Aphicidal Activity and Phytotoxicity of Citrus sinensis Essential-Oil-Based Nano-Insecticide.

Due to its high polyphagy, Aphis gossypii is considered a key pest of many crops, and it can feed on hundreds of plant species belonging to the families Cucurbitaceae, Malvaceae, Solanaceae, Rutaceae, and Asteraceae. The control of this pest mainly relies on synthetic insecticides whose adverse effects on the environment and human health are encouraging researchers to explore innovative, alternative solutions. In this scenario, essential oils (EOs) could play a key role in the development of ecofriendly pesticides. In this study, the development of a citrus peel EO-based nano-formulation and its biological activity against A. gossypii both in the laboratory and field were described and evaluated. The phytotoxicity towards citrus plants was also assessed. The developed nano-insecticide highlighted good aphicidal activity both in the laboratory and field trials, even at moderate EO concentrations. However, the highest tested concentrations (4 and 6% of active ingredient) revealed phytotoxic effects on the photosynthetic apparatus; the side effects need to be carefully accounted for to successfully apply this control tool in field conditions.

Abiotic and Herbivory Combined Stress in Tomato: Additive, Synergic and Antagonistic Effects and Within-Plant Phenotypic Plasticity.

BACKGROUND: Drought, N deficiency and herbivory are considered the most important stressors caused by climate change in the agro- and eco-systems and varied in space and time shaping highly dynamic and heterogeneous stressful environments. This study aims to evaluate the tomato morpho-physiological and metabolic responses to combined abiotic and herbivory at different within-plant spatial levels and temporal scales. METHODS: Leaf-level morphological, gas exchange traits and volatile organic compounds (VOCs) profiles were measured in tomato plants exposed to N deficiency and drought, Tuta absoluta larvae and their combination. Additive, synergistic or antagonistic effects of the single stress when combined were also evaluated. Morpho-physiological traits and VOCs profile were also measured on leaves located at three different positions along the shoot axes. RESULTS: The combination of the abiotic and biotic stress has been more harmful than single stress with antagonistic and synergistic but non-additive effects for the morpho-physiological and VOCs tomato responses, respectively. Combined stress also determined a high within-plant phenotypic plasticity of the morpho-physiological responses. CONCLUSIONS: These results suggested that the combined stress in tomato determined a "new stress state" and a higher within-plant phenotypic plasticity which could permit an efficient use of the growth and defense resources in the heterogeneous and multiple stressful environmental conditions.

Toxics or Lures? Biological and Behavioral Effects of Plant Essential Oils on Tephritidae Fruit Flies.

The family Tephritidae (Diptera) includes species that are highly invasive and harmful to crops. Due to globalization, international trade, and human displacement, their spread is continuously increasing. Unfortunately, the control of tephritid flies is still closely linked to the use of synthetic insecticides, which are responsible for detrimental effects on the environment and human health. Recently, research is looking for alternative and more eco-friendly tools to be adopted in Integrated Pest Management (IPM) programs. In this regard, essential oils (EOs) and their main compounds represent a promising alternative to chemical insecticides. EOs are made up of phytoconstituents formed from the secondary metabolism of many plants and can act as attractants or toxics, depending on the dose. Because of this unique characteristic, EOs and their main constituents are promising tools that can be used both in Sterile Insect Technique (SIT) programs and in the "lure and kill" technique, exploiting the attractiveness of the product in the former case and its toxicity in the latter. In this article, current knowledge on the biological and behavioral effects of EOs and their main constituents on tephritid fruit flies is reviewed, mainly focusing on species belonging to the Anastrepha, Bactrocera, Ceratitis, and Zeugodacus genera. The mechanisms of action of EOs, their real-world applications, and challenges related to their use in IPM are critically discussed.

Volatile Infochemicals from Rhyzopertha dominica Larvae and Larval Feces Involved in Theocolax elegans Host Habitat Location.

The development of biologically based approaches for stored product pest control is needed to reduce chemical inputs. Bioassays were performed to investigate host habitat location in the trophic interaction durum wheat/Rhyzopertha dominica/Theocolax elegans. GC-MS analyses were carried out to identify some chemical compounds produced by the host-related substrates. Choice and no-choice experiments demonstrated that female parasitoids were poorly attracted to intact kernels with respect to the infested substrates. Furthermore, T. elegans females performed longer residence time on infested wheat, and they generally displayed a short-term like fidelity for this cue, remaining on it during the whole observation. Infested wheat emitted one chemical (fenchone), which is possibly linked to host larvae presence, while the feces produced by host larvae emitted some other characteristic compounds. Results demonstrated that the presence of host larvae is a key factor for T. elegans host habitat location, considering that the attractiveness of the undamaged wheat is irrelevant. Furthermore, parasitoid females tended to stay on attractive cues, limiting the risk of contamination of other commodities. Biological control tools may be useful to improve the efficiency of pest management programs, but their application should be carefully evaluated.

Side effects of two citrus essential oil formulations on a generalist insect predator, plant and soil enzymatic activities.

The widespread use of chemical pesticides for crop protection, despite having contributed to ensure food security, have shown to exert negative impacts on the environment and on human health. In addition, the frequent emergence of resistance to pesticides and their adverse effects toward non-target organisms have generated the need to develop novel ecofriendly tools for pest control. Among these, plant essential oils (EOs) may play a central role in arthropod pest control. Recently, two formulations (Emulsion and PEG-nanoparticles) of three citrus EOs (lemon, mandarin and sweet orange) showed a promising potential against Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae), a key tomato pest. Here, we evaluated the side effects of these experimental insecticides active substances toward (i) the generalist predator of several tomato pests, Nesidiocoris tenuis Reuter (Hemiptera: Miridae); (ii) the soil enzymatic activities (dehydrogenase activity, alkaline phosphomonoesterase, acid phosphomonoesterase and urease) and (iii) the tomato plant antioxidant enzymes (ascorbate peroxidase, catalase, superoxide dismutase and polyphenol oxidase). Among the tested formulations, mandarin EO-based insecticide presented a significant impact on the predator survival and reproduction. Conversely, all the tested compounds proved to be harmless for the soil enzymatic and the plant antioxidant activities. Overall, these results provide solid bases for the development of novel biopesticides for sustainable tomato crop protection.

Olive fruit volatiles route intraspecific interactions and chemotaxis in Bactrocera oleae (Rossi) (Diptera: Tephritidae) females.

Plant nutritional quality and chemical characteristics may affect the fitness of phytophagous insects. Here, the olfactory preferences of Bactrocera oleae (Rossi) females toward olives with different maturation and infestation status were evaluated in three cultivars: Ottobratica, Roggianella and Sinopolese. Volatile profiles from olives were identified by SPME/GC-MS. Choice tests were performed to determine the responses of B. oleae adult females toward fruits and pure chemicals linked to infestation degree. Cultivar was the main source of variability explaining the differences recorded in volatile emissions. Moreover, three VOCs [ß-myrcene, limonene and (E)-ß-ocimene] were associated to infestation status across all olive varieties. In choice-tests, B. oleae females always preferred the olfactory cues from low-infested over high-infested fruits. Therefore, choice-tests using synthetic VOCs, emitted in greater amount by high-infested fruit, were arranged in order to identify putative B. oleae kairomones. While females were indifferent to ß-myrcene, the highest dosages of limonene and (E)-ß-ocimene were unfavoured by the tested flies, which preferentially moved toward the empty arm of the Y-tube. Furthermore, females preferred the lowest concentration of ß-ocimene compared to the highest one. These results supported our hypothesis that fruit VOCs may serve as kairomones for female flies.

Author Correction: Citrus peel essential oil nanoformulations to control the tomato borer, Tuta absoluta: chemical properties and biological activity.

A correction to this article has been published and is linked from the HTML and PDF versions of this paper. The error has been fixed in the paper.

Instar- and host-associated differentiation of bacterial communities in the Mediterranean fruit fly Ceratitis capitata.

Microorganisms are acknowledged for their role in shaping insects' evolution, life history and ecology. Previous studies have shown that microbial communities harbored within insects vary through ontogenetic development and among insects feeding on different host-plant species. In this study, we characterized the bacterial microbiota of the highly polyphagous Mediterranean fruit fly, Ceratitis capitata (Diptera: Tephritidae), at different instars and when feeding on different host-plant species. Our results show that the bacterial microbiota hosted within the Mediterranean fruit fly differs among instars and host-plant species. Most of the bacteria harbored by the Mediterranean fruit fly belong to the phylum Proteobacteria, including genera of Alphaproteobacteria such as Acetobacter and Gluconobacter; Betaprotobacteria such as Burkholderia and Gammaproteobacteria such as Pseudomonas.

VOC emissions influence intra- and interspecific interactions among stored-product Coleoptera in paddy rice.

Olfaction is a pivotal sense for insects and granivorous pests may exploit grain volatiles for food selection. Tribolium confusum, is a secondary pest of stored cereals that benefits from primary pests' infestation, as other secondary feeders, triggering competition. This study aimed to evaluate the preferences of T. confusum females toward different-infested paddy rice, highlighting the impact of intra- and interspecific competition. Tribolium confusum showed positive chemotaxis toward rice infested by larvae of a primary pest (Sitophilus zeamais), but not for grain attacked by adults alone. Furthermore, kernels concurrently infested by a primary (S. zeamais) and a secondary pest (T. confusum or Cryptolestes ferrugineus) were evaluated in Y-tube bioassays, highlighting that both food-sources were innately attractive for T. confusum females. Moreover, females positively oriented toward rice infested by conspecifics, while they avoided grain infested by C. ferrugineus, averting an extremely competitive habitat. Behavioural responses of T. confusum females and volatile emissions of different-infested rice highlighted the occurrence of plant-mediated interactions among insects from the same trophic guild. Seventy volatiles were identified and significant differences among the tested food-sources were recorded, emphasizing the presence of 6 putative attractants and 6 repellents, which may be useful biocontrol tools.

Influence of Host Plant on Thaumetopoea pityocampa Gut Bacterial Community.

Microbial communities associated to the gut of insects are attracting an increasing interest, mainly because of their role in influencing several host life-traits. The characterization of the gut microbial community is pivotal for understanding insect ecology and, thus, to develop novel pest management strategies. The pine processionary moth, Thaumetopoea pytiocampa (Denis & Schiff.) (Lepidoptera: Thaumetopoeidae), is a severe defoliator of pine forests, able to feed on several pine species. In this work, we performed a metabarcoding analysis to investigate, for the first time, the diversity of the gut bacterial community of pine processionary larvae associated with three different host pine species (Pinus halepensis, Pinus nigra subsp. laricio, and Pinus pinaster). We found that the gut microbial community of T. pityocampa larvae collected on P. halapensis was different from that associated with larvae collected from P. nigra and P. pinaster. Moreover, the high presence of bacteria belonging to the genera Modestobacter, Delftia, and unidentified Methylobacteriaceae retrieved in larvae feeding on P. halapensis suggested that specific interactions can occur. Our results provide the evidence that different host plant differently impact on the microbiota diversity of T. pityocampa larvae, contributing to the general knowledge of this pest with information that could be useful in shaping the next generation of pest control strategies.

Citrus peel essential oil nanoformulations to control the tomato borer, Tuta absoluta: chemical properties and biological activity.

The repeated use of conventional synthetic pesticides in crop protection leads to resistance development by pests along with a negative impact on the environment, particularly non-target arthropods. Plant-derived active compounds, such as essential oils (EOs), play a key role in sustainably controlling pests. The lethal and sublethal activity of citrus peel EOs as emulsions and included in polyethylene glycol (PEG) nanoparticles (EO-NPs) was determined against the invasive tomato pest Tuta absoluta. Their effects on the plants were also assessed. The results showed an overall good insecticidal activity of the compounds tested, with a higher mortality through contact on eggs and larvae by EO emulsions and through ingestion on larvae by EO-NPs. The nanoformulation also significantly reduced the visible toxic effects on the plants. The data collected suggest that these natural compounds, especially when nanoformulated, could be successfully used in integrated pest management programs for T. absoluta.

A novel GIS-based approach to assess beekeeping suitability of Mediterranean lands.

Honeybees are critically important for the environment and to the economy. However, there are in substantial decline worldwide, leading to serious threat to the stability and yield of food crops. Beekeeping is of pivotal importance, combining the wide economical aspect of honey production and the important ecological services provided by honeybees. In this scenario, the prompt identification of beekeeping areas is strategic, since it maximised productivity and lowered the risks of colony losses. Fuzzy logic is an ideal approach for problem-solving tasks, as it is specifically designed to manage problems with a high degree of uncertainty. This research tested a novel GIS-based approach to assess beekeeping suitability of lands located in Calabria (Southern Italy), without relying to Analytic Hierarchy Process - Multiple Criteria Decision Making (AHP-MCDM), thus avoiding the constraints due to the technique and decision makers' influences. Furthermore, the data used here were completely retrieved from open access sources, highlighting that our approach is characterized by low costs and can be easily reproduced for a wide arrays of geographical contexts. Notably, the results obtained by our experiments were validated by the actual beekeeping reality. Besides beekeeping, the use of this system could not only be applied in beekeeping land suitability evaluations, but may be successfully extended to other types of land suitability evaluations.

A Metabarcoding Survey on the Fungal Microbiota Associated to the Olive Fruit Fly.

The occurrence of interaction between insects and fungi is interesting from an ecological point of view, particularly when these interactions involve insect pests and plant pathogens within an agroecosystem. In this study, we aimed to perform an accurate analysis on the fungal microbiota associated to Bactrocera oleae (Rossi) through a metabarcoding approach based on 454 pyrosequencing. From this analysis, we retrieved 43,549 reads that clustered into 128 operational taxonomic units (OTUs), of which 29 resulted in the "core" associate fungi of B. oleae. This fungal community was mainly represented by sooty mould fungi, such as Cladosporium spp., Alternaria spp. and Aureobasidium spp., by plant pathogens like Colletotrichum spp. and Pseudocercospora spp., along with several other less abundant taxa whose ecology is unclear in most of the cases. Our findings lead to new insights into the microbial ecology of this specific ecological niche, enabling the understanding of a complex network of interactions within the olive agroecosystem.

Larvicidal Effects of Four Citrus Peel Essential Oils Against the Arbovirus Vector Aedes albopictus (Diptera: Culicidae).

In this study, we evaluated the larvicidal activity of four citrus essential oils (EOs; sweet orange, mandarin, bergamot, and lemon) against the arbovirus vector Aedes albopictus (Skuse) (Diptera: Culicidae) under laboratory conditions. Through gas chromatography-mass spectrometry (GC-MS) analyses, we found that in sweet orange, mandarin, and lemon EOs, limonene was the most abundant compound, whereas linalyl acetate was the most abundant in the bergamot EO. All tested EOs showed a marked larvicidal activity, in particular sweet orange, lemon, and bergamot that killed all treated larvae. After 24 h of exposure, the LC50 values of the tested citrus EOs ranged from 145.27 (lemon EO) to 318.07 mg liter(-1) (mandarin EO), while LC(95) ranged from 295.13 to 832.44 mg liter(-1). After 48 h of exposure, the estimated LC(50) values decreased to values ranging from 117.29 to 209.38 mg liter(-1), while LC(95) ranged from 231.85 to 537.36 mg liter(-1). The results obtained from these evaluations, together with the large availability at reasonable costs of citrus EOs, are promising for the potential development of a new botanical mosquitocide.

Life stage-dependent susceptibility of Aphytis melinus DeBach (Hymenoptera: Aphelinidae) to two pesticides commonly used in citrus orchards.

The assessment of pesticides compatibility with natural enemies is recommended before including agrochemicals in integrated and organic pest management schemes. The lethal and sublethal effects of a mineral oil and a juvenile hormone mimic (pyriproxyfen), on adults and larvae of Aphytis melinus, a key ectoparasitoid of armored scale insect pests of citrus, such as Aonidiella aurantii, were evaluated. Mineral oil caused very high mortality on the adults, while a lower acute toxicity was recorded on young instars. No significant effects on their reproduction capacity and on the sex-ratio of the progeny were observed. Pyriproxyfen had neither lethal nor sublethal effects (in terms of survived female fertility) on A. melinus adults. However, parasitoid larvae exposed to this insecticide suffered strong acute toxicity and fertility reduction (progeny number and proportion of female progeny). When adults were offered the choice to parasitize treated and untreated scales they significantly preferred the control ones, and when they were exposed to only treated scaled the parasitism rate was significantly lower only with mineral oil-treated hosts. The significant differences in the susceptibility of the two parasitoid instars highlight the importance of including this aspect in pesticide risk assessment procedures and in the choice of the pesticide and of the treatment timing in the field. Overall, the results indicate potential for integrating A. melinus, both naturally present and artificially released, and these insecticides only by appropriate timing of insecticide spraying and parasitoid releases.