Potential Management Options for the Invasive Moth Spodoptera frugiperda in Europe.

We here review and discuss management options that growers in Europe could take in response to the expected invasion of the fall armyworm, Spodoptera frugiperda (Lepidoptera: Noctuidae). The focus is put on maize but the information provided is also relevant for other crops potentially affected. A sound forecasting system for fall armyworm both on a regional as well as at local scale should be established to alert growers as early as possible. Whilst a number of cultural control methods are adopted by maize growers in different regions globally to fight fall armyworm, many of them may either not be highly effective, too laborious, or otherwise unfeasible within the mechanized crop production systems used in Europe. Potential is seen in the stimulation of natural enemies through conservation biocontrol approaches, e.g., the planting of flower strips or intermediate cover crops, reducing tillage intensity, and avoiding broad-spectrum insecticides. To manage fall armyworm infestations, several effective biologically-based products are available globally, and some in Europe, e.g., based on specific baculoviruses, certain Bacillus thuringiensis strains, few entomopathogenic nematodes, and a number of botanicals. These should be given priority to avoid a major influx of insecticides into the maize agro-ecosystem once the fall armyworm arrives and in case growers are not prepared. Plant protection companies, particularly biocontrol companies should act proactively in starting registration of ingredients and products against fall armyworm in Europe. European maize growers should be made aware, in time, of key features of this new invasive pest and appropriate control options.

Selection of the Most Effective Trichogramma Strains (Hymenoptera: Trichogrammatidae) From Myanmar to Control Asian Corn Borer, Ostrinia furnacalis (Lepidoptera: Crambidae).

Selecting the candidate agents or species is a fundamental step in developing effective biological control programs. However, no attempts have been made to evaluate the efficacy of Trichogramma Westwood (Hymenoptera: Trichogrammatidae) strains against the Asian corn borer, Ostrinia furnacalis Guenée (Lepidoptera: Crambidae) in Myanmar. In this study, Trichogramma strains were investigated through a series of glass tube bioassays and under a semifield condition. Ostrinia furnacalis or Corcyra cephalonica egg preferences were compared among six strains of Trichogramma ostriniae, three of Trichogramma chilonis, and four of Trichogramma dendrolimi using a choice-test assay design. Significant differences were observed at the inter- and intraspecific levels. Four strains of T. ostriniae and two of T. dendrolimi showed a strong preference for O. furnacalis eggs, while two strains of T. chilonis and one T. dendrolimi strain preferred Corcyra cephalonica Stainton (Lepidoptera: Pyralidae) eggs. The remaining strains showed no preference. Eleven strains were examined on O. furnacalis eggs only, wherein parasitism, sex ratio, and total progeny per female were highest for two strains of T. ostriniae (respectively: 67.6 ± 3.0%, 82.7 ± 2.3%, 49.6 ± 2.8, and 67.6 ± 3.6%, 90.0 ± 2.4%, 42.7 ± 2.6), and one strain of T. dendrolimi (65.2 ± 3.2%, 84.7 ± 2.6%, 46.3 ± 2.8). Parasitism and searching capacity were evaluated for the three, best performing strains in cages containing maize plants. One T. ostriniae from Southern Shan State, Myanmar parasitized ~81% of egg masses, and should be considered a candidate biological control agent against O. furnacalis in Myanmar.

Assessing the Potential of Inoculative Field Releases of Telenomus remus to Control Spodoptera frugiperda in Ghana.

In response to the threat caused by the fall armyworm to African maize farmers, we conducted a series of field release studies with the egg parasitoid Telenomus remus in Ghana. Three releases of ≈15,000 individuals each were conducted in maize plots of 0.5 ha each in the major and minor rainy seasons of 2020, and compared to no-release control plots as well as to farmer-managed plots with chemical pest control. No egg mass parasitism was observed directly before the first field release. Egg mass parasitism reached 33% in the T. remus release plot in the major rainy season, while 72-100% of egg masses were parasitized in the minor rainy season, during which pest densities were much lower. However, no significant difference in egg mass parasitism was found among the T. remus release plots, the no-release control plots and the farmer-managed plots. Similarly, no significant decrease in larval numbers or plant damage was found in the T. remus release fields compared to the no-release plots, while lower leaf and tassel damage was observed in farmer-managed plots. Larval parasitism due to other parasitoids reached 18-42% in the major rainy season but was significantly lower in the minor rainy season, with no significant differences among treatments. We did not observe significant differences in cob damage or yield among the three treatments. However, the lack of any significant differences between the release and no-release plots, which may be attributed to parasitoid dispersal during the five weeks of observation, would require further studies to confirm. Interestingly, a single application of Emamectin benzoate did not significantly affect the parasitism rates of T. remus and, thus, merits further investigation in the context of developing IPM strategies against FAW.

Molecular and Morphological Identification of Trichogramma (Hymenoptera: Trichogrammatidae) Species From Asian Corn Borer (Lepidoptera: Crambidae) in Myanmar.

The Asian corn borer, Ostrinia furnacalis (Guenée), is the most devastating maize pest in Myanmar. The damage caused by this pest has become a significant obstacle to farmers' income and the national economy. Control of O. furnacalis is challenging due to its protected feeding sites and the crop's height during later phases of the pest's attack. Trichogramma (Hymenoptera: Trichogrammatidae) parasitoids have been used successfully against corn borers in other countries, but knowledge on the species composition in Myanmar is limited. As a first step for potential biological control of Asian corn borer in Myanmar, Trichogramma were collected from major maize growing regions. Identification was performed based on both morphological and molecular techniques. The majority of collected specimens were identified as Trichogramma ostriniae Pang and Chen, T. chilonis Ishii, and T. dendrolimi Matsumura. This is the first report on the composition of Trichogramma species, and an important step towards the establishment of biological control against O. furnacalis in Myanmar.

Next-generation biological control: the need for integrating genetics and genomics.

Biological control is widely successful at controlling pests, but effective biocontrol agents are now more difficult to import from countries of origin due to more restrictive international trade laws (the Nagoya Protocol). Coupled with increasing demand, the efficacy of existing and new biocontrol agents needs to be improved with genetic and genomic approaches. Although they have been underutilised in the past, application of genetic and genomic techniques is becoming more feasible from both technological and economic perspectives. We review current methods and provide a framework for using them. First, it is necessary to identify which biocontrol trait to select and in what direction. Next, the genes or markers linked to these traits need be determined, including how to implement this information into a selective breeding program. Choosing a trait can be assisted by modelling to account for the proper agro-ecological context, and by knowing which traits have sufficiently high heritability values. We provide guidelines for designing genomic strategies in biocontrol programs, which depend on the organism, budget, and desired objective. Genomic approaches start with genome sequencing and assembly. We provide a guide for deciding the most successful sequencing strategy for biocontrol agents. Gene discovery involves quantitative trait loci analyses, transcriptomic and proteomic studies, and gene editing. Improving biocontrol practices includes marker-assisted selection, genomic selection and microbiome manipulation of biocontrol agents, and monitoring for genetic variation during rearing and post-release. We conclude by identifying the most promising applications of genetic and genomic methods to improve biological control efficacy.

The Efficacy of Alternative, Environmentally Friendly Plant Protection Measures for Control of Fall Armyworm, Spodoptera Frugiperda, in Maize.

The invasive fall armyworm (FAW) is threatening maize production and the livelihoods of millions of smallholder farmers in the newly invaded areas in Africa and Asia. To control this new key pest and to overcome health, environmental, and resistance problems related to the indiscriminate use of insecticides, effective and sustainable alternative pest control approaches are needed. Here, we report on field trials that tested maltodextrin, neem-based products, ash, and soil, as well as the locally produced alata samina soap, in the Upper West and Greater Accra regions, Ghana. Significant reductions of larval numbers and crop damage, together with increased yields, were mostly achieved by applying the insecticide emamectin benzoate, which was considered the positive control in this set of trials. However, high efficiency and cost-benefit ratios were also achieved with two neem-based products. Maltodextrin was only efficient at one of the two sites, with a clear dose-dependent effect, while the higher dosage was nearly as effective as emamectin benzoate. Due to its relatively high product cost, maltodextrin is generally less cost-efficient. Ash and soil, as well as alata samina soap treatments, did not efficiently reduce FAW larval numbers or crop damage at the dosages tested; thus, they also did not significantly increase maize yields.

Impact Assessment of Biological Control-Based Integrated Pest Management in Rice and Maize in the Greater Mekong Subregion.

The impact and sustainability of two interventions that have been formulated to introduce integrated pest management (IPM) into rice and maize crops in Southwestern China, Laos, and Myanmar between 2011 and 2016, and were assessed at the end of 2017. From 22 Trichogramma rearing facilities established during the interventions, 11 were still producing substantial quantities of biocontrol agents 1.5 years after project support had ended, while seven had stopped operations completely, and four were doing stock rearing only. Through the implementation of biological control-based IPM, slightly higher yields were achieved in maize and rice (4-10%), when compared to control farmers, but the difference was not statistically significant. However, the use of pesticides nearly halved when farmers started using Trichogramma egg-cards as a biological control agent. Support from either public or private institutions was found to be important for ensuring the sustainability of Trichogramma rearing facilities. Many of the suggested IPM measures were not adopted by smallholder farmers, indicating that the positive impacts of the interventions mostly resulted from the application of Trichogramma biological control agents. The following assessment suggests that further promotion of IPM adoption among farmers is needed to upscale the already positive effects of interventions that facilitate reductions in synthetic pesticide use, and the effects on sustainable agricultural production of rice and maize in the target area more generally.

Can Anastatus bifasciatus Be Used for Augmentative Biological Control of the Brown Marmorated Stink Bug in Fruit Orchards?

The generalist egg parasitoid Anastatus bifasciatus (Geoffroy) (Hymenoptera: Eupelmidae) is the most prevalent egg parasitoid of the invasive Halyomorpha halys (Stål) (Hemiptera: Pentatomidae) in Europe. To assess its efficacy against the pest H. halys and to validate the potential risks for non-target species in a realistic field setting, inundative releases were conducted over three consecutive years in four fruit orchards in Switzerland and Italy. In total, more than 4300 A. bifasciatus females were released, which was equivalent to 11,000 to 26,000 females per hectare, depending on distances between trees in each orchard. Parasitism of freeze-killed sentinel H. halys eggs achieved with the current release strategy was on average 6% (range: 2%-16%) and considered not high enough to effectively suppress the pest. However, the overall impact of A. bifasciatus on the mortality of H. halys eggs was likely underestimated. If pre-imaginal parasitoid mortality (3.3%) and host feeding (6%) are added to the observed parasitism (6%), the actual induced mortality of H. halys eggs may reach more than 15%. Parasitism of lepidopteran non-target species reached an average of 8% and thus, some degree of non-target parasitism after mass releases may be expected. To quantify the impact of the parasitoids in the orchards more precisely, naturally laid egg masses should be used in future trials to include host-finding cues of the host and host plants, and larger scale releases with potentially higher densities of parasitoids should be considered.

Using the egg parasitoid Anastatus bifasciatus against the invasive brown marmorated stink bug in Europe: can non-target effects be ruled out?

The brown marmorated stink bug, Halyomorpha halys (Stål) (Hemiptera: Pentatomidae), has been causing massive damage to various fruit and vegetable crops after its arrival in the USA, and more recently in Europe. To provide an alternative control measure to pesticides, the native egg parasitoid Anastatus bifasciatus (Geoffroy) (Hymenoptera: Eupelmidae) was considered as a candidate biological control agent for inundative releases in Europe. In the risk assessment study presented here, all nine heteropteran and 14 out of 19 tested lepidopteran non-target species produced viable A. bifasciatus offspring. The proportion of A. bifasciatus females producing offspring did not differ between non-target and target for 19 out of the 28 non-target species. Larger host eggs corresponded to increased female-biased sex ratio of the offspring as well as an increase in size, particularly for females, with hind tibia lengths varying from 645.5 ± 46 to 1084 ± 28.5 µm. Larger females were also found to have higher offspring production and increased life expectancy. The results of this study confirmed the polyphagous nature of A. bifasciatus and suggest that a number of non-target species, including Lepidoptera of conservation interest, may be attacked in the field. Thus, non-target effects cannot entirely be ruled out, but more information is needed from semi-field and field studies to fully assess potential environmental risks due to inundative releases of this native parasitoid.

Assessment of Trichogramma japonicum and T. chilonis as Potential Biological Control Agents of Yellow Stem Borer in Rice.

Two species of Trichogramma wasps were assessed for their effectiveness against yellow stem borer Scirpophaga incertulas. A laboratory cage test with T. japonicum and T. chilonis showed that both species parasitized yellow stem borer egg masses at 60.0% ± 9.13% and 40.7% ± 7.11%, respectively, with egg parasitism rates of 15.8% ± 22.2% for T. japonicum and 2.8% ± 5.0% for T. chilonis. Once the host eggs were parasitized, emergence rates were high for both species (95.7% ± 0.12% for T. japonicum and 100% for T. chilonis). In paddy field trials, the two Trichogramma species were released at three densities (50,000/ha, 100,000/ha and 200,000/ha) in Southwestern China. Egg mass parasitism was 9% ± 7.7% for T. japonicum and 15% ± 14.1% for T. chilonis, and again only a relatively small fraction of eggs was successfully parasitized. No clear conclusion could be drawn on the most efficient release rate as no significant differences were found among the three release rates. A comparison of field-collected T. japonicum with T. japonicum and T. chilonis mass reared on Corcyra cephalonica showed significantly larger body size and ovipositor length in field-collected wasps, suggesting potentially higher effectiveness on yellow stem borer eggs after at least one generation on the target host. Factors contributing to the low field parasitism rates are discussed.

Toxicity of Insecticides Targeting Rice Planthoppers to Adult and Immature Stages of Trichogramma chilonis (Hymenoptera: Trichogrammatidae).

Planthopper-targeting insecticides, pymetrozine, thiamethoxam, buprofezin, and nitenpyram, were tested under laboratory conditions for toxicity to adults and immatures of Trichogramma chilonis Ishii, using standard tests described by International Organization for Biological Control (IOBC). In the dry film residue test, all insecticides resulted in >90% mortality in T. chilonis adults and were ranked as moderately harmful. Persistent toxicity tests revealed that nitenpyram was short-lived and the other three insecticides were of slightly persistent toxicity to the wasp adults. Effects of the insecticides on egg, larval, and prepupal stages of T. chilonis were investigated with striped stem borer as host. At the three stages of T. chilonis (within the host egg), all the insecticides reduced parasitism rate, but nitenpyram and pymetrozine applied at egg stage, buprofezin and nitenpyram at larval stage, and buprofezin and thiamethoxam at prepupal stage of T. chilonis reduced parasitism by <30% in comparison with the control, and were thus ranked as harmless. Although insecticide treatment of the three immature stages of T. chilonis all reduced wasp emergence from host eggs, only thiamethoxam applied at larval stage and buprofezin at prepupal stage resulted in >30% reduction in emergence rate as compared with the control and were categorized as harmful. Immature duration of T. chilonis was only significantly extended by nitenpyram applied to egg stage than the control. Sex ratio of emerged wasps was not affected by the treatment to immature stages. The data are of significance for IPM programs incorporating inundative release of T. chilonis for control of lepidopteran rice pests where there is heavy co-occurrence of planthoppers.

Evaluation for potential Trichogramma (Hymenoptera: Trichogrammatidae) strains for control of the striped stem borer (Lepidoptera: Crambidae) in the Greater Mekong Subregion.

Trichogramma species and strains differ significantly in host specificity and performance. Nine Trichogramma strains, six of them collected from paddy fields in the Greater Mekong Subregion, were evaluated for performance on eggs of the striped stem borer, Chilo suppressalis (Walker), in both laboratory and field tests to determine potential Trichogramma strains that can be used in an inundative release in an integrated pest management program. In the laboratory glass vial tests, all strains showed higher parasitism rates on 0-24-h eggs than on the two older age groups (24-48 and 48-72 h). Wasp emergence rate was also higher from parasitized 0-24-h striped stem borer eggs, while Trichogramma immature duration was significantly prolonged on 0-24-h striped stem borer eggs. Parasitism rates differed among Trichogramma strains, with Trichogramma chilonis Ishii CJ strain showing significantly higher parasitism rate than any other strains. In the field tests, parasitism of sentinel striped stem borer eggs by Trichogramma strains released at 50,000, 100,000, and 200,000 wasps per hectare was low, with marginal yet significant differences between strains. The highest parasitism was achieved by T. chilonis CJ strain at the high and medium release rates. Hence, it can be concluded that T. chilonis CJ strain released at 100,000 wasps per hectare may be a cost-effective control tactic for field releases targeting striped stem borer.

Honeydew feeding in the solitary bee Osmia bicornis as affected by aphid species and nectar availability.

Like honey bees (Apis mellifera), non-Apis bees could exploit honeydew as a carbohydrate source. In addition to providing carbohydrates, this may expose them to potentially harmful plant products secreted in honeydew. However, knowledge on honeydew feeding by solitary bees is very scarce. Here we determine whether the polylectic solitary bee Osmia bicornis (=O. rufa) collects honeydew under semi-field conditions, and whether this is affected by aphid species and presence of floral nectar. Bees were provided with oilseed rape plants containing flowers and/or colonies of either Myzus persicae or Brevicoryne brassicae. We used the total sugar level of the bee crop as a measure of the individual's nutritional state and the oligosaccharide erlose as indicator for honeydew consumption. Erlose was present in honeydews from both aphid species, while absent in oilseed rape nectar, nor being synthesized by O. bicornis. When bees were confined to a single honeydew type as the only carbohydrate source, consumption of M. persicae honeydew was confirmed for 47% of the bees and consumption of B. brassicae honeydew for only 3%. Increased mortality in the latter treatment provided further evidence that B. brassicae honeydew is an unsuitable food source for O. bicornis. All bees that were given the choice between honeydew and floral nectar showed significantly increased total sugar levels. However, the fact that no erlose was detected in these bees indicates that honeydew was not consumed when suitable floral nectar was available. This study demonstrates that honeydew exploitation by O. bicornis is dependent on honeydew type and the presence of floral nectar.

Impact of transgenic oilseed rape expressing oryzacystatin-1 (OC-1) and of insecticidal proteins on longevity and digestive enzymes of the solitary bee Osmia bicornis.

The risk that insect-resistant transgenic plants may pose for solitary bees was assessed by determining longevity of adult Osmia bicornis (O. rufa) chronically exposed to transgenic oilseed rape expressing oryzacystatin-1 (OC-1) or to the purified insecticidal proteins recombinant rOC-1, Kunitz soybean trypsin inhibitor (SBTI), Galanthus nivalis agglutinin (GNA), or Bacillus thuringiensis toxin Cry1Ab dissolved in sugar solution (at 0.01 and 0.1%, w:v, Cry1Ab only at 0.01%). Compared to control bees, longevity was significantly reduced by SBTI and GNA at both concentrations and by rOC-1 at 0.1%, but not by Cry1Ab or rOC-1 at 0.01%. Longevity on the OC-1 oilseed rape was not significantly different from the control plants. The effects of SBTI and rOC-1 on longevity were investigated through characterization of the digestive proteinases of O. bicornis and analysis of the response in proteinase profiles to ingestion of these proteinase inhibitors. A relatively complex profile of at least four types of soluble proteolytic enzymes was identified. Serine proteinases were found to be predominant, with metallo and especially cysteine proteinases making a smaller albeit significant contribution. The compensatory response to in vivo enzyme inhibition was similar for SBTI and rOC-1 although less pronounced for rOC-1. It consisted of a non-specific overproduction of native proteinases, both sensitive and insensitive, and the induction of a novel aspartic proteinase.

Potential effects of oilseed rape expressing oryzacystatin-1 (OC-1) and of purified insecticidal proteins on larvae of the solitary bee Osmia bicornis.

Despite their importance as pollinators in crops and wild plants, solitary bees have not previously been included in non-target testing of insect-resistant transgenic crop plants. Larvae of many solitary bees feed almost exclusively on pollen and thus could be highly exposed to transgene products expressed in the pollen. The potential effects of pollen from oilseed rape expressing the cysteine protease inhibitor oryzacystatin-1 (OC-1) were investigated on larvae of the solitary bee Osmia bicornis (= O. rufa). Furthermore, recombinant OC-1 (rOC-1), the Bt toxin Cry1Ab and the snowdrop lectin Galanthus nivalis agglutinin (GNA) were evaluated for effects on the life history parameters of this important pollinator. Pollen provisions from transgenic OC-1 oilseed rape did not affect overall development. Similarly, high doses of rOC-1 and Cry1Ab as well as a low dose of GNA failed to cause any significant effects. However, a high dose of GNA (0.1%) in the larval diet resulted in significantly increased development time and reduced efficiency in conversion of pollen food into larval body weight. Our results suggest that OC-1 and Cry1Ab expressing transgenic crops would pose a negligible risk for O. bicornis larvae, whereas GNA expressing plants could cause detrimental effects, but only if bees were exposed to high levels of the protein. The described bioassay with bee brood is not only suitable for early tier non-target tests of transgenic plants, but also has broader applicability to other crop protection products.

Bacterial community structures in honeybee intestines and their response to two insecticidal proteins.

In this study, the effects of the Bt-toxin Cry1Ab and a soybean trypsin inhibitor (SBTI) on intestinal bacterial communities of adult honeybees (Apis mellifera) were investigated. It was hypothesized that changes in intestinal bacterial communities of honeybees may represent a sensitive indicator for altered intestinal physiology. Honeybees were fed in a laboratory set-up with maize pollen from the Bt-transgenic cultivar MON810 or from the non-transgenic near isoline. Purified Cry1Ab (0.0014% w/v) and SBTI (0.1% or 1% w/v) represented supplementary treatments. For comparison, free-flying honeybees from two locations in Switzerland were analysed. PCR-amplification of bacterial 16S rRNA gene fragments and terminal restriction fragment length polymorphism analyses revealed a total of 17 distinct terminal restriction fragments (T-RFs), which were highly consistent between laboratory-reared and free-flying honeybees. The T-RFs were affiliated to Alpha-, Beta-, and Gammaproteobacteria, to Firmicutes, and to Bacteriodetes. Neither Bt-maize pollen nor high concentrations of Cry1Ab significantly affected bacterial communities in honeybee intestines. Only the high concentration of SBTI significantly reduced the number of T-RFs detected in honeybee midguts, a concentration that also increases bee mortality. Therefore, total bacterial community structures may not be a sensitive indicator for providing evidence for the impact of insecticidal proteins on honeybees at sublethal levels.

Consumption of snowdrop lectin (Galanthus nivalisagglutinin) causes direct effects on adult parasitic wasps.

Honeydew is a common sugar-rich excretion of aphids and other phloem-feeding insects and represents the primary sugar in many agricultural systems. When honeydew-producing insects feed on genetically modified plants, the honeydew can contain amounts of the transgene product. Here we address whether this route of exposure poses a risk for non-target insects. Three species of parasitic wasps were selected: i.e. Aphidius colemani, Trichogramma brassicae and Cotesia glomerata, all of which are known to use honeydew as a carbohydrate source in the field. Wasps were fed sucrose solutions with varying concentrations of snowdrop lectin ( Galanthus nivalisagglutinin, GNA), a protein that has been engineered into crops to confer resistance against homopteran pests and that has been detected in honeydew. Parameters evaluated included gustatory response, longevity, fecundity, progeny emergence and sex ratio. While A. colemani and T. brassicae, but not C. glomerata, were able to detect GNA, this gustatory recognition had no effect on the acceptance of a GNA-sucrose solution. In all three species, GNA ingestion reduced parasitoid survival significantly. However, in respect to fecundity, negative effects were observed for T. brassicae but not for A. colemani. The results suggest that the effect of GNA consumption may depend on the specifics of a parasitoid's biology, especially its longevity and its mode of egg maturation.