The Cuticular Hydrocarbons of Dasineura Oleae Show Differences Between Sex, Adult Age and Mating Status.

In insects, cuticular lipids prevent water loss and act as semiochemicals. Because of their ecological function, the profile change across the insects' sex and development offers insight into insect biology and possible tools for pest management. Here, the first work on cecidomyiid cuticular extracts is proposed considering Dasineura oleae (Diptera: Cecidomyiidae) males and females at different adult ages (0-12 h, 12-24 h, 24-36 h) and distinct sexual conditions (virgin and mated). A set of 49 compounds were recorded (12 alkanes, 1 monomethyl alkane, 11 fatty acids, 4 esters, 1 aldehyde, 1 allylbenzene, 1 amine, 1 flavonoid, 1 ketone, 1 phenol, 1 steradiene, 1 sterol, 1 terpene, 1 triterpene and 11 unknown compounds), and 18 of them showed significant differences between groups. Among alkanes, hexacosane (nC26) exhibited a decreasing trend from the youngest to the oldest females, while pentacosane (nC25) and nonacosane (nC29) showed a decreasing trend from 0 to 12 h to 12-24 h virgin females. In addition, nonadecane (nC19) was significantly more abundant in the youngest males compared to older males and females. The alkanes nC25, nC26 and nC29 have been reported to be age-related also in other dipterans, while nC19 has been described as gender-specific chemical cue for platygastrid parasitoids. Further behavioural trials and analyses are required to assign the specific ecological roles to the characterized compounds. Our results may contribute to develop new low-impact control strategies relying on the manipulation of D. oleae's chemical communication (e.g. disruption of mating or species recognition). HIGHLIGHTS: • Cuticular hydrocarbons are often involved in dipteran intraspecific communication. • We explored the cuticular profile of D. oleae at different age, sex, mating condition. • Five alkanes and one mono-methyl alkane showed differences among groups. • Linoleic acid is the most abundant compound in virgins, absent in mated insects. • Eleven compounds disappear in mated insects, but were present in all virgins.

Infestation of the gall midge Dasineura oleae provides first evidence of induced plant volatiles in olive leaves.

In this study, we present the first characterization of herbivore-induced plant volatiles (HIPVs) released from infested olive leaves. The gall midge Dasineura oleae is a specific pest of Olea europaea and endemic of the Mediterranean Basin, an area in which severe outbreaks currently occurred. Little is known about the damage caused by the pest and the relationship with its host. Since gall formation and larval feeding activity may lead to the release of specific plant volatile compounds, we investigated the volatile profiles emitted from infested plants compared with healthy plants under both laboratory and field conditions. Additionally, the volatiles emitted from mechanically damaged plants were considered. A blend of 12 volatiles was emitted from olive trees infested by D. oleae. Of these, ß-copaene, ß-ocimene, cosmene, unknown 1 and unknown 3 were found to be exclusively emitted in infested plants. The emission of germacrene-D, (E,E)-α-farnesene, and (Z,E)-α-farnesene, α-copaene, (E)-4,8-dimethylnona-1,3,7-triene, (E)-ß-guaiene and heptadecane significantly increased in infested trees. Linalool, ß-copaen-4-α-ol, ß-bourbonene, ß-cubebene, ß-elemene, ß-copaene and δ-amorphene were found only in the field trial and showed differences depending on the level of infestation and the plant stage. (Z)-3-Hexenol, (E)-4-oxohen-2-enal, and 2-(2-butoxyethoxy)-ethanol, were exclusively emitted from the leaves after mechanical damage. In a field trial in Italy, we also demonstrated spring synchronization between adults of D. oleae and O. europaea trees. Analyses of morphoanatomical malformations of gall leaves showed that tissue alterations occur at the spongy parenchyma causing an increase of the leaf blade thickness. We speculate that tissue alterations may lead to HIPV release, in turn potentially attracting D. oleae natural enemies.

Towards understanding temporal and spatial dynamics of Bactrocera oleae (Rossi) infestations using decade-long agrometeorological time series.

Insect dynamics depend on temperature patterns, and therefore, global warming may lead to increasing frequencies and intensities of insect outbreaks. The aim of this work was to analyze the dynamics of the olive fruit fly, Bactrocera oleae (Rossi), in Tuscany (Italy). We profited from long-term records of insect infestation and weather data available from the regional database and agrometeorological network. We tested whether the analysis of 13 years of monitoring campaigns can be used as basis for prediction models of B. oleae infestation. We related the percentage of infestation observed in the first part of the host-pest interaction and throughout the whole year to agrometeorological indices formulated for different time periods. A two-step approach was adopted to inspect the effect of weather on infestation: generalized linear model with a binomial error distribution and principal component regression to reduce the number of the agrometeorological factors and remove their collinearity. We found a consistent relationship between the degree of infestation and the temperature-based indices calculated for the previous period. The relationship was stronger with the minimum temperature of winter season. Higher infestation was observed in years following warmer winters. The temperature of the previous winter and spring explained 66 % of variance of early-season infestation. The temperature of previous winter and spring, and current summer, explained 72 % of variance of total annual infestation. These results highlight the importance of multiannual monitoring activity to fully understand the dynamics of B. oleae populations at a regional scale.

Evaluation of the effect of agroclimatic variables on the probability and timing of olive fruit fly attack.

Agroclimatic variables may affect insect and plant phenology, with unpredictable effects on pest populations and crop losses. Bactrocera oleae Rossi (Diptera: Tephritidae) is a specific pest of Olea europaea plants that can cause annual economic losses of more than one billion US dollars in the Mediterranean region. In this study, we aimed at understanding the effect of olive tree phenology and other agroclimatic variables on B. oleae infestation dynamics in the Umbria region (Central Italy). Analyses were carried out on B. oleae infestation data collected in 79 olive groves during a 7-year period (from 2015 to 2021). In July-August, B. oleae infestation (1% attack) was negatively affected by altitude and spring mean daily temperatures and positively by higher winter mean daily temperatures and olive tree cumulative degree days. In September-October, infestation was negatively affected by a positive soil water balance and high spring temperatures. High altitude and cumulative plant degree days were related to delayed attacks. In contrast, high winter and spring temperatures accelerated them. Our results could be helpful for the development of predictive models and for increasing the reliability of decision support systems currently used in olive orchards.

A new PCR based molecular method for early and precise quantification of parasitization in the emerging olive pest Dasineura oleae.

BACKGROUND: Dasineura oleae (Angelini 1831) (Diptera: Cecidomyiidae) was considered a minor pest in olive orchards, but in recent years severe outbreaks have been registered in several Mediterranean countries. Damage is caused by the feeding activity of larvae that induce gall formations and alters the physiological activity of the leaves. In Italy, this pest may be controlled by four Hymenoptera parasitoid species belonging to Platygaster and Mesopolobus genera such as Platygaster demades Walker 1835, Platygaster oleae Szelenyi 1940 (Hymenoptera: Platygastridae), Mesopolobus aspilus (Walker 1835) and Mesopolobus mediterraneus (Mayr 1903) (Hymenoptera: Pteromalidae), but parasitization becomes evident only after gall dissection. RESULTS: In this study, we aim to: (i) design a primer for the detection of specimens belonging to Platygaster and Mesopolobus genera; (ii) develop a multiplex quantitative polymerase chain reaction (qPCR) protocol combined to a fast samples DNA extraction method; (iii) apply the developed protocol to field-collected specimens and compare this method with traditional techniques based on visual estimation of parasitism rate on larvae. Primers were designed to anneal with cytochrome oxidase subunit I (COI) sequences of Platygaster and Mesopolobus genera while protocols were developed to be fast and capable to process several samples at the same time. Molecular analyses demonstrated to provide almost double of the parasitism rate assessed by visual inspection. Furthermore, on second instar larvae the PCR-based method was able to detect ten-fold times the parasitization rate estimated by visual inspection. CONCLUSION: The application on a greater scale of this newly developed method could be fundamental in the determination of the biological control potential in olive orchards.

Climate Change and Major Pests of Mediterranean Olive Orchards: Are We Ready to Face the Global Heating?

Evidence of the impact of climate change on natural and agroecosystems is nowadays established worldwide, especially in the Mediterranean Basin, an area known to be very susceptible to heatwaves and drought. Olea europaea is one of the main income sources for the Mediterranean agroeconomy, and it is considered a sensitive indicator of the climate change degree because of the tight relationship between its biology and temperature trend. Despite the economic importance of the olive, few studies are nowadays available concerning the consequences that global heating may have on its major pests. Among the climatic parameters, temperature is the key one influencing the relation between the olive tree and its most threatening parasites, including Bactrocera oleae and Prays oleae. Therefore, several prediction models are based on this climatic parameter (e.g., cumulative degree day models). Even if the use of models could be a promising tool to improve pest control strategies and to safeguard the Mediterranean olive patrimony, they are not currently available for most O. europaea pests, and they have to be used considering their limits. This work stresses the lack of knowledge about the biology and the ethology of olive pests under a climate change scenario, inviting the scientific community to focus on the topic.

Ultrastructural characterization of antennal sensilla and immunocytochemical localization of a chemosensory protein in Carausius morosus Brünner (Phasmida: Phasmatidae).

The aim of this work was to investigate the olfactory system of the walking stick insect, Carausius morosus. Morphological, ultrastructural and immunocytochemical studies of adult female antennae were conducted by scanning and transmission electron microscopy. Extensive cross-section series were made through the last antennal segment to define the cuticular apparatus, wall pore distribution and the number of innervating receptor neurons of each sensillum type. Single-walled wall pore sensilla occur in three subtypes: (i) with 27 or 28 branched receptor neurons, (ii) with two branched neurons and (iii) with one or two unbranched neurons, respectively. Double-walled wall pore sensilla were found in two subtypes with spoke channels, one with four unbranched neurons, the other with two unbranched neurons. One terminal pore sensillum was found, showing two cavities within the hair and being innervated by six sensory cells. Immunocytochemical experiments were performed to show the localization of a 19 kDa soluble protein found in the chemosensory organs of C. morosus. This protein shows an amino acid sequence homologous to the family of chemosensory proteins (CSP). The polyclonal antibody raised against the purified protein (CSP-cmA) showed, for the first time in CSPs, a strong labeling in olfactory sensilla, specifically in the sensillum lymph surrounding the dendritic branches of SW-WP sensilla and in the uninnervated lumen between the two concentric walls of DW-WP type 1 sensilla.