Such model forecasted the emergence of 50% of M. galloprovincialis individuals in the Setúbal Peninsula (Portugal) when an average of 822 degree-days (DD) were reached, based on the accumulation of heat from the 1st of March until emergence and lower and upper thresholds of 12.2 ºC and 33,5 °C respectively. In our results, all analyzed series needed less than 822 DD to complete the 50% of the emergence. Also, emergency occurred earlier in the hottest regions, while it was delayed in more temperate areas. Beyond the possible variability between local populations, the difference in the heat accumulation during the fall season may have affected the degree of maturation of overwintering larvae, and subsequently, the temporal pattern of M. galloprovincialis emergences. Therefore these results suggest the need to differentiate local management strategies for the PWN vector, depending on the location, and the climatic variables of each region. Finally, protandrous emergence patterns were observed for M. galloprovincialis in most of the studied data-sets. Regarding the flight phenology of M. galloprovincialis, a total of 8 trapping experiments were carried out in different regions of the Iberian Peninsula (Castellón, Teruel, Segovia and Alicante) between 2010 and 2015. The use of commercial lures and traps allowed monitoring of the flight period of M. galloprovincialis. The analyses of such curves, helped confirming different aspects. First, a decline in the number of catches and a shortening of the flight period was observed as the altitude increased. Flight period was recorded to start in May / June when the daily average temperature went over 14 ° C. A significant influence of high temperatures on the decrease of catches in the summer was found in many occasions, which frequently lead to a bimodal profile of the flight curves in warm areas. The evolution of sex ratio along the flight period shows a greater capture of females at the beginning of the period, and of males at the end. In addition, the circadian response of M. galloprovincialis to lured traps was described for the first time, concluding that the insect is diurnal and that such response is linked to high temperatures. Two networks of systematic sampling of saproxylic insects were installed in the Region of Valencia (Red MUFFET, 15 plots, 2013) and Murcia (Red ICPF, 20 plots, 2008-2010). These networks, intended to serve the double purpose of early-detection and long term monitoring of the saproxylic beetle assemblies, allowed the study of insect communities related to M. galloprovincialis. Each of the plots had a trap baited with attractants and a weather station. The registration of almost 300 species of saproxylic beetles demonstrated the potential use of such trapping networks for the early detection of exotic organisms, while at the same time allows the characterization and evaluation of useful entomological fauna communities, representing one of the best tools for the integrated pest management. In this particular case, the studied community of saproxylic beetles was very homogeneous with respect to environmental variation of the sampling areas, and despite small variations between communities of different ecosystems, the role that M. galloprovincialis apparently plays in them across the studied gradient seems to be the same. However, the analysis through food-webs showed the ecological significance of M. galloprovincialis as a connector between different trophic levels. Finally, 12 mark-release-recapture experiments were carried out between 2009 and 2012 in Castellón, Teruel, Valencia and Murcia (Spain) with the aim to describe the dispersive behavior of M. galloprovincialis as well as the stand and landscape characteristics that could influence its abundance and dispersal. No insects younger than 8 days were caught in lured traps. Population abundance estimates from mark-release-recapture data, seemed related to forest continuity, naturalization, and to prior presence of forest fires. On the other hand, M. galloprovincialis dispersal was not found to be significantly influenced by the direction and intensity of prevailing winds. The abundance of host material, very related to stand characteristics and spacing indexes, influenced the insect abundance in fragmented landscapes. In addition, the location of the traps optimized the number of catches when they were placed in the edge of the forest stands and in visible positions. Finally it was also found that M. galloprovincialis is able to fly up to 1500 m / day, reaching maximum distances of up to 13600 m or 22100 m.

Compatibilidad de Eretmocerus mundus Mercet (Hymenoptera: Aphelinidae) y Amblyseius swirskii Athias-Henriot (Acari: Phytoseiidae), importantes enemigos naturales de la mosca blanca Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) en cultivos hortícolas, con nuevas barreras físicas selectivas y modernos plaguicidas

Los programas de Gestión Integrada de Plagas (GIP) promueven el uso de estrategias de control que sean respetuosas con el medio ambiente, sin embargo el uso de insecticidas en los cultivos hortícolas sigue siendo necesario para el control de determinadas plagas, como es el caso de la mosca blanca Bemisia tabaci (Gennadius). Por ello, el objetivo de esta tesis es el estudio de la integración de las tres estrategias de control más empleadas hoy en día para el control de plagas: el control biológico, el físico y el químico. Una primera parte de este trabajo ha consistido en el estudio de los efectos letales y subletales de once insecticidas, aplicados a la dosis máxima de campo, sobre los enemigos naturales Eretmocerus mundus Mercet y Amblyseius swirskii Athias-Henriot, mediante ensayos de laboratorio y persistencia (laboratorio extendido). Para la evaluación de la toxicidad de los insecticidas sobre los estados de vida más protegidos de estos enemigos naturales, se trataron bajo la Torre de Potter las pupas de E. mundus y los huevos de A. swirskii. Además, se llevaron a cabo ensayos de contacto residual para determinar los efectos letales y subletales de estos insecticidas sobre el estado adulto de ambas especies de enemigos naturales. Para ello, los pesticidas se aplicaron sobre placas de cristal (laboratorio) o sobre plantas (laboratorio extendido: persistencia). Los resultados mostraron que los insecticidas flonicamida, flubendiamida, metaflumizona, metoxifenocida, spiromesifen y spirotetramat eran compatibles con el estado de pupa de E. mundus (OILB 1: Inocuos). Sin embargo, abamectina, deltametrina y emamectina fueron categorizadas como ligeramente tóxicas (OILB 2) al causar efectos deletéreos. Los dos pesticidas más tóxicos fueron spinosad y sulfoxaflor, los cuales redujeron significativamente la emergencia de las pupas tratadas (OILB 4: Tóxicos). Flonicamida, flubendiamida, metoxifenocida y spiromesifen fueron compatibles con el estado adulto de E. mundus (OILB 1: Inocuos). Abamectina, deltametrina, emamectina, metaflumizona y spiromesifen pueden ser recomendados para su uso en programas de GIP, si se usan los plazos de seguridad apropiados, de acuerdo con la persistencia de cada uno de estos insecticidas, antes de la liberación del enemigo natural. Al contrario, spinosad y sulfoxaflor no resultaron ser compatibles (OILB D: Persistentes), aunque la realización de ensayos adicionales es necesaria para ver los efectos de los mismos en campo. Todos los insecticidas estudiados, excepto el spirotetramat (OILB 2: Ligeramente tóxico), fueron selectivos para el estado de huevo de A. swirskii (OILB 1: Inocuos). Flonicamida, flubendiamida, metaflumizona, metoxifenocida, spiromesifen, spirotetramat y sulfoxaflor, fueron compatibles con el estado adulto de A. swirskii (OILB 1: Inocuos). Abamectina, deltametrina, emamectina y spinosad pueden ser recomendados para su uso en programas de GIP, si se usan los plazos de seguridad apropiados, de acuerdo con la persistencia de cada uno de estos insecticidas, antes de la liberación del enemigo natural. Entre las nuevas estrategias de la GIP, los plásticos y mallas fotoselectivas han demostrado ser una herramienta importante para el control de plagas y enfermedades en cultivos hortícolas protegidos. Por ello, en una segunda parte de este trabajo, se estudiaron tanto los efectos directos, como la combinación de efectos directos y mediados por planta y plaga de ambientes pobres en luz UV, en presencia o ausencia del Virus del rizado amarillo del tomate (TYLCV), sobre E. mundus. En primer lugar, se realizó un ensayo al aire libre para la evaluación de la capacidad de vuelo de E. mundus en cajas tipo túnel (1 x 0,6 x 0,6 m) cubiertas con distintas barreras absorbentes de luz UV. Se detectó un efecto directo en la capacidad de orientación de E. mundus, debido a que este parasitoide utiliza estímulos visuales para localizar a sus huéspedes, únicamente en las barreras que bloqueaban más del 65% de la luz UV (malla G). En segundo lugar, bajo condiciones de invernadero, se evaluó la combinación de efectos directos y mediados por planta y plaga sobre E. mundus, usando plantas de tomate sanas o infectadas con el TYLCV y cajas (30 x 30 x 60 cm) cubiertas con los distintos plásticos fotoselectivos. En este caso, no se observó ningún efecto en la capacidad benéfica del parasitoide cuando este estaba en contacto con plantas de tomate infestadas con ninfas de B. tabaci, lo que demuestra que este insecto usa estímulos táctiles para encontrar a sus huéspedes a cortas distancias. Además, las diferentes condiciones de radiación UV estudiadas tuvieron cierto impacto en la morfología, fisiología y bioquímica de las plantas de tomate, infestadas o no con el virus de la cuchara, detectándose pequeñas alteraciones en alguno de los parámetros estudiados, como el peso fresco y seco, el contenido en H y el espesor de las cutículas y de las paredes celulares de la epidermis foliar. Por último, no se observaron efectos de la radiación UV mediados por planta, ni en B. tabaci ni en su parasitoide, E. mundus. En una tercera parte, se evaluaron los efectos de una malla tratada con bifentrin sobre ambos enemigos naturales, en ensayos de laboratorio, semicampo y campo. Las mallas tratadas fueron diseñadas originariamente para el control de mosquitos vectores de la malaria, y actualmente se está trabajando para su uso en agricultura, como una nueva estrategia de control de plagas. En ensayos de laboratorio, cuando adultos de E. mundus y A. swirskii se expusieron por contacto durante 72 horas con la malla tratada (cajas de 6 cm diámetro), se registró una alta mortalidad. Sin embargo, en el ensayo de preferencia, estos enemigos naturales no fueron capaces de detectar la presencia de bifentrin y, en aquellos individuos forzados a atravesar la malla tratada, no se observó mortalidad a corto plazo (72 horas). En estudios de semicampo, llevados a cabo bajo condiciones de invernadero en cajas de 25 x 25 x 60 cm de altura, la capacidad benéfica de E. mundus no se vio afectada. Finalmente, en ensayos de campo llevados a cabo en invernaderos comerciales (4000m2) en Almería, A. swirskii no se vio afectado por la presencia en el cultivo de la malla tratada con bifentrin y los niveles de infestación de B. tabaci y F. occidentalis detectados bajo dicha malla, fueron inferiores a los del control. Por último, se ha evaluado la composición de la microflora bacteriana de tres especies de parasitoides, E. mundus, Eretmocerus eremicus Rose & Zolnerowich y Encarsia formosa Gahan, y la influencia de la misma en su susceptibilidad a insecticidas. Se llevó a cabo una extracción total de ADN de los insectos y la región variable V4 del ARNr se amplificó usando cebadores universales bacterianos. Para identificar las secuencias de los géneros bacterianos presentes en los parasitoides, se realizó una Next Generation sequencing (Illumina sequencing). Una vez identificados los géneros bacterianos, el gen ADNr 16S de las Actinobacterias se amplificó del ADN extraído de los insectos, usando cebadores universales bacterianos y específicos de Actinobacterias, y los productos de la Nested PCR fueron clonados para identificar todas las especies del género Arthrobacter. Tres bacterias (A. aurescens Phillips, A. nicotinovarans Kodama, Yamamoto, Amano and Amichi y A. uratoxydans Stackebrandt, Fowler, Fiedler and Seiler), próximas a las especies de Arthrobacter presentes en los parasitoides, se obtuvieron de la colección bacteriana del BCCMTM/LMG y se midió su actividad esterasa. Finalmente, se realizaron ensayos con antibióticos (tetraciclina) y de contacto residual con insecticidas (abamectina) para determinar la influencia de las especies de Arthrobacter en la susceptibilidad de E. mundus a insecticidas. Los resultados muestran que este género bacteriano puede afectar a la toxicidad de E. mundus a abamectina, mostrando la importancia de la comunidad microbiana en enemigos naturales, factor que debe ser considerado en los estudios de evaluación de los riesgos de los insecticidas. ABSTRACT Integrated Pest Management (IPM) programs promote the use of control strategies more respectful with the environment; however the use of insecticides in vegetable crops is still needed to control certain pests, such as the whitefly Bemisia tabaci (Gennadius). Therefore, the objective of this work is to study the integration of the three most commonly used pest control strategies nowadays: biological, physical and chemical control. Firstly, the lethal and sublethal effects of eleven insecticides, applied at their maximum field recommended concentration, on the parasitic wasp Eretmocerus mundus Mercet and the predator Amblyseius swirskii Athias-Henriot has been assessed in the laboratory and in persistence tests (extended laboratory). To test the effects of pesticides on the most protected life stage of these natural enemies, E. mundus pupae and A. swirskii eggs were sprayed under a Potter precision spray tower. Laboratory contact tests were therefore conducted to determine the lethal and sublethal effects of these pesticides on the adult stage of these natural enemies. In the residual contact tests the pesticides were applied on glass plates (laboratory) or plants (extended laboratory: persistence). The study showed that the insecticides flonicamid, flubendiamide, metaflumizone, methoxyfenozide, spiromesifen and spirotetramat were selective for E. mundus pupae (IOBC 1: Harmless). Nevertheless, abamectin, deltamethrin and emamectin were categorized as slightly harmful (IOBC 2) due to the deleterious effects caused. The two most harmful pesticides were spinosad and sulfoxaflor, which significantly reduced the adult emergence from treated pupae (IOBC 4: Harmful). Flonicamid, flubendiamide, methoxyfenozide and spiromesifen were compatible with E. mundus adults (IOBC 1: Harmless). Base on the duration of the harmful activity, abamectin, deltamethrin, emamectin, metaflumizone and spirotetramat could be recommended for use in IPM programs if appropriate safety deadlines are used before the natural enemy release. On the contrary, spinosad and sulfoxaflor were not compatible (IOBC D: persistent), although additional studies are required to determine their effects under field conditions. All the pesticides tested, except spirotetramat (IOBC 2: Slightly harmful), were selective for A. swirskii eggs (IOBC 1: Harmless). Flonicamid, flubendiamide, metaflumizone, methoxyfenozide, spiromesifen, spirotetramat and sulfoxaflor were compatible with A. swirskii adults (IOBC 1: Harmless). However, abamectin, deltamethrin, emamectin and spinosad could be recommended for use in IPM programs if appropriate safety deadlines are used before the natural enemy release. Among new IPM strategies, UV-absorbing photoselective plastic films and nets have been shown to be an important tool for the control of pests and diseases in horticultural protected crops. Because of that, we secondly studied the plant and pest insect-mediated and/or the direct effects on E. mundus under different UV radiation conditions, in presence or absence of the Tomato Yellow Leaf Curl Virus (TYLCV). In the first experiment, performed outdoors, the flight activity of E. mundus was studied in one-chamber tunnels (1 x 0.6 x 0.6 m) covered with different photoselective barriers. Because E. mundus uses visual cues for host location at a long distance, a direct effect on its host location ability was detected, but only in the UV-absorbing barriers blocking more than 65% of the UV light (G net). In a second experiment, the direct and plant and pest insect-mediated effects of different UV radiation conditions on E. mundus were studied, inside cages (30 x 30 x 60 cm) covered with the different UVplastic films and under greenhouse conditions, using healthy or TYLCV-virus infected tomato plants. In this case, not any effect on the beneficial capacity of this parasitoid was detected, proving that he uses tactile cues at a short distance of the host. Moreover, the different UV radiation conditions studied had a certain direct impact in the morphology, physiology and biochemistry of tomato plants infested or not with the TYLCV, and small alterations in some parameters such as fresh and dry weight, H percentage and cuticle and cell wall thickness of epidermal cells of the leaves, were detected. Finally, none plant-mediated UV effects neither in the whitefly B. tabaci nor in their parasitic wasp were found. Thirdly, the effects of a bifenthrin treated net were evaluated in different laboratory, semi-field and field experiments on the natural enemies studied. Treated nets were developed long time ago aiming at the control of the mosquitoes vectors of malaria, and nowadays, there is a great interest on assessing the possibility of their use in agriculture. In laboratory assays, a high mortality was recorded when E. mundus and A. swirskii adults were exposed by contact to the bifenthrin treated net for 72 hours in small cages (12 cm diameter). However, these natural enemies were not able to detect the presence of bifenthrin in a dual-choice test and no short-term mortality (72 hours) was recorded in those individuals that went through the treated net. In semi-field assays, performed under greenhouse conditions with cages of 25 x 25 x 60 cm high, the beneficial capacity of E. mundus was not affected. Finally, in field assays carried out in commercial multispan greenhouses (4000 m2) in Almería, A. swirskii was not affected by the presence of the bifenthrin treated net in the crop and the B. tabaci and F. occidentalis infestation levels were significantly lower than in the control. Finally, the composition of the microflora present in three species of parasitoids, E. mundus, Eretmocerus eremicus Rose & Zolnerowich and Encarsia formosa Gahan, and its influence in their susceptibility to insecticides, have been assessed. A total DNA extraction was performed on insects and universal bacterial primers were used to amplify the variable V4 region of the rRNA. A Next Generation sequencing (Illumina sequencing) was performed to identify the sequences of the bacterial genera present in the parasitic wasps. Once, the bacterial genera were identified, 16S rDNA gene of Actinobacteria were amplified from insects DNA extracts using the universal bacterial and actinobacterial primers, and the nested PCR products, were cloned to identify the Arthrobacter species. Three bacteria (A. aurescens Phillips, A. nicotinovarans Kodama, Yamamoto, Amano and Amichi and A. uratoxydans Stackebrandt, Fowler, Fiedler and Seiler), having the closest match with the Arthrobacter species present in the parasitic wasps, were obtained from the BCCMTM/LMG bacteria collection and its esterase activity was measured. Finally, antibiotic and residual contact tests were done to determine the influence of Arthrobacter species in the susceptibility of E. mundus to pesticides (abamectin). The results suggest that this bacterial genus can affect the toxicity of E. mundus to abamectin, which in turn supports the importance of the microbial community in natural enemies that it should be considered as a factor in risk assessment tests of pesticides.

Tuta absoluta (Meyrick, 1917) (Lepidoptera: Gelechiidae): adequação de uma dieta artificial e avaliação do seu controle biológico com Trichogramma pretiosum Riley em tomateiro

Tuta absoluta (Meyrick, 1917) é uma das pragas-chave da cultura do tomate e outras solanáceas na América do Sul e atualmente também na Eurásia e África. Devido aos grandes prejuízos que causa à cultura, são principalmente usados inseticidas para o seu controle. Entretanto, na busca de estratégias mais sustentáveis, cada dia adquire maior importância o uso do controle biológico, como uma das estratégias do manejo integrado de pragas. Para o desenvolvimento destas estratégias é fundamental desenvolver um método de criação de T. absoluta em laboratório, em dieta artificial, sem necessitar do hospedeiro natural, muitas vezes difícil de ser obtido e mantido em laboratório, e, de grande importância para produzir parasitoides específicos para esta praga. Dentre os parasitoides mais usados para ovos de lepidópteros está Trichogramma pretiosum Riley 1879 que é usado no controle biológico aplicado desta praga. Tendo como foco principal T. absoluta, neste trabalho foram pesquisados 1) a seleção de uma dieta artificial para este lepidóptero baseando-se em características físicas e químicas, avaliando o seu desempenho por várias gerações em laboratório, e 2) avaliação de aspectos biológicos e reprodutivos de T. pretiosum parasitando ovos de T. absoluta e aspectos físicos da planta (tricomas) para compreender o controle biológico desta praga no tomateiro. Foi encontrado que uma dieta à base de germe-de-trigo, caseína e celulose é apropriada para a criação deste lepidóptero, já que o inseto mostrou adaptação à mesma no transcorrer das gerações com base em características biológicas e de tabela de vida; adicionalmente, os ovos provenientes de T. absoluta alimentada com dieta artificial são comparáveis aos da dieta natural, no parasitismo de T. pretiosum. Com relação ao controle biológico foi demonstrado que este parasitoide desenvolvido em ovos de T. absoluta, diminui seu tamanho e desempenho com o transcorrer das gerações, apresentando menor capacidade de voo do que os insetos produzidos em A. kuenhiella, sendo necessária a liberação de altas densidades de parasitoides por ovo da praga. Foi observado que, embora o parasitismo de T. pretiosum de ovos de T. absoluta seja melhor em variedades com poucos tricomas, uma alta densidade destas estruturas não impede o controle da praga alvo dependendo da disposição destas estruturas. O controle biológico de T. absoluta com T. pretiosum tem uma ação momentânea, sendo necessárias liberações frequentes devido ao fato de os parasitoides desenvolvidos na praga serem menos competitivos com aqueles provenientes do hospedeiro alternativo que apresenta ovos maiores do que T. absoluta.

Evaluation of three application rates of Bacillus thuringiensis : efficacy and deposit analysis /

"April 1993"--P. [4] of cover.

Intra-specific variation for host plant use in Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae): Implications for management

The polyphagous moth Helicoverpa armigera (Hubner) is one of the world's most important agricultural pests. A number of existing approaches and future designs for management of H. armigera rely on the assumption that moths do not exhibit either genetically and/or non-genetically based variation for host plant utilization. We review recent empirical evidence demonstrating that both these forms of variation influence host plant use in this moth. The significance of this variation in H. armigera in relation to current and future pest management strategies is examined. We provide recommendations on future research needs and directions for sustainable management of H. armigera, under a framework that includes consideration of intra.-specific variation for host use relevant in this and other similar pest species. (C) 2004 Elsevier Ltd. All rights reserved.

Elicitors of induced disease resistance in postharvest horticultural crops: a brief review

Increasing loss of conventional fungicides due to pathogen resistance and general unacceptability in terms of public and environmental risk have favoured the introduction of integrated pest management (IPM) programmes. Induction of natural disease resistance (NDR) in harvested horticultural crops using physical, biological and/or chemical elicitors has received increasing attention over recent years, it being considered a preferred strategy for disease management. This article reviews the enhancement of constitutive and inducible antifungal compounds and suppression of postharvest diseases through using elicitors. The effect of timing of pre- and/or postharvest elicitor treatment and environment on the degree of elicitation and the potential for inducing local acquired resistance, systemic acquired resistance and/or induced systemic resistance to reduce postharvest disease is discussed. The review highlights that more applied and basic research is required to understand the role that induced NDR can play in achieving practical suppression of postharvest diseases as part of an IPM approach. (C) 2003 Elsevier B.V. All rights reserved.

Evaluation of the impact of natural enemies on Plutella xylostella L. (Lepidoptera: Yponomeutidae) populations on commercial Brassica farms

1 Accurate assessment of the impact of natural enemies on pest populations is fundamental to the design of robust integrated pest management programmes. In most situations, diseases, predators and parasitoids act contemporaneously on insect pest populations and the impact of individual natural enemies, or specific groups of natural enemies, is difficult to interpret. These problems are exacerbated in agro-ecosystems that are frequently disrupted by the application of insecticides. 2 A combination of life-table and natural enemy exclusion techniques was utilized to develop a method for the assessment of the impact of endemic natural enemies on Plutella xylostella populations on commercial Brassica farms. 3 At two of the experimental sites, natural enemies had no impact on P. xylostella survival, at two other sites, natural enemy impact was low but, at a fifth site, natural enemies drastically reduced the P. xylostella population. 4 The calculation of marginal death rates and associated k-values allowed the comparison of mortality factors between experimental sites, and indicated that larval disappearance was consistently the most important mortality factor, followed by egg disappearance, larval parasitism and pupal parasitism. The appropriateness of the methods and assumptions made to calculate the marginal death rates are discussed. 5 The technique represents a robust and easily repeatable method for the analysis of the activity of natural enemies of P. xylostella, which could be adapted for the study of other phytophagous pests.

Does the cutting of lucerne (Medicago sativa) encourage the movement of arthropod pests and predators into the adjacent crop?

Lucerne (Medicago sativa) has been suggested as an ideal refuge habitat as part of an integrated pest management (IPM) program because it harbours high numbers of beneficial arthropods. Whether or not cutting of lucerne encourages the movement of these beneficials into adjacent target crops is unknown. Vacuum samples were used to determine the effects of cutting lucerne on arthropod abundance (pests and predators) within lucerne and adjacent soybean (Glycine max) crops. Vacuum-sample collections of arthropods were conducted before and after lucerne cutting on seven occasions in four fields over two seasons. In the lucerne, 10 m by 1 m strips parallel to the crop interface were sampled at 5, 10, 15, 20 and 30 m from the interface. In the soybean, 10 m of row were sampled at the same distances from the crop interface. The abundance of predators in lucerne was reduced immediately after cutting at all distances from the interface. Predator abundance in soybean did not show any change. The cutting of lucerne significantly reduced pest numbers within the lucerne but had little effect on pest abundance in the adjacent soybean. The temporal pattern in pest and predator abundance was very different for each field sampled. Generally, arthropods decreased in abundance after cutting and gradually increased as the lucerne grew back. In soybeans, arthropod numbers fluctuated regardless of the cutting of the lucerne. Cutting of lucerne alone does not guarantee movement of predators into the adjacent target crop. The presence of lucerne fields within a cropping area may have some impact on regional predator populations, and so still be useful for IPM programs, but this has yet to be tested critically.

Phylogeography of the scarab beetle Antitrogus parvulus Britton (Coleoptera : Scarabaeidae) in south-eastern Queensland, Australia

This study surveys the population genetic structure of Childers canegrub, Antitrogus parvulus, to elucidate its population dynamics and gene flow. Antitrogus parvulus is a pest of sugarcane in the Bundaberg region and this knowledge can be used to optimise integrated pest management practices. Here, base-pair differences in the cytochrome oxidase II gene (COII) were used to characterise haplotypic diversity, infer levels of gene flow, and phylogenetic relationships of alleles and their phylogeographical structure. There were 28 unique haplotypes among the 70 sequenced individuals from the seven locations. All three variance components (among regions, among populations, within populations) are highly significant, with highest genetic diversity among regions and lowest among populations within regions. A positive correlation between migration rates and geographical distance and significant phylogeographical structure between four main geographical regions. The main implication of these findings for pest management is that if a grower can eliminate an existing infestation within a field, then reinvasion will be slow and further outbreaks within that field are unlikely to occur. The low dispersal ability of females also means that any resistance to insecticides that develops is likely to remain localised, but will rapidly become dominant within the affected population.

Temporal variation in arthropod sampling effectiveness: the case for using the beat sheet method in cotton

Predatory insects and spiders are key elements of integrated pest management (IPM) programmes in agricultural crops such as cotton. Management decisions in IPM programmes should to be based on a reliable and efficient method for counting both predators and pests. Knowledge of the temporal constraints that influence sampling is required because arthropod abundance estimates are likely to vary over a growing season and within a day. Few studies have adequately quantified this effect using the beat sheet, a potentially important sampling method. We compared the commonly used methods of suction and visual sampling to the beat sheet, with reference to an absolute cage clamp method for determining the abundance of various arthropod taxa over 5 weeks. There were significantly more entomophagous arthropods recorded using the beat sheet and cage clamp methods than by using suction or visual sampling, and these differences were more pronounced as the plants grew. In a second trial, relative estimates of entomophagous and phytophagous arthropod abundance were made using beat sheet samples collected over a day. Beat sheet estimates of the abundance of only eight of the 43 taxa examined were found to vary significantly over a day. Beat sheet sampling is recommended in further studies of arthropod abundance in cotton, but researchers and pest management advisors should bear in mind the time of season and time of day effects.

Facilitating IPM: The role of participatory workshops

Successful, long-term implementation of Integrated Pest Management (IPM) requires the integration of key technical and management activities and the participation of a wide range of stakeholders including farmers, researchers, extension officers, crop consultants, government agencies, and industry. A key issue that needs urgent attention is how to achieve the high quality interaction between these different groups which is necessary for sustained IPM. Problem specification and planning workshops (PSPWs) provide one means of facilitating an integrated strategy for tackling complex pest management issues. Since 1992, the Cooperative Research Centre for Tropical Pest Management has facilitated over 20 PSPWs, focusing on different farming systems in Australia. This paper describes the philosophy, the process involved, and the impact that these PSPWs have had. It examines three specific cases to describe the relationship between plans and results and ways of improving impact. The results reinforce the major role that social scientists can play in providing mechanisms for collaborating with technical researchers and other partners to facilitate effective, participatory ventures in IPM.

Delivering Regional Extension in Qld Farming Systems - Central Queensland

The CQ Cotton Regional Extension project has been a key to the delivery of emerging, cutting edge research information and knowledge to the Central Queensland cotton industry. The direct relevance of southern research to cotton production under the conditions experienced in CQ always has been an issue which could be addressed through regional assessment and adaptation. The project links the national research to the region through development and extension, with a strong focus on the major industry production issues including but not limited to disease, Integrated Pest Management (IPM), soils, nutrition and integrated weed management. Susan Mass has supported the implementation of national industry-wide programs particularly the industry Best Management Practices program (myBMP). This project has successfully transitioned to a focus on delivering national outcomes in target lead areas as part of National Development and Delivery Team established by Cotton CRC, CRDC and Cotton Australia, while maintaining a regional extension presence for Central Queensland cotton & grain farming systems. Susan Mass has very effectively merged and integrated strong regional extension support to cotton growers in Central Queensland with delivery of industry extension priorities across the entire industry in the Development and Delivery Team model. Susan is the target lead for disease and farm hygiene. Recognising the challenges of having regionally relevant research in Central Queensland, this project has facilitated locally based research including boll rot, Bt cotton resistance management, and mealybug biology through strong collaborations. This collaborative approach has included linkage to Department of Environment and Resource Managmeent (DERM) groups and myBMP programs resulting in a high uptake in CQ.

Estimating the demand curve for sustainable use of pesticides from contingent-valuation data

Stated-preference valuation techniques are often used to assess consumers' willingness-to-pay for food items produced in farming systems that adopt a sustainable use of pesticides (SUP). We propose an innovative valuation methodology in which dichotomous-choice contingent valuation is used to estimate the demand curve (price-quantity relationship) for such food items where price means price premium for the SUP output, quantity is the probability of choosing SUP and the conventional food product is kept available in the market at the current market price. This methodology can be used to evaluate market differentiation as a policy option to promote the SUP. The methodology is tested with data from a sample of urban consumers of fruits and vegetables in Portugal. The estimated demand curve is used to define the price level maximizing the total premium revenue for the SUP sector as a whole. This optimal level of the price premium is €77.55 (or 163% of the value of the monthly basket of fruits and vegetables at current prices). Adopting the optimal price premium will decrease the number of consumers of SUP food by 54%. The reduction is even higher for low income consumers (80%) leaving them more exposed to the risks of pesticide use.