The Effects of Natural Insecticides on the Green Peach Aphid Myzus persicae (Sulzer) and Its Natural Enemies Propylea quatuordecimpunctata (L.) and Aphidius colemani Viereck.

Botanical insecticides and soaps are frequently proposed as environmentally safer alternatives to synthetic insecticides. However, the efficacy and selectivity of these products are often only partially supported by empirical evidence. Here, we tested the effectiveness of five botanical insecticides, belonging to different categories, on the green peach aphid Myzus persicae (Sulzer) and their selectivity towards two natural enemies, the ladybird beetle Propylea quatuordecimpunctata (L.) and the parasitoid Aphidius colemani (Dalman). White thyme essential oil (EO), sweet orange EO, crude garlic extract and Marseille soap were tested and compared with a pyrethrin-based commercial product. Both direct spray assays and residual contact assays on treated cabbage leaf disks were carried out. The tested products had low efficacy against aphids when compared to pyrethrins but were in general less detrimental to ladybird beetle larvae, meaning that if applied against other pests, they have a lower chance of harming this agent of aphid biocontrol. Some of the products (soap, orange EO) did, however, show direct exposure toxicity toward ladybird larvae, and thyme EO had extensive phytotoxic effects on cabbage leaves, possibly indirectly leading to higher mortality in ladybird adults. These results underline the necessity for case-by-case evaluations of botanical insecticides.

Effects of three chitin synthesis inhibitors on egg masses, nymphs and adults of Halyomorpha halys (Hemiptera: Pentatomidae).

BACKGROUND: Halyomorpha halys, (brown marmorated stink bug, BMSB), is a high-concern invasive species causing severe damage to orchards in many countries outside its native Asian range. Management methods other than frequent sprays of broad-spectrum insecticides are needed to restore integrated pest management (IPM) practices in orchards. Chitin synthesis inhibitors are usually regarded as desirable options in IPM programs because of lower toxicity to beneficial insects and non-target organisms compared to neurotoxic insecticides. In this study, the activity of three chitin synthesis inhibitors (namely buprofezin, novaluron and triflumuron) was investigated on BMSB egg masses, third instars and adults by means of laboratory bioassays. RESULTS: Novaluron and to a lesser extent triflumuron were detrimental to BMSB nymphs exposed to residues on potted peach plants. Novaluron caused high mortality among early instars that emerged from sprayed egg masses. No significant differences were found between buprofezin and water control on eggs or third instars. When sprayed on BMSB adults, none of the chitin synthesis inhibitors affected survival, fecundity, or egg hatching. CONCLUSION: Given the activity on nymphs, but the lack of effects on adults, novaluron and triflumuron might be considered for field applications only as a tool in a wider management strategy along with other methods aimed at preventing the invasion of crops by BMSB adults. © 2023 Society of Chemical Industry.

Implementing Mass Rearing of Trissolcus japonicus (Hymenoptera: Scelionidae) on Cold-Stored Host Eggs.

Halyomorpha halys (Stål) (Hemiptera: Pentatomidae), a pest of Asian origin, has been causing severe damage to Italian agriculture. The application of classical biological control by the release of Trissolcus japonicus (Ashmead) (Hymenoptera: Scelionidae), an exotic egg parasitoid, appears to be one promising solution. In Italy, releases of T. japonicus in the field were authorized in 2020. In this study, some parameters that could influence the rearing of T. japonicus in insectaries were investigated. A significantly higher production of progeny was observed on host eggs stored at 6 °C (86.5%) compared to -24 °C (48.8%) for up to two months prior to exposure to parasitism. There were no significant differences in progeny production from single females in a vial provided with only one egg mass (83.2%) or 10 females inside a cage with 6 egg masses (83.9%). The exposure of parasitoids to refrigerated (6 °C) egg masses of H. halys for 72 h led to a significantly higher production of progeny (62.1%) compared to shorter exposures for 48 (44.0%) or 24 h (37.1%). A decline in production of progeny by the same female was detected between the first (62.1%) and the second parasitization (41.3%). Adult parasitoids stored at 16 °C for up to 90 days had an 87.1% survival rate, but a significant decrease in progeny production was detected. These parameters could be adjusted when rearing T. japonicus for specific aims such as the production of individuals for field release or colony maintenance.

Halyomorpha halys (Hemiptera: Pentatomidae) on Kiwifruit in Northern Italy: Phenology, Infestation, and Natural Enemies Assessment.

The brown marmorated stink bug (BMSB), Halyomorpha halys (Stål) (Hemiptera: Pentatomidae), is an Asian invasive plant-feeding insect and an emerging kiwifruit pest. Knowledge about the BMSB dynamics and damage to kiwifruit outside the bug native range is scarce. This 2-yr study was aimed at describing phenology and infestation dynamics of BMSB in green-fleshed and yellow-fleshed kiwifruits. Natural enemies were investigated as well. Field surveys were performed weekly in two sites in Northern Italy by pheromone-baited traps and visual samplings, from early Spring to mid-Autumn. All BMSB life stages were collected and kept under observation to assess occurrence of parasitoids. A sample of fruit was dissected weekly to detect BMSB feeding injuries. In 2018-2019, BMSB was found throughout the growing season with two generations per year with a large overlapping of all life stages. Pheromone-baited traps and visual samplings gave consistent results and can be both considered effective sampling methods on kiwifruit. Fruit injuries were significantly related to kiwifruit development stage, weekly captures of BMSBs by traps, and cultivar. Several BMSB egg masses were found on kiwifruit leaves. Anastatus bifasciatus Geoffroy (Hymenoptera: Eupelmidae), a native egg parasitoid with Palearctic distribution, emerged from 20.14% of eggs in 2018, but the percent parasitism dropped to 0.47% in 2019. In both years, other natural enemies were found exerting negligible pressure on BMSB populations. The determination of reliable economic thresholds for BMSB on kiwifruit is urgently needed to develop a robust and sustainable integrated pest management (IPM) strategy, and this study provides data towards that direction.

Impact of Triflumuron on Halyomorpha halys (Hemiptera: Pentatomidae): Laboratory and Field Studies.

Halyomorpha halys, (the brown marmorated stink bug, BMSB), is a high-concern invasive species causing severe damage to orchards in many countries outside its native Asian range. Control options matching both effectiveness and sustainability are currently lacking. Inhibitors of chitin biosynthesis might be exploited for integrated management programs because of the overall better ecotoxicological profile in comparison with most neurotoxic insecticides used so far against BMSB. In this study, the activity of triflumuron, a benzoylphenyl urea hampering chitin biosynthesis, was tested on BMSB in laboratory and field conditions. In laboratory bioassays, the insecticide was sprayed on potted peach plants (30 cm high) and residues were aged in a glasshouse for 0, 7, 14, and 21 d. Then, third-instar bugs were placed on the plants and continuously exposed to residues. Mortality was scored after 7, 14, and 21 d exposure. Triflumuron caused significantly higher mortality on BMSB nymphs in comparison with water controls at all aging periods. Moreover, aging of residues up to 21 d did not cause any significant reduction of activity. Field experiments were also carried out in 2019 in eight pear orchards. Injuries to fruits at harvest were compared between plots where triflumuron was added to insecticide sprays against BMSB and control plots managed exactly in the same way but without any triflumuron treatment. An overall mean of 9.99 ± 1.98% stink bug injured fruits was detected in plots managed with the strategy including triflumuron, whereas 19.45 ± 3.55% of fruits were injured in plots assigned to controls.

Use of Sugar Dispensers to Disrupt Ant Attendance and Improve Biological Control of Mealybugs in Vineyard.

Planococcus ficus (Signoret) and Pseudococcus comstocki (Kuwana) (Hemiptera: Pseudococcidae) are economically important pests occurring in vineyards, causing severe economic losses for growers and compromising bunch production. The partial effectiveness of insecticides used in controlling mealybug infestations as well as their high impact on the environment and on human health have led to the research of alternative and sustainable control methods, including biological control. Several natural enemies are reported to be effective against mealybugs, but their activity may be hindered by tending ants. These social insects are known to exhibit a mutualistic relationship with mealybugs, resulting in extremely aggressive behavior against beneficial insects. Consequently, this study explored a method to mitigate ant attendance by means of sugar dispensers in order to improve ecosystem services, as well as decrease mealybug infestation in vineyards. Field trials were carried out in four commercial vineyards of Northern Italy infested by mealybugs, in which Anagyrus vladimiri Triapitsyn (Hymenoptera: Encyrtidae) and Cryptolaemus montrouzieri Mulsant (Coleoptera: Coccinellidae) were released as biological control agents. Our results showed that sugar dispensers reduced ant activity and mealybug infestation, leading to a significant enhancement of ecosystem services. The technique showed a great potential in boosting biological control against mealybugs in field conditions, though the field application seemed to be labour intensive and needs to be replicated for a multi-year evaluation.

Influence of Cover Crop Termination on Ground Dwelling Arthropods in Organic Vegetable Systems.

A key aspect in cover crop management is termination before the cash crop is planted. The aim of this study was to assess the effects of termination methods on ground-dwelling arthropods. The conventional mechanical termination method-i.e., green manuring by means of a disc harrow-was compared to flattening using a roller crimper. Two different crop systems were investigated for two growing seasons; cauliflower was grown in autumn after the termination of a mixture of cowpea, pearl millet, and radish, and tomato was cropped in spring and summer after the termination of a mixture of barley and vetch. Ground beetles (Coleoptera: Carabidae), rove beetles (Coleoptera: Staphylinidae), and spiders (Araneae) were sampled by means of standard pitfall traps throughout the growing season of both cash crops. The roller crimper increased the overall abundance of ground beetles in the first growing season of both cash crops, whereas in the second year, no significant effect could be detected. Rove beetles were more abundant in plots where the cover crops were terminated by the roller crimper. Finally, green manuring increased the abundance of spiders, especially on the first sampling date after cover crop termination. Albeit different taxa showed different responses, the termination of cover crops by a roller crimper generally increased the abundance of ground dwelling arthropods. Given that most of the sampled species were generalist predators, their increased abundance could possibly improve biological control.

Mating Disruption of Helicoverpa armigera (Lepidoptera: Noctuidae) on Processing Tomato: First Applications in Northern Italy.

Helicoverpa armigera is a polyphagous and globally distributed pest. In Italy, this species causes severe damage on processing tomato. We compared the efficacy of mating disruption with a standard integrated pest management strategy (IPM) in a two-year experiment carried out in Northern Italy. Mating disruption registered a very high suppression of male captures (>95%) in both growing seasons. Geostatistical analysis of trap catches was shown to be a useful tool to estimate the efficacy of the technique through representation of the spatial pattern of captures. Lower fruit damage was recorded in mating disruption than in the untreated control plots, with a variable efficacy depending on season and sampling date. Mating disruption showed a higher efficacy than standard IPM in controlling H. armigera infestation in the second season experiment. Mating disruption showed the potential to optimize the H. armigera control. Geostatistical maps were suitable to draw the pheromone drift out of the pheromone-treated area in order to evaluate the efficacy of the technique and to detect the weak points in a pheromone treated field. Mating disruption and standard IPM against H. armigera were demonstrated to be only partially effective in comparison with the untreated plots because both strategies were not able to fully avoid fruit damage.

Evaluation of the potential exposure of butterflies to genetically modified maize pollen in protected areas in Italy.

Environmental impacts of genetically modified crops are mandatorily assessed during their premarket phase. One of the areas of concern is the possible impact on nontarget organisms. Crops expressing Cry toxins might affect Lepidoptera larvae living outside cultivated fields, through pollen deposition on wild plants, which constitute their food source. While pollen toxicity varies among different events, possible exposure of nontarget species depends on the agro-environmental conditions. This study was conducted in two protected areas in Italy, characterized by different climatic conditions, where many Lepidoptera species thrive in proximity to maize cultivations. To estimate the possible exposure in absence of the actual stressor (e.g., Cry1-expressing maize plants), we conducted a two-year field survey of butterflies and weeds. Indicator species were selected-Aglais (Inachis) io in the Northern site and Vanessa cardui in the Southern site-and their phenology was investigated. Pollen dispersal from maize fields was measured by collection in Petri dishes. Duration and frequency of exposure was defined by the overlap between pollen emission and presence of larvae on host plants. Different risk scenarios are expected in the two regions: highest exposure is foreseen for A. io in the Northern site, while minimal exposure is estimated for V. cardui in the Southern site. In the latter case, locally grown maize cultivars flower in mid-summer in coincidence with an aestivation period for several butterfly species due to hot and dry conditions. Moreover, host plants of V. cardui are at the end of their life cycle thus limiting food availability.

Lethal and Sub-lethal Effects of Four Insecticides on the Aphidophagous Coccinellid Adalia bipunctata (Coleoptera: Coccinellidae).

Conventional insecticide assays, which measure the effects of insecticide exposure on short-term mortality, overlook important traits, including persistence of toxicity or sub-lethal effects. Therefore, such approaches are especially inadequate for prediction of the overall impact of insecticides on beneficial arthropods. In this study, the side effects of four modern insecticides (chlorantraniliprole, emamectin benzoate, spinosad, and spirotetramat) on Adalia bipunctata (L.) (Coleoptera: Coccinellidae) were evaluated under laboratory conditions by exposition on treated potted plants. In addition to investigation of acute toxicity and persistence of harmful activity in both larvae and adults of A. bipunctata, demographic parameters were evaluated, to provide a comprehensive picture of the nontarget effects of these products. Field doses of the four insecticides caused detrimental effects to A. bipunctata; but in different ways. Overall, spinosad showed the best toxicological profile among the products tested. Emamectin benzoate could be considered a low-risk insecticide, but had high persistence. Chlorantraniliprole exhibited lethal effects on early instar larvae and adults, along with a long-lasting activity, instead spirotetramat showed a low impact on larval and adult mortality and can be considered a short-lived insecticide. However, demographic analysis demonstrated that chlorantraniliprole and spirotetramat caused sub-lethal effects. Our findings highlight that sole assessment of mortality can lead to underestimation of the full impact of pesticides on nontarget insects. Demographic analysis was demonstrated to be a sensitive method for detection of the sub-lethal effects of insecticides on A. bipunctata, and this approach should be considered for evaluation of insecticide selectivity.

The Influence of Vegetation and Landscape Structural Connectivity on Butterflies (Lepidoptera: Papilionoidea and Hesperiidae), Carabids (Coleoptera: Carabidae), Syrphids (Diptera: Syrphidae), and Sawflies (Hymenoptera: Symphyta) in Northern Italy Farmland.

Landscape structure as well as local vegetation influence biodiversity in agroecosystems. A study was performed to evaluate the effect of floristic diversity, vegetation patterns, and landscape structural connectivity on butterflies (Lepidoptera: Papilionoidea and Hesperiidae), carabids (Coleoptera: Carabidae), syrphids (Diptera: Syrphidae), and sawflies (Hymenoptera: Symphyta). Vegetation analysis and insect samplings were carried out in nine sites within an intensively farmed landscape in northern Italy. Plant species richness and the percentage of tree, shrub, and herb cover were determined by means of the phytosociological method of Braun-Blanquet. Landscape structural connectivity was measured as the total length of hedgerow network (LHN) in a radius of 500 m around the center of each sampling transect. Butterflies species richness and abundance were positively associated both to herb cover and to plant species richness, but responded negatively to tree and shrub cover. Shrub cover was strictly correlated to both species richness and activity density of carabids. The species richness of syrphids was positively influenced by herb cover and plant richness, whereas their abundance was dependent on ligneous vegetation and LHN. Rarefaction analysis revealed that sawfly sampling was not robust and no relationship could be drawn with either vegetation parameters or structural connectivity. The specific responses of each insect group to the environmental factors should be considered in order to refine and optimize landscape management interventions targeting specific conservation endpoints.

Larvicidal activity and influence of Bacillus thuringiensis var. israelensis on Aedes albopictus oviposition in ovitraps during a two-week check interval protocol.

Toxicity persistence of Bacillus thuringiensis var. israelensis (Bti) was evaluated in laboratory and field trials to develop a new protocol for Aedes albopictus monitoring. In the laboratory, the residual toxicity of 5 Bti formulations was compared using the percentage mortality at days 4, 8, 12, and 16 of Culex pipiens larvae exposed for 24 h to Bti treatments. The results showed a good performance of all tested formulations (>97% mortality at day 14, for all the formulations), but only Vectobac 12AS at the concentration of 1 ml/liter showed an efficacy of 100% for 2 wk. A field study was designed to test the effect of Bti on the ovitrap check interval or influence of ovipositional response of gravid Ae. albopictus females. Three different ovitrap treatments were used: ovitraps with tap water checked weekly; ovitraps with tap water checked every 2 wk; ovitraps with Bti (Vectobac 12AS, dose of 1 ml/liter) checked every 2 wk. Our study demonstrated that in the ovitrap, the toxic action of a 1% solution of Bti was maintained for at least 14 days with mortality of 100% and that rainfall did not seem to negatively influence the residual action of Bti. Therefore the probability that the larvae may complete the developmental cycle in ovitraps with Bti seems to be very low. The oviposition activity index showed that Bti enhances the oviposition rate of Ae. albopictus by 17.4%.

Mosquito-based survey for the detection of flaviviruses and filarial nematodes in Aedes albopictus and other anthropophilic mosquitoes collected in northern Italy.

In order to optimize resources, an assay allowing the detection of several etiological agents would be useful. This paper describes a polymerase chain reaction (PCR)-based assay for the parallel detection of different pathogens in mosquitoes (Diptera: Culicidae). The assay combines the analysis of both RNA and DNA, hence allowing the detection of both flaviviruses (Flaviviridae: Flavivirus) and filarial nematodes (Spirurida: Onchocercidae). The sensitivity of diagnostic assay for both flavivirus and nematode infections was tested by means of spiked samples and resulted adequate for the purpose of screening mosquito samples. The assay was implemented to check pathogen infections in 637 mosquitoes field-collected in Romagna, northern Italy. The integrity of RNA extracted from mosquito pools was checked by control amplifications targeting the 18 S rRNA of mosquitoes. Control amplifications were successful in 118 out of 119 pools. Flavivirus RNA was not found in any of these 118 pools, whereas DNA of Dirofilaria immitis (Leidy) was detected in one pool of Aedes albopictus (Skuse).

Parasitic hymenoptera fauna on agromyzidae (diptera) colonizing weeds in ecological compensation areas in northern Italian agroecosystems.

Parasitoids (Hymenoptera) associated with agromyzid leafminers (Diptera: Agromyzidae) were studied in three rural farms located in northern Italy. The parasitoids were reared from mined foliage of weeds growing in field margins. We reared 998 Hymenoptera specimens, representing five families, 23 genera, and 53 species, from leafminers infesting weeds. Eulophidae was the most abundant family (67.64%), followed by Braconidae (28.86%), Eucoilinae (1.40%), Tetracampidae (1.40%), and Pteromalidae (0.7%). Braconids was the most species rich family, accounting for 28 species; eulophids were represented by 19 species, pteromalids by four species, and eucoilins and tetracampids by one species each. The dominant parasitoid was the eulophid Pediobius metallicus (Nees), representing 18.17% of the total, followed by Diglyphus isaea (Walker) (12.73%), and Neochrysocharis formosa (Westwood) (10.82%). The most abundant braconid parasitoid was Dacnusa maculipes Thomson (9.62%). More than 80% of parasitoids were recovered from 10 plant species: Cirsium arvense (L.) Scopoli, Plantago lanceolata L., Sonchus asper (L.) Hill, Papaver rhoeas L., Picris echioides L., Lactuca serriola L., Myagrum perfoliatum L., Ranunculus velutinus Tenore, Arctium lappa L., and Medicago sativa L. The retention and the management of wild plants within field margins can be crucial tools to enhance the populations of biological control agents of agromyzids and to conserve rare parasitic wasp species.

Polymerase chain reaction-restriction fragment length polymorphism assays to distinguish Liriomyza huidobrensis (Diptera: Agromyzidae) from associated species on lettuce cropping systems in Italy.

The pea leafminer, Liriomyza huidobrensis (Blanchard) (Diptera: Agromyzidae), is a serious insect pest infesting open field lettuce plantings in northern Italy. In these cropping systems, it coexists with several other agromyzid species that have negligible economic importance on open field vegetables. The rapid detection of L. huidobrensis is crucial for effective management strategies, but the identification of agromyzids to species can be very difficult at adult as well at immature stages. In this study, a polymerase chain reaction (PCR)-restriction fragment length polymorphism assay is proposed to separate L. huidobrensis from Liriomyza bryoniae (Kaltenbach), Liriomyza trifolii (Burgess), and Chromatomyia horticola (Goureau), which usually occur in the same lettuce plantings. An approximately 1,031-bp region of the mitochondrial genome encompassing the 3' region of cytochrome oxidase I, the whole leucine tRNA, and all of the cytochrome oxidase II was amplified by PCR and digested using the enzymes PvuII and SnaBI separately. Both endonucleases cut the amplicons of L. huidobrensis in two fragments, whereas the original band was not cleaved in the other analyzed species. The presence of Dacnusa spp. DNA does not bias the assay, because the PCR conditions and the primer set here described do not amplify any tract of this endoparasitic wasp genome.