Olive Fly Research Articles

Precise annotation of Drosophila mitochondrial genomes leads to insights into AT-rich regions

In the present study, we performed precise annotation of Drosophila melanogaster, D. simulans, D. grimshawi, Bactrocera oleae mitochondrial (mt) genomes using pan RNA-seq analysis. Using PacBio cDNA-seq data from D. simulans, we precisely annotated the Transcription Initiation Sites (TISs) of the mt Heavy and Light strands in Drosophila mt genomes and reported that the polyA(+) and polyA(-) motifs in the CRs are associated with TISs. The discovery of the conserved polyA(+) and polyA(-) motifs provides insights into many polyA and polyT sequences in CRs of insect mt genomes, leading to reveal the mt transcription and its regulation in invertebrates. Notably, we propose that: (1) polyA/polyT motifs in CRs function as signals to initiate mtDNA transcription; (2) the duplication, recombination or mutation of these polyA/polyT sequences formed the AT-rich regions during evolution; and (3) since CRs of many invertebrate species still contain many polyA/polyT sequences, there is a high probability that several TISs and TTSs exist in invertebrate mt genomes.

Collection and Processing of Behavioural Data of the Olive Fruit Fly, Bactrocera oleae, When Exposed to Olive Twigs Treated with Different Commercial Products

The need for the development of sustainable control methods of herbivorous insects implies that new molecules are proposed on the market. Among the different effects the new products may have on the target species, the alteration of insect oviposition behaviour might be considered. At the scope, parallel simple behavioural assays can be conducted in arena. Freely available software can be used to track observed events, but they often need intensive customization to the specific experimental design. Hence, integrating such software with, e.g., R environment, can provide a much more effective protocol development for data collection and analysis. Here we present a dataset and protocol for processing data of the oviposition behaviour of the olive fruit fly, Bactrocera oleae, when exposed to olive twigs treated with different commercial products. Treatments were rock powder, propolis, a mixture of rock powder and propolis, copper oxychloride, copper sulphate, and water as the experimental control. JWatcher was used to simultaneously collect data from 12 arena assays and ad-hoc developed R code was used to process raw data for data analyses. The procedure described here is novel and represents a valuable and transferable protocol to analyse observational events in B. oleae, as well as other biological systems.

Impacts of the olive fruit fly on the ‘Cyprus Local’ and ‘Gemlik’ olive cultivars

The objective of this study was to investigate damage rate and impact of Bactrocera oleae on olive oil quality on the ‘Cyprus Local’ and ‘Gemlik’ cultivars in Northern Cyprus. This study included 11 olive groves which were located in two different districts. A total of 11 olive groves from two separate cities (Güzelyurt and Girne) were selected during the maturity time where cvs, ‘Cyprus Local’ and ‘Gemlik’ are grown intensively. Two hundred and fifty fruits (25 fruits/10 trees) were randomly selected from each orchard and the damaged fruits were counted. These data were compared with the environmental conditions to determine relationships among the damage rates and environmental conditions.The results showed that the damage rate of olive fruit fly had a moderate correlation with the temperature, while the temperatures above 20°C were found to provoke the highest damage rate on both cultivars. Moreover, the results showed that cv. ‘Cyprus Local’ is more sensitive to olive fruit fly damage, especially at higher temperatures. An important result of the current work is that an increase in the damage rate raises the acidity ratio of the fruits and reduces the fruit quality.

Effect of Production System (Organic versus Conventional) on Olive Fruit and Oil Yields and Oil Quality Parameters in the Messara Valley, Crete, Greece; Results from a 3-Year Farm Survey

The demand for organic olive oil has increased rapidly over the last 40 years, but there is limited information on the effects of organic production methods on commercially and nutritionally relevant quality parameters in olive oil. The main objective of this farm survey-based study was therefore to compare fruit and oil yields and important oil quality parameters (including acidity, peroxide value and fatty acid profiles) between organic and conventional farms located in the Messara Plain and foothills. As expected, yields were substantially higher on farms in the Messara Plain compared with those in the foothills which have poorer soil, less access to irrigation water and are the more extensively managed. However, different to the many previous studies (which reported lower yields in organic systems), both fruit and oil yields were not significantly different in organic and conventional production and numerically ~10% higher in organic production. Additionally, olive oil quality was very high, and no substantial effects of production systems and farm location were found. Potential factors (e.g., low olive fly pressure) which may have contributed to the lack of a yield and quality gap between organic and convention production in the Messara region are discussed.

In the Spotlight-Established Researcher.

In the Spotlight-Established Researcher.

One drop only. Easy and rapid Raman evaluation of β-carotene in olive oil and its relevance as an index of olive fly attack

This paper presents, for the first time, a method for the rapid quantification of β-carotene in olive oil by Raman spectroscopy. Using a 532 nm Raman laser source, our procedure requires only one drop (100 µL) of oil, for β-carotene content to be determined. Results show that β-carotene content is associated with the lutein/β-carotene ratio, a parameter whose value describes how healthy the olives were before processing, specifically whether an olive fly attack occurred. Since olive fly attacks are not always visible to the oil producers, this method gives them the means to control the validity of the prevention strategies they adopted.

Dynamic interactions between the symbiont Candidatus Erwinia dacicola and its olive fruit fly host Bactrocera oleae

The olive fruit fly, Bactrocera oleae, the most serious pest of olives, requires the endosymbiotic bacterium Candidatus Erwinia dacicola in order to complete its development in unripe green olives. Hence, a better understanding of the symbiosis of Ca. E. dacicola and its insect host may lead to new strategies for B. oleae control. The relative abundance of bacteria during the fly life cycle comparing black and green olives was estimated by real time quantitative PCR revealing significant fluctuations during development in black olives with a peak of the bacteria in the second instar larvae. By microscopy analysis of larvae, we show that the bacteria reside extracellularly in the gastric caeca. During the transition to late third instar larvae, the bacteria were discharged into the midgut concomitant with a change in caeca size and morphology due to the contraction of the muscles surrounding the caeca. A similar alteration was also observed in a laboratory strain devoid of bacteria. To further investigate the symbiotic interaction and the change in caeca morphology a comparative transcriptomics analysis was undertaken. Samples of dissected caeca from second and third instar larvae collected from the field as well as second instar larvae from a laboratory strain devoid of symbionts showed significant changes in transcript expression. This highlighted genes associated with the developmental changes revealed by the microscopic analysis as well as responses to microorganisms.

Methodological approach to spatial analysis of agricultural pest dispersal in olive landscapes

The effectiveness of a Geographical Information Systems cost-distance tool for detecting landscape permeability in relation to the movement of pests in olive landscapes was established. The simplification of agricultural systems is linked to an increased incidence of pests on crops. Therefore, it is important to understand the impact of different land uses surrounding olive groves on pests. In this work, we analysed the effect of the structure of the olive landscape on the movement of two main olive pests—the olive fruit fly, Bactrocera oleae (Rossi) (Diptera: Tephritidae) and the olive moth, Prays oleae (Bernard) (Lepidopetera: Praydidae). We applied linear mixed effects models to analyse the relationship between pest abundance and cost-distance, using different hypotheses to evaluate those land uses that are favourable or unfavourable for the movement of these pests. The results show that this methodology is effective in detecting possible unfavourable land uses with a barrier effect, such as woodland and artificial land uses, and favourable land uses with a corridor effect such as olive groves. Whether other land uses, such as scrubland or riverbanks, act as a barrier or corridor depends on the pest and its life cycle stage. The effect that different land uses have in maintaining low levels of pest populations and ensuring the long-term sustainability of these agricultural systems are discussed. The implications of landscape permeability for the physical structure of the landscape and the dispersal of organisms, and the potential of that landscape to impact the continuous flow of natural processes are also addressed.

Identifying an Optimal Screen Mesh to Enable Augmentorium-Based Enhanced Biological Control of the Olive Fruit Fly Bactrocera oleae (Diptera: Tephritidae) and the Mediterranean Fruit Fly Ceratitis capitata (Diptera: Tephritidae).

The augmentorium is a cost-effective screened enclosure designed to receive fruits infested with insect pests, retain the pests inside but let their natural enemies escape to enhance biological control of pest populations. Screen selection is critical to ensure that an augmentorium is effective for a particular system. Here we tested five types of screens with a mini-augmentorium design and measured the escape of four insect species under laboratory conditions: the pests olive fruit fly Bactocera oleae (Rossi) (Diptera: Tephritidae) and Mediterranean fruit fly Ceratitis capitata (Wiedemann) (Diptera: Tephritidae), and the parasitoids Psyttalia lounsburyi (Silvestri) (Hymenoptera: Braconidae) and Psyttalia ponerophaga (Silvestri) (Hymenoptera: Braconidae). The sex ratio of insects that escaped the screens was compared to the sex ratio of insects that could not escape. Results showed that one screen type ('crystal mesh') was the best for the purpose of designing a functional augmentorium: it retained 90% of B. oleae adults and 100% of C. capitata adults while letting 72% of Psyttalia lounsburyi adults and 94% of P. ponerophaga adults escape. The other screen types tested were suboptimal, either because they let too many flies freely escape or because they retained too many parasitoids. Sex ratio was almost always similar for insects that managed to escape the screens and insects that were retained, except for P. ponerophaga and the screen type 'light mesh'. These results are promising for the development of a functional augmentorium against the olive fruit fly and further implementation against Mediterranean fruit fly.

Olive Fruit Fly Symbiont Population: Impact of Metamorphosis.

The current symbiotic view of the organisms also calls for new approaches in the way we perceive and manage our pest species. The olive fruit fly, the most important olive tree pest, is dependent on an obligate bacterial symbiont to its larvae development in the immature fruit. This symbiont, Candidatus (Ca.) Erwinia dacicola, is prevalent throughout the host life stages, and we have shown significant changes in its numbers due to olive fruit fly metamorphosis. The olive fruit fly microbiota was analyzed through 16S metabarcoding, at three development stages: last instar larvae, pupae, and adult. Besides Ca. E. dacicola, the olive fruit flies harbor a diverse bacterial flora of which 13 operational taxonomic units (grouped in 9 genera/species) were now determined to persist excluding at metamorphosis (Corynebacterium sp., Delftia sp., Enhydrobacter sp., Kocuria sp., Micrococcus sp., Propionibacterium sp., Pseudomonas sp., Raoultella sp., and Staphylococcus sp.). These findings open a new window of opportunities in symbiosis-based pest management.

Interaction networks between spittlebugs and vegetation types in and around olive and clementine groves of Corsica; implications for the spread of Xylella fastidiosa

The bacterium Xylella fastidiosa (Xf) is a major threat to European agriculture and notably to the olive and Citrus industry. It is transmitted by xylem-feeding insects, whose plant-feeding preferences still require investigation in European agroecosystems. Here we studied olive and clementine groves of the oriental plain of Corsica. Insect-vegetation type interaction networks for nymphs and adults of xylem-feeding insects were investigated three times a year during two years in and around crops. Networks were dominated by the Philaenus spumarius – Cistus monspeliensis interaction and were similar for olive and clementine groves, despite differences in plant communities of the ground vegetation and agricultural practices. To a lesser extent, Dittrichia viscosa was also a suitable feeding plant for P. spumarius, the main vector of Xf in Europe. Neophilaenus campestris was associated with grove ground vegetation, whereas Lepyronia coleoptrata and Aphrophora alni exhibited no or weak association with any of the target vegetation types. All species occurred on olive and clementine foliage in similar low abundance. Our results suggest that soil tillage in spring would be less efficient than in Italy to control P. spumarius. Instead, removing C. monspeliensis bushes from the close vicinity of groves could reduce risk of spread of Xf to crops. Finally, conserving D. viscosa in olive groves as a biological control strategy against the olive fruit fly is rather counter-productive as it maintains populations of P. spumarius under susceptible crops. Overall, our study shows that management strategies should be designed in relation to local insect-plant interaction networks that should be investigated in both cultivated areas and their immediate environment.

Oviposition Deterrent Activity of Fungicides and Low-Risk Substances for the Integrated Management of the Olive Fruit Fly Bactrocera oleae (Diptera, Tephritidae).

Simple SummaryThe olive fruit fly Bactrocera oleae is a very common pest infesting olive orchards wherever they are cultivated, representing the greatest threat to olive production and oil quality. Although broad-spectrum insecticides are often used to protect olive crops against B. oleae, there is increasing concern about their effects on the environment and human health. An important tool in integrated olive fly management could be the use of products with a repellency and oviposition deterrence effect. This research yielded experimental evidence of significant oviposition deterrent activity on the olive fly as side effects of substances used in olive growing such as fungicides or plant biostimulants, highlighting the potential use of these products in B. oleae management.The control of Bactrocera oleae is fundamental to decreasing the significant production loss in olive cultivation. However, traditional containment based on the use of synthetic insecticides has been encountering serious limitations due to their negative effect on human health and the environment. Within the scope of integrated olive fly management, the use of products with repellency and oviposition deterrent activity might represent a more eco-friendly solution. In this study, we tested the oviposition deterrent activity of some commercial formulations already used in olive tree crops as fungicides (copper oxychloride, dodine, mancozeb, pyraclostrobin and difeconazole) and plant bio-stimulants (tannins, clay, flavonoids and a zinc-copper-citric acid biocomplex). The trials were conducted testing the oviposition behavior of mated olive fly females in both choice and no-choice assays. Our results showed that most of the substances have affected the ovipositional activity of the olive fly, except for difeconazole. Moreover, some products (copper oxychloride, flavonoids and tannins) have proven to differently influence the flies’ oviposition comparing the two tests. The repellent effect of these commercial products should be further studied to prove whether the repellency was due either to the active ingredient or to the co-formulants, and to assess their effect in the open field.

Landscape simplification increases Bactrocera oleae abundance in olive groves: adult population dynamics in different land uses

Bactrocera oleae is the main pest in olive groves, and its management requires a sustainable perspective to reduce the use of chemical products. Landscape context is being considered as an important driver of pest reduction, but results on B. oleae show inconsistency to date. Most of landscape-pest control studies focus on the dynamics of the pests within the focal crop, ignoring these dynamics in other land uses. Here we present a study in which we analyze the seasonal population dynamics of the olive pest B. oleae in the most important land uses of a typical olive landscape in Portugal. We found that B. oleae is present in all the land uses and the dynamics are very similar to those in the olive groves. However, the presence of these land uses in the landscape did not display any increase in B. oleae abundance within the olive groves. In contrast, a landscape mainly composed by olive groves increased the abundance of this pest. Importantly, more diverse landscapes surrounding olive groves reduce the abundance of the olive fly. Based on these findings, we can conclude that B. oleae is present in all the land uses of the studied landscape but that this presence does not imply an increase of B. oleae in olive groves. Indeed, other land uses can promote landscape diversification which is a driver of the reduction of B. oleae populations in olive groves. We thus encourage olive stakeholders to increase landscape diversification around their farms by promoting/restoring other crops/habitats.

Contact Toxicity and Ovideterrent Activity of Three Essential Oil-Based Nano-Emulsions against the Olive Fruit Fly Bactrocera oleae

The control strategies for the olive crop key pest, Bactrocera oleae, involve synthetic chemical insecticides and few eco-sustainable alternatives, such as ovideterrents and lures. In the last few decades, the interest concerning the formulation of botanical based biopesticides increased, but little research investigated the suitability of these approaches for B. oleae control. This research aimed to investigate the residual contact toxicity and the oviposition deterrence of three essential oil (EO)-based nano-emulsions (Pimpinella anisum, Foeniculum vulgare, Mentha × piperita) against B. oleae adult flies. All the nano-emulsions possessed optimal physical characteristics, with droplets dimensions ranging from 115 to 152 nm and low PDI values (<0.2), even after 1 year of storage. Although no notable residual contact toxicity was noted, all the tested formulations reduced the number of oviposition puncture in no-choice tests (percent repellence: mint < fennel < anise). In choice trials, olives treated with fennel and anise EO-formulations at the highest concentration (7.5%, 75 g of EO/L) were less attractive respect to control fruits and a significant reduction of olive punctures was recorded. Nano-biopesticides are promising eco-friendly tools to integrate B. oleae pest management programs and to reduce the use of harmful conventional active ingredients.

The Infestation of Olive Fruits by Bactrocera oleae (Rossi) Modifies the Expression of Key Genes in the Biosynthesis of Volatile and Phenolic Compounds and Alters the Composition of Virgin Olive Oil.

Bactrocera oleae, the olive fruit fly, is one of the most important pests affecting the olive fruit, causing serious quantitative and qualitative damage to olive oil production. In this study, the changes induced by B. oleae infestation in the biosynthesis of volatile and phenolic compounds in olive (cvs. Picual, Manzanilla, and Hojiblanca) have been analyzed. Despite cultivar differences, the oils obtained from infested fruits showed a significant increase in the content of certain volatile compounds such as (E)-hex-2-enal, ethanol, ethyl acetate, and β-ocimene and a drastic decrease of the phenolic contents. The impact of those changes on the inferred quality of the oils has been studied. In parallel, the changes induced by the attack of the olive fly on the expression of some key genes in the biosynthesis of volatile and phenolic compounds, such as lipoxygenase, β-glucosidase, and polyphenol oxidase, have been analyzed. The strong induction of a new olive polyphenol oxidase gene (OePPO2) explains the reduction of phenolic content in the oils obtained from infested fruits and suggest the existence of a PPO-mediated oxidative defense system in olives.

Field and Laboratory Efficacy of Low-Impact Commercial Products in Preventing Olive Fruit Fly, Bactrocera oleae, Infestation.

Simple SummaryThe adoption of sustainable methods for herbivore pest control has become mandatory in Europe, with the EU directive 128/09. Since then, stringent evaluation protocols have been applied to insecticides and several molecules (that are suspected to be unsafe for the environment or human health) have been banned. Hence, the evaluation of sustainable methods, e.g., preventive tools based on the manipulation of pest behaviour, must be considered. Using field and laboratory assays, we tested the efficacy of different products in preventing infestation of a key pest of olive orchards, the olive fruit fly Bactrocera oleae. Our findings may be useful for the development of control strategies in integrated pest management (IPM) and organic agriculture.The olive fruit fly, Bactrocera oleae, is the key pest of olive trees in several areas of the world. Given the need for the development of sustainable control methods, preventive tools, based on the manipulation of pest behaviour, must be considered. Here, under field and laboratory conditions, we tested the efficacy of different products in preventing B. oleae infestation. A field trial was conducted, from July to November 2020, in an olive orchard located in Central Italy. A table olive variety was selected and sprayed with rock powder, propolis, the mixture of both, copper oxychloride, or water (control). All treatments, except propolis, caused a reduction of B. oleae oviposition in olives, compared to the control. The mixture allowed the strongest reduction of fly infestation throughout the season, suggesting a synergistic effect. Behavioural no-choice assays were conducted to better understand the effects of treatments on B. oleae females. Compared to the control, females showed a lower preference for the central area of an arena containing an olive twig bearing two olive fruits, fully developed, but still green, treated with rock powder, plus propolis mixture. For all treatments, B. oleae showed lower oviposition events, suggesting deterrence to oviposition. Our results indicate that the tested products may have value against B. oleae, within integrated pest management (IPM) and organic agriculture.

Zelus renardii Roaming in Southern Italy.

Simple SummaryThe leafhopper assassin bug Zelus renardii (Hemiptera: Reduviidae) first entered Europe in 2012 and has since acclimatised. Z. renardii is now a common insect predator in agroecosystems and urban and peri-urban areas. We performed a metadata analysis of 165 years of Z. renardii literature. Moreover, we provide laboratory tests of Z. renardii predation. The latter indicates Z. renardii interplay with relevant insect targets, such as Philaenus spumarius, Neophilaenus campestris, Bactrocera oleae, Kermes vermilio, Nidularia pulvinata, Harmonia axyridis, Apis mellifera, Aleurocanthus spiniferus, Aleurothrixus floccosus, Macrohomotoma gladiata, Drosophila suzukii, Drosophila melanogaster, Megaselia scalaris, Pseudococcidae, Miridae, and Issidae. Furthermore, predation on Aphrophoridae and other olive pests brands Z. renardii as a good mass-rearing candidate for inundative biocontrol programs of Xylella fastidiosa pauca ST53 infections and could also reduce damage caused by other olive pests. Overall, this reduviid is harmless to humans and beneficial insects.This study collects data from the literature and updates our Zelus renardii Kolenati, 1856 (Leafhopper Assassin Bug, LAB) prey knowledge. The literature consists of ca. 170 entries encompassing the years 1856 to 2021. This reduviid originated in the Nearctic region, but has entered and acclimatised in many Mediterranean countries. Our quantitative predation experiments—in the laboratory on caged plants plus field or environmental observations—confirm that LAB prefers a selected array of prey. Laboratory predation tests on living targets (Hemiptera, Coleoptera, Diptera, and Hymenoptera) agree with the literature. Zelus renardii prefers comparatively large, highly mobile, and readily available prey. LAB preferences on available hemipterans targets suggest that Zelus renardii is a good inundative biocontrol agent for Xylella fastidiosa pauca ST53 infections. LAB also prey on other important olive pests, such as Bactrocera oleae. Therefore, Zelus renardii is a major integrated pest management (IPM) component to limit Xylella fastidiosa pandemics and other pest invasions.

The estimation of optimal harvesting period for olives, based on losses in the physical and mechanical properties caused by olive fruit fly infestation at different orchard altitudes

The estimation of optimal harvesting period for olives, based on losses in the physical and mechanical properties caused by olive fruit fly infestation at different orchard altitudes

Effect of Organic Farming and Agricultural Abandonment on Beneficial Arthropod Communities Associated with Olive Groves in Western Spain: Implications for Bactrocera oleae Management

Simple SummaryOlive cultivation has been extremely relevant in the Mediterranean area for centuries, creating traditional landscapes with high cultural and biodiversity values. However, in recent decades, these landscapes have been affected by two processes. On the one hand, the most productive areas have undergone significant intensification, with greater input of agrochemicals and a much higher tree density; on the other hand, marginal areas, with lower production, are being progressively abandoned. While more attention has been paid to intensification effects, few studies have considered the consequences of olive grove abandonment. In our study, we analyzed how abandonment and management regimes (organic or traditional) affected the main olive pest (Bactrocera oleae) and different groups of natural enemies in olive groves established near the border between Spain and Portugal. Our results showed that abandoned and managed olive groves had different, but similarly rich and diverse, communities of natural enemies, highlighting the complementary role that these two habitats play at the landscape scale. Moreover, abandoned groves may not be acting as a reservoir for the olive fly. To prevent land abandonment from continuing, measures such as organic farming or agritourism, which have been implemented in the studied area, could be effective.Agricultural abandonment and intensification are among the main land-use changes in Europe. Along with these processes, different proposals have been developed to counteract the negative effects derived from agricultural intensification, including organic management. In this context, we aimed to determine how organic management and farmland abandonment affect Bactrocera oleae and its main groups of natural enemies: hymenopteran parasitoids, spiders, ants, carabids, and staphylinids. Between May and October 2018, four samplings were carried out in nine olive groves (three under organic management, three under traditional management, and three abandoned) in a rural area on the border between Spain and Portugal (Salamanca, Western Spain). Our results suggested differences between the natural enemy community composition of abandoned and organic groves, with slightly higher levels of richness and abundance in abandoned groves. We found no differences between organic and traditional groves. The managed olive groves sustained a different natural enemy community but were similarly rich and diverse compared with the more complex abandoned groves, with the latter not acting as a reservoir of B. oleae in our study area. Both systems may provide complementary habitats; however, further abandonment could cause a reduction in heterogeneity at the landscape scale and, consequently, a biodiversity loss.

Study of the interaction between the olive tree pests and their natural enemies

The aim of this investigation was to study the interaction between the olive pests and their natural enemies for the establishment of possible control programs and prevention against the main pests of olive trees and to ensure the agro-ecological stability of olive orchards and their protection. Using several sampling techniques, presence of four pests and five predators was observed on the olive tree. The phytophagous insects included serious olive tree pests; Bactrocera oleae (Rossi) (Diptera: Tephritidae), Euphyllura olivina (Costa) (Hemiptera: Psyllidae), Liothrips oleae Costa (Thysanoptera: Phlaeothripidae) and Parlatoria oleae (Colvée) (Hemiptera: Diaspididae). Significant positive correlations were acquired between the presence of the predators Chrysoperla carnea and Anthocoris nemoralis with the four pests of the olive tree. The presence of the predators, Scymnus marinus, Scymnus subvillosus and Nesidiocoris tenuis was insignificant compared to the olive pests. The activity of Hymenoptera parasitoids such as Ephedrus plagiator, Bracon sp., Encyrtidae sp. and Diglyphus sp. is necessary for the populations of B. oleae and E. Olivina.