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1 IOBC-WPRS Bulletin Vol. 80, 2012 Working Group Integrated Control in Protected Crops, Mediterranean Climate. Preceedings of the meeting at Catania, Sicily (Italy), October, Edited by: Cristina Castañé, Rosa Gabarra, Judit Arnó, Luciana Tavella and Giovanna Tropea-Garzia. ISBN [VIII+ 344 pp.]. Preface... I Contents... III Key notes Prospects and challenges for biological pest control in protected crops in the Mediterranean Karel Bolckmans Abstract: The Mediterranean is a major producer of protected crops. Especially during the past six years the uptake of biological control of pests has increased rapidly in the Mediterranean. The key challenges for growers are pesticide resistance management, residue management, regulatory requirements, risk management and cost-effectiveness. Suggestions are provided for continued research on improving biological control. Biological and integrated protection in the Mediterranean greenhouse: is disease management the weak link? Philippe C. Nicot, Marc Bardin Abstract: While biocontrol is commonly used for the protection of greenhouse crops against arthropods, continued reliance on chemical control against diseases is often considered as a hindrance to integrated protection. This paper presents a review of currently available biocontrol products and other alternatives to pesticides for use against plant diseases in the Mediterranean greenhouse. At least 12 biological active substances (micro-organism or plant extract) are commercially available for the control of soilborne pathogens and 14 against foliar diseases, in one country or more in the Mediterranean Basin. Future prospects and challenges are presented. Natural antagonists and insect control: a new perspective Francesco Pennacchio Abstract: The astonishing biodiversity of parasitic Hymenoptera and the large variety of strategies they use to colonize and exploit their insect hosts offer a wealth of molecular tools that can be profitably used for pest control. The virulence factors encoded by parasitic wasps and their associated symbionts are able to suppress the immune response of the host and to disrupt its physiological and reproductive processes. These molecules can directly target insect pests, and, by disrupting their immune response, they can also enhance the value of the ecological services offered by natural antagonists. The use of these natural bionsecticides is highly dependent on the development of targeted delivery strategies, which have to be designed to enhance both the efficacy and safety. The fast expansion of molecular technologies opens new avenues of research and makes possible the development of innovative pest control technologies, based on the use of parasitoid-derived molecules and/or on strategies mimicking their effects on insect hosts. Integrating community ecology theories for optimizing IPM Nicolas Desneux III

2 IV Pest dynamics and surveys of their native natural enemies Survey of host plants and natural enemies of Drosophila suzukii in an area of strawberry production in Catalonia (northeast Spain) Judi Arnó, Jordi Riudavets, Rosa Gabarra Abstract: A new invasive pest, the fly Drosophila suzukii, is threatening several fruit crops in Europe. This pest is native to Asia and was first recorded in Europe in The first documented damage in our area occurred in 2011, mainly affecting cherries and strawberries. In this study, we aimed to determine which cultivated and non-cultivated plants host D. suzukii in an area in which heavy infestations on protected strawberry crops have occurred. Strawberries, raspberries, Arbutus unedo and Solanum luteum have been infested with D. suzukii. To our knowledge, the last two species had not previously been described as hosts of this fly. Evidences of the presence of the pest in unripe fruits were recorded. Some potential biological control agents in the local area were also identified. Four Hemiptera predator species were found in infested fruit samples: Orius laevigatus, Cardiastethus nazarenus, C. fasciventris and Dicyphus tamaninii. Further research is needed in the role of these natural enemies in the control of this pest. The cotton bollworm Helicoverpa armigera (Lepidoptera: Noctuidae) is it becoming a serious pest on pepper and tomato in Montenegro? Sanja Radonjić, Snježana Hrnčić Abstract: In the last few years, increases in the damage to pepper and tomato fruits caused by the cotton bollworm Helicoverpa armigera have been noticed in the southern part of Montenegro, the country s main pepper and tomato production area. This includes the area around the city of Podgorica (Zeta and Bjelopavlići) and the Montenegro seacoast (Ulcinj) where these two cultures are grown both in greenhouses and outdoors. From July to September in 2010 and 2011, eight tomato and six pepper greenhouses in the Zeta and Bjelopavlići area were visually inspected. Two tomato greenhouses, one open-field tomato crop and one open-field pepper crop were visually inspected in 2011 in the Ulcinj area. As well as a visual inspection, the intensity of attack was also calculated for the pepper and tomato crops. The results of this study showed that larvae of H. armigera were found on pepper and tomato fruits in most of the inspected greenhouses and also on tomato fruits in the open field. More damaged pepper and tomato fruits were observed in 2011 than in A three-year survey of Tuta absoluta (Lepidoptera: Gelechiidae) population trends in Sardinian tomato greenhouses Mauro Nannini, Fabrizio Atzori, Michele Coinu, Riccardo Pisci, Francesco Sanna Abstract: Since 2008 Tuta absoluta has established in Sardinia (Italy) as the key pest of greenhouse tomatoes. With the aim of monitoring tomato borer population trends after its introduction in one of the most important fresh market tomato production areas of the island, from February 2009 to January 2012 we surveyed ten to fifteen commercial greenhouse crops per year. In each crop we assessed monthly the mean number of live T. absoluta larvae per tomato plant, the percentage of plants infested by larvae, the mortality and parasitism of 2 nd -4 th -instar larvae, and noted the treatment schedules. The highest levels of tomato borer infestation were observed in spring of 2009 (averaging 86 larvae/plant) and, to a lesser extent, in spring of 2010 and 2011 (55 and 41 larvae/plant, respectively). While pest density generally declined in summer, it increased moderately during autumn, not exceeding on average 8 larvae/plant, and was maintained in winter. In most cases the highest levels of larval mortality were recorded between May and July. Except in a few cases, the contribution of larval parasitism to tomato borer mortality was poor. The insecticides most commonly used by growers for pest management were initially abamectin, azadirachtin and spinosad, but during the survey period the latter two products were gradually replaced by Bacillus thuringiensis-based insecticides and emamectin benzoate.

3 V Tomato leafminer, Tuta absoluta, and insecticide resistance: a new challenge for control strategies Khalid Haddi, Madeleine Berger, Pablo Bielza, Dina Cifuentes, Linda M. Field, Kevin Gorman, Martin S. Williamson, Chris Bass, Carmelo Rapisarda Abstract: The tomato leaf miner, Tuta absoluta (Lepidoptera), is a significant pest of tomatoes that has undergone a rapid expansion across Europe, North Africa and parts of Asia during the past six years. One of the main means of controlling this pest is through the use of chemical insecticides. In the current study leaf-dip bioassays were used to determine the susceptibility of T. absoluta strains established from field collections to several insecticides. Additionally, the para-type sodium channel and acetylcholinestaerase gene from T. absoluta were cloned and sequenced. This revealed the presence of three kdr/super-kdr-type mutations (M918T, T929I and L1014F) in the sodium channel gene and mutation (A201S) in the acetylcholinesterase gene. To assess the prevalence of these mutations in 27 field strains from 12 countries high-throughput TaqMan diagnostic assays were developed. The results showed the presence of these mutations at high frequency in T. absoluta populations world-wide and suggests pyrethroids and organophosphates are likely to be ineffective for control. These results also support the idea that the rapid expansion of this species over the last six years may be in part mediated by the resistance of this insect to chemical insecticides hence adding new challenges to control strategies for this pest. Promising native candidates for biological control of Tuta absoluta in Italy Chiara Ferracini, Barbara L. Ingegno, Marco Mosti, Paolo Navone, Luciana Tavella, Alberto Alma Abstract: Tuta absoluta (Lepidoptera: Gelechiidae), native to central America, has been first reported in Europe in Spain in 2006 and in southern Italy in 2008, and by now it has become one of the major pests on tomato, its main host plant. In the two-year period , nine species of indigenous parasitoids emerged from tomato leaves infested by T. absoluta collected in horticultural areas of Liguria, Sardinia and Sicily (Italy). The most abundant species were Necremnus near artynes and N. near tidius, which appeared to be promising as biological control agent. Furthermore, in tomato plantations infested by T. absoluta in Piedmont (Italy), a generalist predator, Dicyphus errans (Heteroptera: Miridae), was largely found. Therefore, the two parasitoids and the mirid bug were tested in controlled conditions to evaluate their effectiveness in controlling the exotic pest. Both Necremnus species proved to control efficiently first- and second-instar larvae by host feeding and parasitism. Also D. errans revealed to be an effective biocontrol agent of the tomato borer, preying mainly on eggs and first-instar larvae. Further investigations are needed to evaluate their potential role as biological control agents in commercial tomato plantations. Survey on indigenous parasitoids of the invasive exotic pest Tuta absoluta in Southern Italy Lucia Zappalà, Umberto Bernardo, Antonio Biondi, Arturo Cocco, Salvatore Deliperi, Gavino Delrio, Massimo Giorgini, Paolo A. Pedata, Carmelo Rapisarda, Giovanna Tropea Garzia, Gaetano Siscaro Abstract: The tomato borer, Tuta absoluta, is an invasive pest native to South-America and since its arrival in Europe the tomato production has faced severe yield loss. The composition of the indigenous parasitoid complex of the leafminer was monitored in Southern Italy (Campania, Sardinia and Sicily) during The parasitoid collection was carried out by exposing sentinel tomato infested plants and by sampling open field and protected greenhouse crops, as well as Solanum nigrum, a T. absoluta wild host. The parasitoids found developing on the leafminer were mostly generalist idiobionts belonging to 12 genera and 6 families (Ichneumonidae, Braconidae, Eulophidae, Elasmidae, Pteromalidae and Trichogrammatidae). A prompt shift of native parasitoids to the new invasive host was observed and the parasitoid complex recovered on T. absoluta seems to follow the typical pattern of parasitisation on exotic pests, being characterized by a relatively low number of species mostly represented by generalist idiobionts, performing low levels of parasitisation in open field. This study highlighted the

4 VI suitability of sentinel plants for indigenous parasitoids surveys in case of heavily treated crops, since the majority of the species were collected on sentinel plants (16 out of 23 totally recovered). The data recorded up to now lead to be rather optimistic with reference to the perspective of T. absoluta natural biocontrol in the Mediterranean basin. Can recently found Brazilian hemipteran predatory bugs control Tuta absoluta? Vanda H. P. Bueno, Flavio C. Montes, Ana M. C. Pereira, Juracy C. Lins Jr., Joop C. van Lenteren Abstract: The tomato borer Tuta absoluta, native to western South America, is an extremely devastating pest in tomato crops in most of South America, Europe and Africa North of the Sahel and the Near East (Asia). Without control, the pest causes yield losses up to 100% and decreases fruit quality in open field and greenhouse crops. In Brazil two other serious lepidopteran pests occur in tomato, as well as thrips, whitefly, mites and aphids. For control of these pests, frequent applications of pesticides of up to 5 times per week are needed, and these resulted in the appearance of resistant populations to a number of active ingredients and caused a strong reduction of natural enemies. Biological control may offer a better opportunity for pest management. Mirid predatory bugs are currently used with success in Europe to control T. absoluta and other pests. In Brazil, three mirid predators of T. absoluta have been found and are now evaluated together with two other hemipteran predators, Geocoris punctipes and Orius insidiosus. Natural parasitism of Bemisia tabaci (Hemiptera: Aleyrodidae) by native Aphelinidae (Hymenoptera) parasitoids in tomato greenhouses in Mersin, Turkey Kamil Karut, Cengiz Kazak, İsmail Döker, Amir Abdullahi Yousif Malik Abstract: The survey study was conducted to determine the natural parasitism of Bemisia tabaci in tomato greenhouses in the Adanalıoğlu, Kazanlı and Kocahasanlı counties of Mersin in the period Samples were collected from commercial tomato greenhouses in the spring and autumn seasons. Most of the surveyed greenhouses ( 95%) were infested with B. tabaci. Overall, population densities of B. tabaci were higher in Adanalıoğlu and Kazanlı than in Kocahasanlı. B. tabaci infestation rates were higher in autumn than in spring, in both years. Er. mundus was more abundant in autumn than in spring at all locations and Er. mundus was more abundant than En. lutea. Parasitism rates of Er. mundus ranged from 12.3 to 37.1% throughout the survey. Detection of Tuta absoluta (Lepidoptera: Gelechiidae) in Iran Ahmad Cheraghian Implementation of a biological control programme in greenhouse crops in Iran Vali Baniameri, Ahmad R. Mohandessi, Shahram Farrokhi Basic biological characteristics of natural enemies Morphometry and biology of a new anthocorid Montandoniola indica, a potential predator of Gynaikothrips uzeli Chandish R. Ballal, Tripti Gupta, Sunil Joshi Abstract: A new anthocorid predator, Montandoniola indica was recorded for the first time as a predator of Gynaikothrips uzeli infesting Ficus retusa in Karnataka (India). The morphometry and biology of this predator were studied. M. indica nymphs and adults could be reared on UV irradiated eggs of the rice moth Corcyra cephalonica. The feeding potential of M. indica nymph was 27 C. cephalonica eggs and in the case of adult male and female, 38 and 56 eggs,

5 VII respectively. The adults of M. indica were provided with thrips and pollen in addition to C. cephalonica eggs, for better survival. The adult male and female longevity was 23 and 31 days, respectively and fecundity 37 eggs per female. Morphometry and molecular markers to discriminate between Macrolophus pygmaeus and Macrolophus melanotoma Cristina Castañé, Núria Agustí, Judit Arnó, Rosa Gabarra, Jordi Riudavets, Jordi Comas, Óscar Alomar Abstract: Macrolophus melanotoma (= M. caliginosus) and Macrolophus pygmaeus have been referred to as important predators of pests in vegetable crops in the Palaearctic region. Due to their high morphological similarity, these two mirid species have been confused, with important consequences for their use in inoculative and/or conservative biological control programmes. In this study we review the taxonomic characters that proved reliable to separate the two species, as the profiles of cuticular hydrocarbons, a discriminant function of four morphometric ratios of males, and specific primers to distinguish both species through conventional PCR. We also discuss the correct assignation of Macrolophus species in historical records. All the tested Macrolophus samples from the IRTA s old lab colonies were identified as M. pygmaeus, showing that the articles we have previously published on M. caliginosus using laboratory colonies should have referred to M. pygmaeus. When samples from comercial companies were analyzed using the previously mentioned specific primers, in all cases, the species was identified as M. pygmaeus. This result shows that in most cases the species being commercially released is M. pygmaeus. We can therefore hypothesize that most of the field work done on M. caliginosus in the Mediterranean Basin, and presumably also in the rest of Europe, which has mainly focused on tomato crops, could probably have referred to M. pygmaeus. The identification of native plants that are sources of the predator that colonises the tomato crop is a key element in the implementation of conservation biological control programmes for this crop. However, most of the host plants for both species still remain largely unknown. Influence of phytophagous behaviour on prey consumption by Macrolophus pygmaeus D. Maselou, D. Perdikis A. Fantinou Abstract: Omnivorous Heteroptera constitute an important component of predatory guilds with high potential for biological control. Understanding the relative effects of plant feeding on the suppression of prey by omnivores could be an important element for improving biological control strategies. In the current paper, the effects of different plant food sources on the predation rate of the omnivorous predator Macrolophus pygmaeus (Hemiptera: Miridae) were examined. In all the experiments, second instar nymphs of the aphid Myzus persicae (Homoptera: Aphididae) were used as prey at different densities. First, we evaluated the rate at which the predator preyed on M. persicae at various prey densities on pepper and eggplant leaves. Then, using eggplant flowers or pollen as additional food sources, we estimated predator efficiency for three different prey densities. The predation rate was not affected by the type of plant leaf used. However, the results showed that the predation rate of M. pygmaeus was significantly reduced when flowers or pollen were provided at high prey densities. The importance of these results in understanding the influence of phytophagy on predation rates of omnivorous predators is discussed. Effect of supplemental food on the fitness of four omnivorous predator species Marta F. Oveja, Judit Arnó, Rosa Gabarra Abstract: The predators Orius laevigatus, O. majusculus, Macrolophus pygmaeus and Nesidiocoris tenuis are extensively used in biological controls in vegetable crops. The fitness and local density of these predators, which are omnivores and feed on arthropods and plants, may be enhanced by the provision of supplemental food, especially when prey is scarce, and this may improve biological control results. The aim of our study was to evaluate the effect of four supplemental foods: Ephestia kuehniella eggs, dry Artemia sp. cysts, the mite Tyrophagus putrescentiae and commercial multifloral bee pollen, on the reproduction and longevity of these

6 VIII four predators. Our results show that using dry cysts of Artemia sp. as factitious prey had positive effects on the reproduction of the predators O. majusculus, O. laevigatus, M. pygmaeus and N. tenuis. Furthermore, dry commercial multifloral bee pollen improved the longevity of the females of O. laevigatus, O. majusculus and M. pygmaeus as much as eggs of E. kuehniella. Dry cysts and dry pollen need little or no handling to prepare them for use, are very easy to disperse in the crop and maintain their quality for a reasonable time. Further experiments need to be conducted under semi-field conditions to better evaluate the positive effect of these two supplemental foods on predator establishment in the crop. It will also be important to evaluate the effect of these foods on life parameters of pest species and on intraguild relationships among predators. Analysing predation of Orius majusculus (Heteroptera: Anthocoridae) in lettuce crops by PCR Priscila Gómez-Polo, Oscar Alomar, Cristina Castañé, Nuria Agustí Abstract: Orius majusculus (Hemiptera: Anthocoridae) is an important generalist predator known to feed on two of the main pests in lettuce crops, Frankliniella occidentalis (Thysanoptera: Thripidae) and Nasonovia ribisnigri (Hemiptera: Aphididae). Diagnostic molecular gut-content analysis has been conducted to study predation of O. majusculus on these pests, as well as on the springtails of the genus Entomobrya sp. (Collembola), the most abundant non-pest prey in lettuce crops of the studied area. Two pairs of specific primers were designed for N. ribisnigri and F. occidentalis, amplifying 331 bp and 277 bp fragments of the mitochondrial Cytochrome c Oxidase I (COI) region, respectively. Collembola detection, was conducted with a pair of a group-specific primers previously designed that amplify an amplicon of 177 bp of the 18S rdna region. Prey was detected in 100% of predators immediately after prey consumption (t = 0h). PCR analysis of O. majusculus collected in the field revealed a strong relationship between prey density in the crop and consumption. Sixty per cent of these field-collected O. majusculus were positive for N. ribisnigri in spring (when this pest species was present) and up to 40% were positive for F. occidentalis in summer (when the density of this pest was the highest). Collembolan detection ranged from 9.4% to 28.2% depending on the season and predator stage. A higher proportion of immature O. majusculus ingested more than one prey species simultaneously compared to adults of the same predator species. These findings suggest that O. majusculus is an important predator for the control of both lettuce pests, being springtails an important alternative prey, which could maintain O. majusculus populations when both pest species are not present in the crop or are not abundant enough. The effect of pollen on the predation of Polyphagotarsonemus latus by the predaceous mites Amblyseius swirskii and Euscelis scutalis Huda Al-Beiruti, Yael Lubin, Phyllis G.Weintraub Do oviposition substrates and different materials as hiding-places influence reproductive parameters of Orius insidiosus? Flávio C. Montes, Vanda H. P. Bueno, Juracy C. Lins Jr, Adriana A. Menezes, Ana M. Calixto, Nazaré Moura Abstract: Orius insidiosus is a commercially produced natural enemy used as a biological control agent of thrips in greenhouses. Mass production is strongly influenced by the type of oviposition substrate, as well as by hiding materials that reduce the contact between individuals and avoid cannibalism in the rearing unit. The objective of this study was to evaluate two oviposition substrates, bean sprouts and bean pods of Phaseolus vulgaris, and different types of materials (rice hulls, popcorn, shredded paper towel and Styrofoam white flakes) to provide hiding places and prevent cannibalism in the rearing system of O. insidiosus. Bean sprouts were accepted and more suitable for O. insidiosus females as an oviposition substrate in the laboratory. The total number of eggs/rearing unit/30days, the number of eggs/female/day/rearing unit and the number of emerging adults were significantly higher with bean sprouts as oviposition substrate

7 IX compared to bean pods. No significant differences were found in the number of eggs and number of emerging adults/rearing unit with different types of materials to provide hiding-places and avoid cannibalism. Do earthworms increase oviposition of the tiger-fly, Coenosia attenuata (Diptera: Muscidae)? Elisabete Figueiredo, Susana Leal, Joana Martins, Célia Mateus, Teresa Rebelo Prevention of diapause induction in a Japanese strain of Aphidoletes aphidimyza Eizi Yano, Masafumi Yamane Abstract: The aphidophagous gall midge Aphidoletes aphidimyza (Diptera: Cecidomyiidae), a dominant natural enemy of aphids, is used in many countries to control aphids in greenhouses. Since developmental arrest during diapause reduces the effectiveness of natural enemies, we studied the effect of photoperiod and temperature on the incidence of diapause in a Japanese strain of A. aphidimyza by examining diapause induction under different day-length and temperature conditions. The critical day length for diapause induction was 12.7h at 20 C. Diapause incidence was completely prevented at 30 C, even under a photoperiod of 11L13D. Diapause induction in A. aphidimyza could be prevented by providing light during the scotophase in the laboratory. However, diapause induction was not prevented by providing light for 1h at night in the field experiments from September to November A comparison of three Encarsia species as controlling agents for Bemisia tabaci in greenhouses Dan Gerling, Nelly Rejouan Abstract: The study constitutes part of an effort to locate potentially efficient agents for biological control against the whitefly Bemisia tabaci. Three parasitoid species, Encarsia inaron, E. lutea and E. sophia, which attack B. tabaci under natural conditions were studied comparatively in order to evaluate the most suitable species for use as a biological control agent against Bemisia tabaci in greenhouses. The tested parameters were: developmental durations, longevity and oviposition characteristics. All three species lived ca. 20 days each and the two former ones developed to adulthood within ca. 14 days and E. lutea, within 15. The differences between oviposition rates were not significant (66 ± 36.27, ± and ± average and standard deviation of the mean respectively for E. inaron, E. lutea and E. sophia respectively). Encarsia inaron had the highest percentage of females that lived longest whereas E. lutea had a higher semelparity. Our findings failed to reveal the superiority of one species over the others as candidates for the intended biological control. Additional parameters, including their competitive behavior and compatibility with other natural enemies in the greenhouse, should thus be considered. Biology and behaviour of the indegenous parasitoid Bracon nigricans on the invasive South American tomato pinworm Tuta absoluta Antonio Biondi, Gaetano Siscaro, Nicolas Desneux, Edwige Amiens-Desneux, Lucia Zappalà Dispersal of TSWV by Frankliniella occidentalis (Thysanoptera: Thripidae) in presence of natural enemies Belén Belliure, Juan Antonio López-Adánez, Alberto Fereres, M. Ángeles Marcos-García Extended Abstract

8 X Biology of natural enemies at greenhouse scale Effects of selected factors on the reproductive fitness of the predatory mirids Macrolophus pygmaeus and Nesidiocoris tenuis (Heteroptera: Miridae) Mauro Nannini, Michele Coinu, Giovanni Murgia, Riccardo Pisci, Francesco Sanna Abstract: The use of the predatory mirids Macrolophus pygmaeus and Nesidiocoris tenuis for the control of arthropod pests on greenhouse tomatoes is at present limited in some Mediterranean regions by factors of an economic and technical nature. To assess the influence of the production and distribution process, the availability of a supplementary food source and seasonal climatic trends in the reproductive fitness of the two mirid species, some laboratory and semi-field experiments were conducted during the last year. The adverse effects of the stress experienced by commercially supplied insects have clearly emerged through the investigation, but at different levels of intensity for the sources tested. The supply of Ephestia kuehniella eggs has shown pronounced effects on the reproductive fitness of the predatory mirids, in some cases allowing a partial recovery of fertility in stressed individuals and confirming the suitability of supplementary food sources in improving mirid performances. Furthermore, the results of the tests carried out appear to indicate that M. pygmaeus is more suitable for winter releases in Sardinian tomato greenhouses, whereas N. tenuis is to be preferred for autumn releases. Use of the predators Orius laevigatus and Aeolothrips spp. to control Frankliniella occidentalis populations in greenhouse peppers in the region of Monastir, Tunisia Mohamed Elimem, Brahim Chermiti Abstract: The use of Oriuslaevigatus and Aeolothrips spp. to control Frankliniella occidentalis in greenhouse peppers in Tunisia produced different results according to predator species and different doses and release frequencies. In the case of the predator bug Orius laevigatus, the most effective dose was 1 individual per m² repeated three times with an interval of one week. Although O. laevigatus releases caused F. occidentalis populations to decrease for only one week, the predator was able to become installed in the crop and to proliferate. In the case of the predatory thrips Aeolothrips spp., no individuals were recorded in the pepper greenhouse even after the third release. However, the F. occidentalis population decreased to low average values of 0.42 and 0.06 thrips per flower as a result of the spontaneous colonization of individuals of O. laevigatus from other greenhouses. Effect of Orius laevigatus and Amblyseius swirskii releases on Frankliniella occidentalis populations in pepper crop greenhouses in the Bekalta region of Tunisia Mohamed Elimem, Brahim Chermiti Evaluation of four lacewing species for aphid control in sweet pepper Gerben Messelink, Chantal Bloemhard, Hans Hoogerbrugge, Jeroen van Schelt Abstract: Four species of lacewings were evaluated for control of the peach aphid Myzus persicae in sweet pepper. Chrysopa perla was very effective in controlling aphids, probably because the adults also consume aphids. All other lacewings did not reduce aphids numbers sufficiently. Chrysoperla lucasina was establishing better than Chrysoperla affinis. The hemerobiid Micromus variegatus seems to prefer the lower plant parts. Since this species is relatively small, more individuals are needed to control aphids. Preliminary releases in large greenhouses suggest that none of the lacewing species establish well when released as adults. Further research is needed to trigger them to stay and oviposit into a sweet pepper crop.

9 XI Improved biological control of problem aphids on protected herbs Jude Bennison, Tom Pope, Joanna Greetham, Tracie Evans, Heather Maher Abstract: Hawthorn-parsley aphid, Dysaphis apiifolia and mint aphid, Ovatus crataegarius are problem aphid species on protected parsley and mint respectively. Grower experience has indicated that these aphids are not parasitised by the three aphid parasitoid species available until recently, i.e. Aphidius colemani, Aphidius ervi and Aphelinus abdominalis. A new mix of six parasitoid species is now available, including the above three species and in addition, Aphidius matricariae, Ephedrus cerasicola and Praon volucre. In laboratory bioassays, all six species except for A. ervi parasitised hawthorn-parsley aphid, and A. matricariae, E. cerasicola and P. volucre parasitised mint aphid. In semi-field glasshouse cage experiments, A. colemani and A. matricariae parasitised and reduced numbers of hawthorn-parsley and mint aphids respectively on parsley and mint. Experimental studies of the biological control of aphids in protected strawberry crops in France Yannie Trottin-Caudal, Véronique Baffert, Jean-Michel Leyre Abstract: Integrated pest management is increasingly used by growers in France in protected strawberry crops due to difficulties with chemical control, especially on soilless crops. Various different pests can attack strawberries, including: aphids, mites, thrips, tarsonemids, mirid bugs and, more recently, Drosophila suzukii. A survey was carried out from 2010 to 2012 in the southeast of France that confirmed the presence of up to six aphid species. Since 2008, promising trial results from aphid control have been obtained in experimental tunnels, based on the efficiency of lacewings that were released onto aphid-infested plants and parasitism by certain parasitoids adapted to aphid species. Possibilities for integrated control of citrus mealybug in commercial ornamental greenhouses Juliette Pijnakker, Ada Leman Abstract: Since the 2000 s, the citrus mealybug Planococcus citri has become a key pest in integrated ornamental greenhouse crops in The Netherlands. While adopting a more integrated approach toward controlling other pests, growers have abandoned regular applications of broad spectrum pesticides, thereby allowing mealybugs to become more widespread. The benefits of both inundative and inoculative releases of (commercially available) natural enemies of this pest have been studied. Three encyrtid parasitoid species: Coccidoxenoides perminutus, Leptomastix dactylopii and Anagyrus pseudococci were tested on potted plants Epipremnum aureum infested with citrus mealybug Planococcus citri. Parasitoid performance was evaluated under confined conditions in large cages. Weekly release of L. dactylopii and A. pseudococci supressed the development of small hot spots and prevented the spread of mealybug from infested plants to healthy plants, but did not result in pest eradication. Most Epipremnum plants treated by these two parasitoids were marketable. C. perminutus gave insufficient control. The relevance of introducing natural enemies for controlling mealybugs in greenhouse ornamentals is discussed. Selection of Trichogramma spp. (Hym.: Trichogrammatidae) for the biological control of Tuta absoluta (Lep.: Gelechiidae) in greenhouses by an entomo-ecological simulation model T. Cabello, M. Gámez, Z. Varga, J. Garay, R. Carreño, J. R. Gallego, F. J. Fernández, E. Vila Abstract: A greenhouse trial was carried out, between October 2011 and April 2012, on tomato crops with the aim of evaluating the biotic potential of two species of Trichogramma (T. achaeae and T. urquijoi) for the control of the South American tomato moth, Tuta absoluta. The trial also studied the relationship of the latter with the predator Nesidiocoris tenuis, which is also used in biological control programmes in greenhouses. With this objective, we developed a mathematical model which makes it possible to evaluate and compare the effects of different biological control agents. As a result we established that T. achaeae was better at controlling pest populations than

10 XII any other species. We also noted a significant intra-guild competition effect of the predator on both species of Trichogramma. Biological control of Tuta absoluta (Lepidoptera: Gelechiidae) in protected tomato crops in Argentina M. Gabriela Luna, Patricia P. Pereyra, Norma E. Sánchez Abstract: We present results of a ten-year research project on life history traits and field interaction of two main Tuta absoluta indigenous larval parasitoids, Pseudapantales dignus and Dineulophus phtorimaeae. These natural enemies naturally coexist in cropping conditions, show positive characteristics as biological control agents against T. absoluta by means of augmentative releases in argentinean greenhouse tomato. Releasing syrphid larvae (Diptera: Syrphidae) as an effective aphid biocontrol strategy in Mediterranean sweet-pepper greenhouses Rocco Amorós-Jiménez, Belén Belliure, Lucía González Franco, Águeda Pose-Pardiñas, Alberto Fereres, M. Ángeles Marcos-García Multi-stage dynamic model for prey-predator interaction: Application to Spodoptera exigua (Lep.: Noctuidae) and Nabis pseudoferus (Hem.: Nabidae) under greenhouse conditions M. Gámez, Z. Sebestyén, Z. Varga, J. Garay, J. R. Gallego, F. J. Fernández, T. Cabello Side effects of pesticides on natural enemies Insecticides and beneficial predators: side effects on Orius spp. on IPM pepper and strawberries Lara Bosco, Nicola Bodino, Michele Baudino, Luciana Tavella Abstract: The species of the genus Orius (Heteroptera: Anthocoridae) are well-known as generalist predators able to control thrips outbreaks on different crops, such as pepper and strawberries. The assessment of the compatibility of various insecticides with predatory activity is a key for the success of IPM strategies, including Orius releases on crops. During 2011, experiments were carried out in Piedmont (northwestern Italy) on four commercial pepper and strawberry tunnels. Toxicity of the most used insecticides were also evaluated in laboratory bioassays on wild Orius spp. Abamectin, λ-cyhalothrin, and acrinahtrin proved to be not compatible with these predators, both in field and laboratory experiments. Azadirachtin was almost harmless for Orius spp. whereas spinosad induced high levels of mortality in laboratory bioassays but seemed to be safe in field experiments. Etofenprox gave contradictory results in the field and laboratory experiments, corroborating the need of multiple testing methods in evaluating the effects of pesticides on beneficial insects. Chlorantraniliprole (Rynaxypyr, Coragen, Altacor ) key features for sustainable control of Tuta absoluta Andrea Bassi, Jean Luc Rison, Emmanouil Roditakis, Luigi Sannino Abstract: Chlorantraniliprole (Rynaxypyr ) is a novel diamide insecticide by DuPont with outstanding performance on Tuta absoluta and an extremely low mammalian toxicity profile. Tested on T. absoluta since 2002 in Brasil at the DuPont R&D Station in Paulinia, early results indicated a new standard of T. absoluta control, even on insecticide-resistant populations. Since 2007 numerous field and laboratory studies have been carried out in the Mediterranean region by DuPont and independent researchers. These studies have contributed to in-depth knowledge of its

11 XIII features and to finalize the recommendations for use in a European IPM context. This review provides a selection of field and laboratory observations relative to the product selectivity to key natural enemies and the baseline sensitivity of T. absoluta populations from the Mediterranean countries and the product best use strategy for IRM (Insecticide Resistance Management). Testing for non-target effects of some fungicides and insecticides on western flower thrips and their predator Amblyseius swirskii under plastic tunnel conditions Halil Kutuk, Mehmet Karacoglu Abstract: The compatibility of four pesticides (Chlorantraniliprole, Pymetrozine, Trifloxystrobin and Myclobutanil) used with the predatory mite, Amblyseius swirskii (Acari: Phytoseiidae), to control western flower thrips, Frankliniella occidentalis (Thysanoptera: Thripidae), was investigated on peppers in plastic tunnels. One preventative release of predatory mites, at the rate of 50 adults per m 2, was made from the stock culture when the peppers started to flower. The western flower thrips populations were sampled on a weekly basis and the tested pesticides were sprayed when needed. Our results indicated that applying the tested pesticides had no effect on the ability of the predator to reduce the thrips population. The thrips populations in the nopredator treatment continued to expand throughout the experiment, while those in the predator release treatment declined. We concluded that A. swirskii can be used in conjunction with the tested pesticides in pepper greenhouses without causing any obvious detrimental effects to this predator or reducing the effectiveness of the biological control. Biological control of the spider mite Tetranychus urticae and interaction between beneficial organisms on vegetable crops in greenhouses Żaneta Fiedler, Danuta Sosnowska Abstract: Tetranychus urticae is the most important pest on vegetable crops in greenhouses in Poland and successful control of this pest is very difficult. The present registration requirement for biological control agents in Poland states that macro-organisms such as nematodes and beneficial organisms do not need to be registered. As a result, a lot of new natural enemies are now commercially available and nine species can be used against spider mites. Research work was conducted on the interaction between natural enemies introduced into greenhouse-grown tomato crops to reduce the number of pests such as spider mites and the side effects of insecticides and fungicides on predators. This interaction was studied with reference to the predatory mite species: Amblyseius swirskii and Phytoseiulus persimilis T which are used in laboratory and greenhouse tests to control the two-spotted spider mite Tetranychus urticae. The greatest efficacy (86%) in limiting the number of T. urticae was obtained through the use of P. persimilis and A. swirskii. The results of this experimental treatment were statistically different from other treatments involving both the separate and combined use of specific predatory mite species. The study also revealed competition between the two species A. swirskii and P. persimilis, with the predatory mite A. swirskii being the dominant species. The insecticides Abarex, Spin Tor and Nissorun were safe for use with predatory mites and could be recommended for IPM (Integrated Pest Management) programmes for greenhousegrown crops. The result of the studies carried out revealed that the species P. persimilis was the most susceptible to the different fungicides used, based on laboratory experiments, and especially when the predator was released 1 and 3 days after applying these fungicide treatments. In general, the fungicides applied showed a lower level of toxicity to all the beneficial organisms when the predatory mites were released 5 days after its application. The fungicide Topsin 500 SC appeared to be selective with respect to the species A. swirskii. Side effects of bioinsecticides used to control Tuta absoluta Antonio Biondi, Nicolas Desneux, Gaetano Siscaro, Giovanna Tropea Garzia, Edwige Amiens-Desneux, Lucia Zappalà Abstract: Integrated Pest Management (IPM) programs may include pesticide applications, therefore assessing their potential side effects on biocontrol agents is of primary importance for

12 XIV implementing IPM. Bracon nigricans (Hymenoptera: Braconidae) is a larval ectoparasitoid of Lepidoptera species which was recovered on the South American tomato pinworm, Tuta absoluta (Lepidoptera: Gelechiidae), in various Western Palaearctic countries and is a potential biocontrol agent of this pest. We assessed acute toxicity and sublethal effect on fertility of six bioinsecticides, used for controlling T. absoluta, on B. nigricans. The tested chemicals were abamectin, azadirachtin, borax salt plus citrus essential oil, Bacillus thuringiensis var. kurstaki, emamectin benzoate and spinosad. The wasps were exposed for three days to dried pesticide residues on tomato sprouts, at two time intervals after initial treatment. Mortality was daily checked and the number of adult parasitoids emerged was recorded to assess the effects on reproduction. These data were used to calculate reduction coefficients and the pesticides were classified according to the IOBC toxicity categories. The obtained data were then discussed with the results of a previous study, conducted with the same experimental setting, on the generalist predator Orius laevigatus (Heteroptera: Anthocoridae). The insecticides greatly differed in their toxicity as well as in their persistence and the effects varied also between the two biocontrol agents. Our findings would help to optimize future use of the tested insecticides in IPM and organic farming, notably by preventing the possible side effects of the tested pesticides on biological control agents, both naturally present and artificially released. Furtermore, these results stress the need to careful select the chemicals for efficient IPM programs on tomato crops. Influence of insecticide persistence on the survival of the two braconid parasitoids Chelonus inanitus and Aphidius ervi Pilar Medina, Guillermo Manzanares, Raquel Izarra, Angeles Adán, Guy Smagghe, Elisa Viñuela Abstract: An extended laboratory bioassay was conducted to evaluate the toxicity and the duration of the harmful activity of some modern insecticides and the standard deltamethrin to the braconids Aphidius ervi Haliday (aphid parasitoid) and Chelonus inanitus L. (Spodoptera eggs parasitoid). Adults were initially exposed to fresh residues on tomato leaves for three days, and later on, to aged residues in greenhouse. Fresh residues of flonicamid, flubendiamide and spirotetramat were harmless to both parasitoids. Metaflumizone, however, was classified as moderately persistent for C. inanitus (IOBC C) and persistent for A. ervi (IOBC D), due to the reduction on the life span. Deltamethrin was slightly persistent (IOBC, B) and persistent, (IOBC, D) respectively, for the two natural enemies. Advantages of ecological management of leafminers (Diptera: Agromyzidae) in commercial cultivation of crisphead lettuce in southern Brazil Alessandra R. Carvalho, Vanda H. P. Bueno, Diego B. Silva, Joop C. van Lenteren, Lauro L. Petrazzini, Jony E. Yuri Abstract: The objective this work was to evaluate the potential interference of two types of management of leafminer populations and their parasitoids under commercial production conditions of lettuce in plastic tunnels in the state of Minas Gerais, southeast Brazil. The comparison between conventional management using calendar sprays of insecticides, and ecological management with Bacillus thuringiensis, sticky traps and pheromones, showed that ecological management has several advantages over chemical control. In the plastic tunnel with ecological management lower numbers of leafminers emerged, a greater number of parasitoids emerged, the control costs were lower and the commercial value was equivalent to the conventionally managed crop. Field effect of various new insecticides on settled populations of Amblyseius swirskii (Acari: Phytoseiidae) in a cucumber crop grown in a plastic greenhouse in Almería (Spain) I. Colomer, R. M. Heredia, E. Viñuela

13 XV Macrobials for IPM programs Failure of the biological control of Tuta absoluta using the predator Nesidiocoris tenuis in a protected tomato crop: analysis of factors Khaled Abbes, Brahim Chermiti Abstract: Many attempts have been made to biologically control the tomato leafminer Tuta absoluta in Tunisia using the predator Nesidiocoris tenuis in nurseries, greenhouses and open field tomato crops. However, several essays involving the biological control of this pest failed, particularly in greenhouses. The main reasons for these failures were identified by the authors as the over use of insecticides and fungicides, de-leafing after mirid bug introductions and the probable absence of prey when predators were released. A successful method for whitefly and Tuta absoluta control in tomato. Evaluation after two years of application in practice Javier Calvo, Jose Soriano, Karel Bolckmans, Jose E. Belda Abstract: The tobacco whitefly, Bemisia tabaci, and the tomato borer, Tuta absoluta, are major pests of tomato. The mirid bug Nesidiocoris tenuis is an effective natural enemy of whitefly and recently it has been shown to be also a potential biological control agent of T. absoluta. In addition, some parasitoids from the Mediterranean basin have been found attacking T. absoluta and Necremnus artynes is particularly promising. The present study presents the results from experiments conducted to develop a biologically-based management strategy in tomato for whitefly and T. absoluta control and its application in practice thereafter. It first shows the results evaluating an alternative release method for N. tenuis and its combination with N. artynes. It was demonstrated that this alternative release method (pre-plant application) increased control capacity of N. tenuis, provided good control of whitefly and T. absoluta and reduced control costs and the addition of N. artynes did not increase its effectiveness. Although, supplementary releases of this parsite increased the effectiveness of after planting releases of N. tenuis (standar application). Thus, the pre-plant application of N. tenuis alone would be the more efficient method due to it reduced control costs and complexity. Implementation of this strategy has greatly increased the use of biologically-based management strategies in tomato in Spain, and would likely have the same effect in other production areas around the world. IPM strategies in tomato crops in Spanish greenhouses: Effects of cultivars and the integration of natural enemies E. Vila, A. Parra, D. Beltrán, J. R. Gallego, F. J. Fernandez, T. Cabello Abstract: The use of Nesidiocoris tenuis, the main released predator, has contributed to the establishment of biocontrol programmes in more than 50% of the protected tomato crops in the southeast of Spain. Three predator releasing strategies (standard, refuge plants and biopropagation) were evaluated according to the season of the year. The development of the predator on different cultivars was monitored and its combination with Trichogramma achaeae in different seasons discussed. Mechanical release of Phytoseiulus persimilis and Orius laevigatus on protected crops Giovanna Tropea Garzia, Sabina Failla, Giuseppe Manetto, Gaetano Siscaro, Lucia Zappalà Abstract: Mechanical releases of the predatory mite Phytoseiulus persimilis (Acarina: Phytoseiidae) and the predatory bug Orius laevigatus (Heteroptera: Anthocoridae) were carried out in cultivated protected crops (sweet pepper and chrysanthemum) to control the two-spotted spider mite Tetranychus urticae and the western flower thrips, Frankliniella occidentalis. Natural enemies were distributed using a newly developed mechanical device and compared with manual distribution. In all experimental trials carried out with the device the results showed a greater uniformity of distribution, a reduction of release time and ease of application. In most of the mechanised plots the predators were regularly recovered reducing the percentage of leaves infested and their density, so controlling pests sooner than in the manually released ones.