SIDE-EFFECTS OF COMMONLY USED CROP PROTECTION PRODUCTS IN PEAR ON TWO BENEFICIAL MIRIDAE BUGS.

Anthocoris nemoralis, Anthocoris nemorum and Orius spp. are not the only beneficial predatory bugs inhabiting pear orchards in Belgium. Quite often, the Miridae bugs Heterotoma spp. and Pilophorus spp. can be found during spring and summer in these orchards, thereby feeding on several pests such as psyllids, aphids, spider mites, ... . Side-effects are usually assessed on Anthocoris and Orius spp., but due to the potential importance of Miridae bugs in pest reduction, we tested some commonly used crop protection products used in pear cultivation on Heterotoma planicornis and Pilophorus perplexus (residue-based tests in petri-dishes). One day after treatment, mortalities already could be observed for some products. Seven days after treatment, abamectin, deltamethrin and thiacloprid were considered the most toxic products as stated by the IOBC classification. This outcome was then analysed with regard to different treatment schedules, providing insights in potential side-effects on crop protection treatments on the composition of beneficial fauna in pear orchards.

POPULATION DYNAMICS OF DROSOPHILA SUZUKII (DIPTERA: DROSOPHILIDAE) IN BELGIUM IN 2013.

The vinegar fly Drosophila suzukii (Matsumura) (Diptera Drosophilidae), spotted wing drosophila, is a new invasive fruit pest that recently became established in Europe. Unlike other fruit flies that typically only infest overripe and rotten fruit, D. suzukii females oviposit in ripe fruit leading to considerable economic losses of fruit during production. In 2011 and 2012 D. suzukii was detected at several places in Belgium. In 2013, a large-scale monitoring in frame of the project "FLY ALERT" (FOD) was executed using traps with liquid attractant (apple cider vinegar) at more than 100 locations across Belgium during the whole fruit growing season. At 16 locations we also compared the efficacy of a 'bottle type' trap with a 'cup type' trap. The results show that D. suzukii has expanded its distribution in Belgium. Remarkably, in 2013 as well as in 2012 the first detections were made only in the second part of the growing season (August) and the populations reached their peak only at the very end of the season (November). In the bottle type trap the first flies were caught 2-3 weeks earlier than in the cup type trap. In addition, also the population peaks were on average 1 week earlier when monitored with the bottle trap compared to when monitored with the cup trap. In 2014, after an exceptional mild winter adult D. suzukii flies were continuously detected throughout the winter and early spring. The implications of these findings for the phenology of D. suzukii in the Northwest climate region of Europe are discussed.

BIOLOGY AND CONTROL OF THE APPLE MEALYBUG PHENACOCCUS ACERIS (SIGNORET) IN BELGIUM.

Although in general very rare, some outbreaks of the apple mealybug Phenococcus aceris (Signoret) (Hemiptera: Pseudococcidae) were reported in the Belgian fruit growing area recently. This insect is known to be geographically widespread, to have a broad host range and to infest apple trees. Damage at harvest is considerable when sooty molds, a consequence of the pest's honeydew production, cover the fruits. Indirect damage of an infection is caused in cherry cultivation through transmission of the Little cherry virus (LChV2). Efficacy trials were executed in infested apple orchards in the Belgian fruit growing area and the life cycle of the pest on apple was studied more into detail. Apple mealybugs are univoltine, overwinter as 2nd instar nymphs inside a white cocoon on the tree (under the bark, in crevices) and leave their overwintering site in early spring (mid March). On sunny days the nymphs become active, move around and attach to start feeding (mid April). After a final moult into the adult form, females lay eggs in a cocoon-like white structure (from flowering on). Following hatching (end May), massive numbers of young nymphs spread out on the underside of the leaves (mid June) where they feed through suction. In order to manage this pest the efficacy of several plant protection products was tested in two infested apple orchards. Results indicated that mortality was high after an application of compounds belonging to the neonicotinoid insecticides. Different application timings and control strategies are possible, with active nymphs being the most vulnerable life stage. The observed degree of parasitation in our trial orchards also indicates a biological control contribution of parasitic wasps that should be taken into account. A decent IPM-strategy based on our results solved the problem in both apple orchards.

Control of sawflies in apple and pear in Belgium.

In the Belgian fruit growing area, sawflies are generally common but minor pests in pome fruit. Though, intensity and frequency of sawfly damage in apple and pear is increasing the last years in IPM and especially in organic orchards. The main species are apple sawfly (Hoplocampa testudinea Klug) and pear sawfly (Hoplocaompa brevis Klug) and recently also pear shoot sawfly (Janus compressus Fabricius). Here we report efficacy results on all three sawfly species fromtrial of three consecutive years (2011, 2012, and 2013). Flights and embryonic development were monitored and small plot efficacy trials were executed. Control of apple sawfly was complete (97.6% Abbott -trial 2011) when thiacloprid at 120 g/ha LWA was applied at the moment embryos are visible in the sawfly eggs. In 2012, a trial was executed on pear sawfly. Applications with thiacloprid were executed when the embryo was visible in the pear sawfly eggs and earlier at the start of egg laying. At both application timings, 100% Abbott efficacy was reached. A number of other active ingredients were tested at the moment embryos are visible in the sawfly eggs and very interesting efficacy results were reached for thiofanate-methyl, indoxacarb, spinosad, pyrethrins + piperonyl butoxide (PBO) and acetamiprid. In 2013, a preflowering application with pyrethrins + PBO reached the highest control against this pest. The most effective active ingredients of the pear sawfly trial were applied also in a trial on pear shoot sawfly. Efficacies were low or lacking, except for thiacloprid. Thiacloprid is in pear growing in Belgium only registered before flowering and after harvest. Therefore further research is needed to test the effect of earlier applications against this pest. This is a valuable efficacy study on occasionally occurring pests that are able to cause considerable economic losses.

Preliminary results on the ability of pentatomidae to transfer fire blight Erwinia amylovora under controlled conditions.

With their piercing-sucking mouthparts stink bugs (Heteroptera: Pentatomidae), a major pest in especially organic orchards, create wounds in fruit of pear trees. As Erwinia amylovora (Burrill, Winslow et al.), a wide spread bacterial disease affecting many rosaceous plants including pome fruit trees and hawthorn, enters through openings in flowers, leaves, shoots and fruit, feeding punctures caused by these bugs might be inoculated with Erwinia bacteria. In order to investigate the ability of the bugs Pentotoma rufipes L. and Polomena prasina L. to transmit fire blight, insects were caught in an organically managed orchard without fire blight, brought into contact with artificially inoculated immature pear fruit/slices and transferred to healthy, mechanically wounded pear fruit/slices. After an incubation period potential transmission of bacteria was examined by evaluation of symptom expression (necrosis, ooze production). To assess the presence of bacteria on the exoskeleton of the tested bugs, all bugs were forced to walk on a semiselective nutrient agar medium. In another experiment the viability of Ea on the exoskeleton was tested -after previous contact with ooze- through washing and plating of the wash water. All experiments were conducted under optimal climatological conditions and according to quarantine standards. Results demonstrated the ability of stink bugs to transfer E. amylovora to fruit and the viability of bacteria on stink bugs externally - both under lab conditions.

Biological efficacy evaluation of mating disruption against the grape berry moth, Lobesia botrana, in grape in glasshouses.

The grape berry moth, Lobesia botrana, is an important pest of grapes. In Belgium, severe losses of production appeared due to the presence of L. botrona in grapes commercially grown in glasshouses. In 2010 mating disruption became a key pest management tactic in Flemish pip fruit orchards after being tested for several years. In other (European) countries, mating disruption is already applied in grapes but there is no registration in Belgium for use in grapes. Searching for an interesting residue free technique for application in small glasshouses, we evaluated the effect of mating disruption for control of L. botrana under protected conditions (glasshouse trials) during three consecutive years (2010-2012). A prerequisite for successful mating disruption is the treatment of large contiguous areas, as the pheromone product will not remain in sufficient quantities on narrow sites and, moreover, a larger portion of the crop remains at risk from migrating mated females. The glasshouses are rather small, however, we hypothesized that these so called border effects are not present due to the secluded spaces of the glasshouses. Pheromone dispensers were deployed and flights of L botrano moths were monitored (pheromone traps/UV-traps) during the whole time period of the trials. In the mating disrupted glasshouses hardly any moth was caught anymore in pheromone traps, in contrast to UV-traps or catches by pheromone traps in comparable non-mating disrupted glasshouse compartments. This indicates the incapability of the males to localize sex pheromone sources, and hence, the inability of finding females for mating. In line with these flight monitoring data, the nearly absence or strongly decreased L. botrana caused damage symptoms in the mating disrupted glasshouses demonstrates the benefits of the mating disruption technique under protected conditions of grape growing in glasshouses.

Consistent codling moth population decline by two years of mating disruption in apple: a Flemish case study.

Codling moth (Cydia pomonella) is one of the most important pests in apple and pear. In 2010 mating disruption became a key pest management tactic in Flemish pip fruit orchards, largely due to a government subsidy and demonstrating projects aiming to widen the area treated by pheromones as large as possible. As a consequence, the mating disruption strategy was applied at approximately 7.500 ha, or half of the pip fruit area, in 2010 and 2011. The sudden large-scale implementation of this technique changed the codling moth management landscape. Here we present a case study of a commercially managed orchard that suffered from high codling moth pressures for many years, as did the surrounding area. The RAK3 mating disruption system was introduced at this location in 2010, and was continued in 2011. Systematic detailed codling moth flight data for this location are available for many years. In addition, comprehensive data on damage levels of chemically untreated windows spread all over the test orchard in a randomized block design were obtained in successive years, enabling us to thoroughly evaluate the effect of the changed codling moth management strategy. Data from 2011 included damage levels in chemically treated windows when the entire orchard was applied once at the flight peak of Cydia pomonella. In 2009, before introduction of mating disruption, a mean of 8.25 +/- 5.54% of the fruits were infested at harvest when assessed in completely untreated windows. After two years of mating disruption, supported with a full chemical support in 2010, except for the untreated assessment windows, and only one application on the flight peak of 2011, damage was reduced to less than 0.03% at harvest. This is a valuable case study to demonstrate the benefits of the mating disruption approach.

Earwig management tool: an IPM decision aid system for augmentation of European earwig populations (Dermaptera: Forficulidae) in pip fruit orchards.

A key element of integrated pest management (IPM) is the suppression of potential pest outbreaks by beneficial arthropods. The European earwig, Forficula auricularia L., is an important natural enemy of a wide range of insect pests in pip fruit orchards. However, earwig population sizes vary greatly from location to location, illustrating their sensitivity to biotic and abiotic factors, especially human interventions relating to orchard management. In order to help growers sparing and augmenting earwig populations in their pip fruit orchards, we developed a software tool that integrates a sophisticated earwig phenology model with management recommendations. The program is based on a day degree model for earwigs which is fed by temperature data collected by the pcfruit research centre. In addition, a pesticide database with known side effects of a wide range of products on the different life stages of earwigs is integrated in the system. The output gives the current status of the earwig population and management recommendations for activities critical for their survival. Hence, by consultation of this user-friendly software fruit growers can predict the earwig development in the field at any time, and organize the timing of orchard management actions taking into account the presence of (sensitive) life stages of the earwig life cycle. Doing so, negative effects specific orchard management actions, such as badly timed spray applications and soil tillage, can be avoided.

Life cycle and control of the forest bug Pentatoma rufipes L. in organically managed pear orchards.

In Belgian pear production growers may encounter major economic problems when fruit becomes damaged by bugs. One of the major pests belongs to the stink bugs, Pentatoma rufipes L. (Heteroptera, Pentatomidae) which feeds by piercing into buds, flowers and fruit. To study the population dynamics of this species it was monitored in organic pear orchards by the beating tray method. Results revealed their univoltine life cycle and their presence within the orchard during the entire year. They overwinter in the second nymphal stage and become active in March/April. After 3 moultings adulthood is reached during May/June. In summer mating takes place and eggs are laid at the underside of the leaves of the fruit trees. By the end of August the eggs hatch and the first young nymphs appear. At first they aggregate but when they have moulted into the next stage dispersal takes place (September) and by October they search for hiding places on the trees to hibernate. As damage is done before and after blossom the application of an insecticide has to be well defined and timed. Therefore the efficacy of spinosad and natural pyrethrum, applied once or twice before and/or after bloom, was compared in a field trial. Data indicated that a single application of spinosad after bloom and a double application of spinosad or pyrethrum (before and after bloom) reduced the number of bugs with 83.3%. Damage to the crop was the lowest (4.5%) when spinosad was applied twice but no statistical difference with both other applications was found.

Optimal profit of the parasitation by Aphelinus mali in an IPM complementary strategy for the control of Eriosoma lanigerum.

During summer the parasitoid Aphelinus mali may certainly reduce the infestation of woolly apple aphid (Eriosoma lanigerum), but studies on the single interaction rarely indicate sufficient biological control in the period May-June. In this period chemical control by spirotetramat or pirimicarb remains indispensable in order to anticipate on dense migration waves and subsequent colonization of extension shoots by E. lanigerum. The limited parasitation by A. mali around flowering is linked with a delayed emergence from diapause and with a slower reproduction rate than its host. In 2010 and 2011 the first adult flights monitored on yellow sticky traps corresponded perfectly with the currently used prediction models for A. mali. Further accurate monitoring all along the season enabled also to determine a well defined endo-parasitic phase of A. mali occurring after the small peak observed around flowering. During this endo-parasitic phase A. mali larvae reside inside their mummified host. Compounds with higher acute toxicity on A. mali adults, like chloronicotinyl insecticides (CNI's), are preferably positioned here. Selectivity in the time can then be claimed. Respecting this principle, the further parasitation potential of A. mali in summer is not hampered. Preservation of the first peak of flights of A. mali in the pre-flowering period is essential for an exponential flight increase. This is essential for the parasitation of E. lanigerum in summer, which constitutes a valuable complement in the integrated control strategy.

Integrated management of phytoplasma diseases in pome fruit: an overview of efficacy results of IPM insecticides against pear Psylla (Cacopsylla pyri).

Phytoplasmas are plant pathogenic mollicutes that cause devastating diseases in various crops worldwide. The closely related pome fruit tree phytoplasmas Candidatus Phytoplasma mali and Candidatus Phytoplasma pyri are the causal agents of apple proliferation and pear decline, respectively. They can be transmitted from tree to tree by Psyllidae. As pear suckers (Cacopsylla pyri) are widely considered to be the most important pest in pear orchards, a good control of this insect vector is a key element for limiting the natural spread of pear decline. Efficient control relies on a perfect tuning of treatment schedules, taking into account efficacies of (at preferably) low-impact insecticides and side-(repellent)-effects of alternative products (e.g. kaolin, mineral oils and fungicides), the optimal positioning of these crop protection agents, and the best possible presence of beneficial predators. The department of Zoology of the pcfruit vzw research institute (Belgium) has a long tradition of executing insecticide field trials according to EPPO guidelines. Here, we present an overview of the results of a selection of IPM-compatible insecticides (abamectin, spirotetramat, thiacloprid, spinosad, spirodiclofen), tested in efficacy trials against pear Psylla on different life stages during the last decade. Based on these results and monitoring data of pest and beneficial biology, we additionally propose optimal pear Psylla control schedules which allow to reduce the number of (phytoplasma harbouring) psyllids in integrated systems to a minimum.

Effects of sublethal doses of crop protection agents on honey bee (Apis mellifera) global colony vitality and its potential link with aberrant foraging activity.

Honey bees (Apis mellifera) are the most economically valuable pollinators of fruit crops worldwide. Taking into account bees' contributions to other flowering agricultural crops, about one-third of our total diet comes directly or indirectly from bee-pollinated plants. However, in recent years there increasingly have been worrisome alarm sounds on serious bee mortalities and mysterious disappearance of bees from beehives. Among several environmental factors (e.g. climate and bee pathogens), stress factors arising from agricultural practices can potentially play a role in bee losses. Detailed knowledge on the effects of plant protection products is essential to improve usage with minimal risks. In order to identify potential medium- and long-term effects, we followed up various sublethal contaminated hives during the prolongation of the fruit-growing season. More specifically, a large-scale experiment was conducted in which at four distinct locations (in the Limburg region of Belgium) four different bee colonies (representing three different contaminations -imidacloprid, fenoxycarb, indoxacarb- and a non-contaminated control hive) were thoroughly monitored every 2-7 days. Our observations point towards decays of overall colony vitality for several hives a couple of weeks after treatment, as indicated by a set of carefully assessed parameters including the total amount of active and dead bees, total surface of capped brood and overall colony weight. These outcomes could be linked to subtle differences in foraging activity between distinct hives. The implications of these results are discussed in terms of potential short-term and long-term consequences of disturbed foraging ability triggered by exaggerated exposure to sublethal doses of crop protection chemicals, and its potential impact on colony health.

The selectivity in the time of thiacloprid (Calypso 480 SC) applied at preblossom on the further Anthocoris nemoralis population build up in pears.

Predatory bugs (Anthocoris sp.) are key predators of the pear sucker (Psyllo pyri) in pear orchards. In order to enhance biological control it is important to verify if crop protection products applied against Psylla pyri display potential short- or long-term detrimental effects on Anthocoris populations. This study focuses on the impact of thiacloprid (Calypso 480 SC), applied at preblossom for the control of the first generation pear sucker Psylla pyri, on subsequent population build up of Anthocoris nemoralis later in the season. Hereto, five large-scale trials on pear Conference were conducted. Two types of orchards were selected: the first type is surrounded by abundant vegetation from which Anthocorids easily can fly in when attracted by the presence of prey (Psylla pyri); the second type has no "reservoir" vegetation around the orchard and here it is considered that mostly the residential population of Anthocorids constitutes the predation. In all five trials positioning of thiacloprid preblossom did not prevent the normal population build up of the predatory Anthocoris sp. population later in the season. As both in orchards with surrounding vegetation and in orchards without adjacent vegetation a regular build up of the Anthocoris sp. population was observed, the outcomes of this study indicate that a preblossom application of Calypso 480 SC can be considered as "selective in time" for further Anthocoridae dynamics.

Side effects of plant protection products and biological interactions on the European earwig Forficula auricularia L.

Plant protection products are designed to control pests but can have negative side effects on non-target arthropods thus disturbing the important population of natural enemies required for biological control. Although the European earwig, Forficula auricularia L, (Dermaptera: Forficulidae) is not considered to be a key beneficial in pome fruit, it is an important predator of several pests, e.g. woolly apple aphid and pear sucker. The impact of non-selective plant protection products at crucial moments in their (univoltine) life cycle can be of significant relevance compared to insects with numerous generations. Foliar applications in spring when nymphs are migrating into the trees can reduce the number of adults in summer and subsequently affect the population size next year. Multiple and/or combined spraying during summer on adults may have a cumulative effect resulting in less over wintering females which possibly exhibit poor reproductive performance. Previous residual contact bioassays already revealed the harmful side effect of several formulated products on adults. Insects showing sub lethal symptoms recovered partially or died eventually. As spinosad caused significant toxicological effects it was subsequently tested in 3 different dose rates on adults and N4-nymphs. We noticed not only a clear dose-response relationship but N4-nymphs proved to be more susceptible than adults; even a dilution of 1/9-th of the registered dose rate still caused a mortality of 45.5 % after 20 days. Understanding the earwig's population dynamics is essential for efficient practical biocontrol. It proves difficult to increase population levels to sufficient high numbers for optimal pest control. Local biological factors might be limiting. Therefore, we tested two hypotheses that pertain to population limitation: 1. Bird predation during summer, 2. Small mammal nest predation during winter. Enclosure experiments showed no negative bird effect on earwig densities unless large bird flocks inhabited the area. Small mammals did not actively predate the over wintering nests, although other predatory arthropods may be important.

Functional display of family 11 endoxylanases on the surface of phage M13.

Two family 11 endoxylanases (EC 3.2.1.8) were functionally displayed on the surface of bacteriophage M13. The genes encoding endo-1,4-xylanase I from Aspergillus niger (ExlA) and endo-1,4-xylanase A from Bacillus subtilis (XynA) were fused to the gene encoding the minor coat protein g3p in phagemid vector pHOS31. Phage rescue resulted in functional monovalent display of the enzymes as was demonstrated by three independent tests. Firstly, purified recombinant phage particles showed a clear hydrolytic activity in an activity assay based on insoluble, chromagenic arabinoxylan substrate. Secondly, specific binding of endoxylanase displaying phages to immobilized endoxylanase inhibitors was demonstrated by interaction ELISA. Finally, two rounds of selection and amplification in a biopanning procedure against immobilized endoxylanase inhibitor were performed. Phages displaying endoxylanases were strongly enriched from background phages displaying unrelated proteins. These results open perspectives to use phage display for analysing protein-protein interactions at the interface between endoxylanases and their inhibitors. In addition, this technology should enable engineering of endoxylanases into novel variants with altered binding properties towards endoxylanase inhibitors.