Effective Stochastic Algorithm in Disease Prediction.

Traditionally, the main process for olive fruit fly population monitoring is trap measurements. Although the above procedure is time-consuming, it gives important information about when there is an outbreak of the population and how the insect is spatially distributed in the olive grove. Most studies in the literature are based on the combination of trap and environmental data measurements. Strictly speaking, the dynamics of olive fruit fly population is a complex system affected by a variety of factors. However, the collection of environmental data is costly, and sensor data often require additional processing and cleaning. In order to study the volatility of correlation in trap counts and how it is connected with population outbreaks, a stochastic algorithm, based on a stochastic differential model, is experimentally applied. The results allow us to predict early population outbreaks allowing for more efficient and targeted spraying.

The Effect of Olive Fruit Fly Bactrocera oleae (Rossi) Infestation on Certain Chemical Parameters of Produced Olive Oils.

Olives affected by active and damaging infestation (olive fruit fly Bactrocera oleae (Rossi)) were assayed for their chemical composition. Biophenols were determined by HPLC, sterols, triterpenic dialcohols, and fatty acids by gas chromatography analysis. The acquired data were statistically analyzed. Oils produced from "Istrska belica" fruit affected by active infestation compared to the oils made from fruit affected by damaging infestation showed higher amounts of total oleuropein biofenols (377.3 versus (vs.) 106.6 mg/kg), total biophenols (755 vs. 377 mg/kg), lignans (85.3 vs. 32.9 mg/kg), the dialdehydic form of decarboxymethyl oleuropein aglycone (DMO-Agl-dA) (148.3 vs. 49.0 mg/kg), its oxidized form (DMO-Agl-dA)ox (35.2 vs. 8.5 mg/kg), the dialdehydic form of oleuropein aglycone (O-Agl-dA) (61.1 vs. 8.0 mg/kg), the dialdehydic form of ligstroside aglycone (L-Agl-dA) (63.5 vs. 28.0 mg/kg), the aldehydic form of oleuropein aglycone (O-Agl-A) (40.6 vs. 8.4 mg/kg), and lower amounts of tyrosol (Tyr) (6.0 vs. 13. 9 mg/kg) and the aldehydic form of ligstroside aglycone (L-Agl-A) (13.8 vs. 40.3 mg/kg). Higher values of stigmasterol (2.99%) and lower values of campesterol (2.25%) were determined in oils affected by damaging infestation; an increase in triterpenic dialcohols was also observed (3.04% for damaging and 1.62% for active infestation). Oils affected by damaging infestation, compared to active infestation, showed lower amounts of oleic acid (73.89 vs. 75.15%) and higher amounts of myristic (0.013 vs. 0.011%), linoleic (7.27 vs. 6.48%), and linolenic (0.74 vs. 0.61%) acids.

Olive fruit fly rearing procedures affect the vertical transmission of the bacterial symbiont Candidatus Erwinia dacicola.

BACKGROUND: The symbiosis between the olive fruit fly, Bactrocera oleae, and Candidatus Erwinia dacicola has been demonstrated as essential for the fly's larval development and adult physiology. The mass rearing of the olive fruit fly has been hindered by several issues, including problems which could be related to the lack of the symbiont, presumably due to preservatives and antibiotics currently used during rearing under laboratory conditions. To better understand the mechanisms underlying symbiont removal or loss during the rearing of lab colonies of the olive fruit fly, we performed experiments that focused on bacterial transfer from wild female flies to their eggs. In this research, eggs laid by wild females were treated with propionic acid solution, which is often used as an antifungal agent, a mixture of sodium hypochlorite and Triton X, or water (as a control). The presence of the bacterial symbiont on eggs was evaluated by real-time PCR and scanning electron microscopy. RESULTS: DGGE analysis showed a clear band with the same migration behavior present in all DGGE profiles but with a decreasing intensity. Molecular analyses performed by real-time PCR showed a significant reduction in Ca. E. dacicola abundance in eggs treated with propionic acid solution or a mixture of sodium hypochlorite and Triton X compared to those treated with water. In addition, the removal of bacteria from the surfaces of treated eggs was highlighted by scanning electron microscopy. CONCLUSIONS: The results clearly indicate how the first phases of the colony-establishment process are important in maintaining the symbiont load in laboratory populations and suggest that the use of products with antimicrobial activity should be avoided. The results also suggest that alternative rearing procedures for the olive fruit fly should be investigated.

Horizontal transfer and finalization of a reliable detection method for the olive fruit fly endosymbiont, Candidatus Erwinia dacicola.

BACKGROUND: The olive fly, Bactrocera oleae, is the most important insect pest in olive production, causing economic damage to olive crops worldwide. In addition to extensive research on B. oleae control methods, scientists have devoted much effort in the last century to understanding olive fly endosymbiosis with a bacterium eventually identified as Candidatus Erwinia dacicola. This bacterium plays a relevant role in olive fly fitness. It is vertically transmitted, and it benefits both larvae and adults in wild populations; however, the endosymbiont is not present in lab colonies, probably due to the antibiotics and preservatives required for the preparation of artificial diets. Endosymbiont transfer from wild B. oleae populations to laboratory-reared ones allows olive fly mass-rearing, thus producing more competitive flies for future Sterile Insect Technique (SIT) applications. RESULTS: We tested the hypothesis that Ca. E. dacicola might be transmitted from wild, naturally symbiotic adults to laboratory-reared flies. Several trials have been performed with different contamination sources of Ca. E. dacicola, such as ripe olives and gelled water contaminated by wild flies, wax domes containing eggs laid by wild females, cages dirtied by faeces dropped by wild flies and matings between lab and wild adults. PCR-DGGE, performed with the primer set 63F-GC/518R, demonstrated that the transfer of the endosymbiont from wild flies to lab-reared ones occurred only in the case of cohabitation. CONCLUSIONS: Cohabitation of symbiotic wild flies and non-symbiotic lab flies allows the transfer of Ca. E. dacicola through adults. Moreover, PCR-DGGE performed with the primer set 63F-GC/518R was shown to be a consistent method for screening Ca. E. dacicola, also showing the potential to distinguish between the two haplotypes (htA and htB). This study represents the first successful attempt at horizontal transfer of Ca. E. dacicola and the first step in acquiring a better understanding of the endosymbiont physiology and its relationship with the olive fly. Our research also represents a starting point for the development of a laboratory symbiotic olive fly colony, improving perspectives for future applications of the Sterile Insect Technique.

Barcoding of parasitoid wasps (Braconidae and Chalcidoidea) associated with wild and cultivated olives in the Western Cape of South Africa 1.

Wild and cultivated olives harbor and share a diversity of insects, some of which are considered agricultural pests, such as the olive fruit fly. The assemblage of olive-associated parasitoids and seed wasps is rich and specialized in sub-Saharan Africa, with native species possibly coevolving with their hosts. Although historical entomological surveys reported on the diversity of olive wasp species in the Western Cape Province of South Africa, no comprehensive study has been performed in the region in the molecular era. In this study, a dual approach combining morphological and DNA-based methods was used for the identification of adult specimens reared from olive fruits. Four species of Braconidae and six species of Chalcidoidea were identified, and DNA barcoding methodologies were used to investigate conspecificity among individuals, based on randomly selected representative specimens. Morphological identifications were congruent with DNA data, as NJ and ML trees correctly placed the sequences for each species either at the genus or species level, depending on the available taxa coverage, and genetic distances strongly supported conspecificity. No clear evidence of cryptic diversity was found. Overall seed infestation and parasitism rates were higher in wild olives compared to cultivated olives, and highest for Eupelmus spermophilus and Utetes africanus. These results can be used for early DNA-based detection of wasp larvae in olives and to further investigate the biology and ecology of these species.

Evaluation of the Phytopathological Reaction of Wild and Cultivated Olives as a Means of Finding Promising New Sources of Genetic Diversity for Resistance to Root-Knot Nematodes.

Olive (Olea europaea L.) is one of the most important fruit crops in the Mediterranean Basin, because it occupies significant acreage in these countries and often has important cultural heritage and landscape value. This crop can be infected by several Meloidogyne species (M. javanica, M. arenaria, and M. incognita, among others), and only a few cultivars with some level of resistance to these nematodes have been found. Innovations in intensive olive growing using high planting densities, irrigation, and substantial amounts of fertilizers could increase the nematode population to further damaging levels. To further understand the interactions involved between olive and pathogenic nematodes and in the hope of finding solutions to the agricultural risks, this research aimed to determine the reaction of important olive cultivars in Spain and wild olives to M. javanica infection, including genotypes of the same and other O. europaea subspecies. All olive cultivars tested were good hosts for M. javanica, but high levels of nematode reproduction found in three cultivars (Gordal Sevillana, Hojiblanca, and Manzanilla de Sevilla) were substantially different. In the wild accessions, O. europaea subsp. cerasiformis (genotype W147) and O. europaea subsp. europaea var. sylvestris (genotype W224) were resistant to M. javanica at different levels, with strong resistance in W147 (reproduction factor [Rf] = 0.0003) and moderate resistance in W224 (Rf = 0.79). The defense reaction of W147 to M. javanica showed a strong increase of phenolic compounds but no hypersensitive reaction.

Bio-insecticidal effects of Oleaster leaves aqueous extracts against Psylla larvae (Euphyllura olivina (Costa)), a primary pest of Olea europaea L.

Many plant species produce phenolic compounds in their various organs and their use in crop protection. These plant secondary metabolites may serve as toxins against the insect pests. The objective of this study was to evaluate in vitro the bio-insecticidal effect of an aqueous extract of wild Olive leaves on Psylla larvae (Euphyllura olivina), a primary pest of the cultivated Olive tree (Olea europaea L. subsp sativa). Two concentrations of 0.05g/ml and 0.1g/ml leaves grinding powder in distilled water were sprayed on branches infested with Psylla larvae. The obtained results revealed a very significant mortality rate of the larvae 24 hours after spraying. The chemical composition of Oleaster leaves aqueous extracts is determined by HPLC-DAD. The results show in majority the presence of phenolic compounds represented by oleuropein and its metabolite hydroxytyrosol. The phenolic compounds of the crude extract were at the origin of this mortality. The Analysis of Variance revealed highly significant results both between the sampled trees and between the tested concentrations. The Principal Component Analysis (PCA) revealed a close relation between the physiological state of the studied trees and the degree of their infestation by the phytophagus. Taking into account, the physical and chemical characteristics of the sampled soils, data analysis showed that trees growing on nitrogen-rich soils were more infested than those growing on soils rich in organic carbon (Corg) and phosphorus (Porg).

Diversity of scale insects (Hemiptera, Coccomorpha) in ten varieties of olive trees (Olea europaea L.) in Southern Brazil.

Olive production (Olea europaea L.) is a recent activity in Brazil and studies on the adaptability of olive trees varieties in the States are required. One of the problems that can hamper the development of olive trees is the incidence of pests, such as scale insects (Hemiptera, Coccomorpha). In this study, the associated scale insect species were found in ten varieties in an olive grove located at the Research Center of the Serra do Sudeste Region (30º33'15"S, 52º23'45"W) in Brazil. The surveys were carried out twice in each season (2013-2014). Six species of armored scale insect (Diaspididae) and two soft scales (Coccidae) were determined. Saissetia oleae (Coccidae) with 88.15% and Aspidiotus nerii (Diaspididae) with 5.85% occurred in all varieties of olive trees. The varieties Arbequina, Frantoio and Cipressino were with greater richness of scale insects, the highest values of diversity occurred in Frantoio, Arbequina and Picual; in Alfafara, Arbosana and Coratina there was a high dominance of one species (S. oleae).

Molecular characterization of pyrethroid resistance in the olive fruit fly Bactrocera oleae.

Α reduction of pyrethroid efficacy has been recently recorded in Bactrocera oleae, the most destructive insect of olives. The resistance levels of field populations collected from Crete-Greece scaled up to 22-folds, compared to reference laboratory strains. Sequence analysis of the IIS4-IIS6 region of para sodium channel gene in a large number of resistant flies indicated that resistance may not be associated with target site mutations, in line with previous studies in other Tephritidae species. We analyzed the transcriptomic differences between two resistant populations versus an almost susceptible field population and two laboratory strains. A large number of genes was found to be significantly differentially transcribed across the pairwise comparisons. Interestingly, gene set analysis revealed that genes of the 'electron carrier activity' GO group were enriched in one specific comparison, which might suggest a P450-mediated resistance mechanism. The up-regulation of several transcripts encoding detoxification enzymes was qPCR validated, focusing on transcripts coding for P450s. Of note, the expression of contig00436 and contig02103, encoding CYP6 P450s, was significantly higher in all resistant populations, compared to susceptible ones. These results suggest that an increase in the amount of the CYP6 P450s might be an important mechanism of pyrethroid resistance in B. oleae.

How anthropogenic changes may affect soil-borne parasite diversity? Plant-parasitic nematode communities associated with olive trees in Morocco as a case study.

BACKGROUND: Plant-parasitic nematodes (PPN) are major crop pests. On olive (Olea europaea), they significantly contribute to economic losses in the top-ten olive producing countries in the world especially in nurseries and under cropping intensification. The diversity and the structure of PPN communities respond to environmental and anthropogenic forces. The olive tree is a good host plant model to understand the impact of such forces on PPN diversity since it grows according to different modalities (wild, feral and cultivated olives). A wide soil survey was conducted in several olive-growing regions in Morocco. The taxonomical and the functional diversity as well as the structures of PPN communities were described and then compared between non-cultivated (wild and feral forms) and cultivated (traditional and high-density olive cultivation) olives. RESULTS: A high diversity of PPN with the detection of 117 species and 47 genera was revealed. Some taxa were recorded for the first time on olive trees worldwide and new species were also identified. Anthropogenic factors (wild vs cultivated conditions) strongly impacted the PPN diversity and the functional composition of communities because the species richness, the local diversity and the evenness of communities significantly decreased and the abundance of nematodes significantly increased in high-density conditions. Furthermore, these conditions exhibited many more obligate and colonizer PPN and less persister PPN compared to non-cultivated conditions. Taxonomical structures of communities were also impacted: genera such as Xiphinema spp. and Heterodera spp. were dominant in wild olive, whereas harmful taxa such as Meloidogyne spp. were especially enhanced in high-density orchards. CONCLUSIONS: Olive anthropogenic practices reduce the PPN diversity in communities and lead to changes of the community structures with the development of some damaging nematodes. The study underlined the PPN diversity as a relevant indicator to assess community pathogenicity. That could be taken into account in order to design control strategies based on community rearrangements and interactions between species instead of reducing the most pathogenic species.

Diversity of root-knot nematodes in Moroccan olive nurseries and orchards: does Meloidogyne javanica disperse according to invasion processes?

BACKGROUND: Root-knot nematodes (RKN) are major pest of olive tree (Olea europaea ssp. europaea), especially in nurseries and high-density orchards. Soil samples were collected from main olive growing areas of Morocco, to characterize Meloidogyne species and to discuss the contribution of biotic and abiotic factors in their spatial distribution. RESULTS: RKN were found in 159 soil samples out of 305 from nurseries (52.1% occurrence) and in 11 out of 49 soil samples from orchards (23.2% occurrence). Biochemical and molecular characterisation (PAGE esterase and SCAR) revealed the dominance of M. javanica both in nurseries and orchards with minor presence of M. incognita only in nurseries, and M. arenaria in only one nursery. RKN were distributed on aggregated basis. Frequent presence of M. javanica in orchards might have come from nurseries. In contrast, the detection of M. incognita in nurseries alone suggests that this species could not reproduce in orchards because of either the competition with other plant-parasitic nematodes or unfit local habitats. The impact of environmental variables (climate, habitat origin and physicochemical characteristics of the substrates) on the distribution of Meloidogyne species is also discussed. CONCLUSION: Olive nurseries in Morocco are not able to guarantee the safety of rooted plants. As a result, olive production systems are exposed to strong RKN invasion risks. Consequently, the use of healthy substrates in nurseries may prevent plant-parasitic nematode induction in orchards.

Fine-scale ecological and economic assessment of climate change on olive in the Mediterranean Basin reveals winners and losers.

The Mediterranean Basin is a climate and biodiversity hot spot, and climate change threatens agro-ecosystems such as olive, an ancient drought-tolerant crop of considerable ecological and socioeconomic importance. Climate change will impact the interactions of olive and the obligate olive fruit fly (Bactrocera oleae), and alter the economics of olive culture across the Basin. We estimate the effects of climate change on the dynamics and interaction of olive and the fly using physiologically based demographic models in a geographic information system context as driven by daily climate change scenario weather. A regional climate model that includes fine-scale representation of the effects of topography and the influence of the Mediterranean Sea on regional climate was used to scale the global climate data. The system model for olive/olive fly was used as the production function in our economic analysis, replacing the commonly used production-damage control function. Climate warming will affect olive yield and fly infestation levels across the Basin, resulting in economic winners and losers at the local and regional scales. At the local scale, profitability of small olive farms in many marginal areas of Europe and elsewhere in the Basin will decrease, leading to increased abandonment. These marginal farms are critical to conserving soil, maintaining biodiversity, and reducing fire risk in these areas. Our fine-scale bioeconomic approach provides a realistic prototype for assessing climate change impacts in other Mediterranean agro-ecosystems facing extant and new invasive pests.

Evaluation of native plant flower characteristics for conservation biological control of Prays oleae.

Several studies have shown that manipulating flowering weeds within an agroecosystem can have an important role in pest control by natural enemies, by providing them nectar and pollen, which are significant sources of nutrition for adults. The aim of this study was to assess if the olive moth, Prays oleae (Bernard, 1788) (Lepidoptera: Praydidae), and five of its main natural enemies, the parasitoid species Chelonus elaeaphilus Silvestri (Hymenoptera: Braconidae), Apanteles xanthostigma (Haliday) (Hymenoptera: Braconidae), Ageniaspis fuscicollis (Dalman) (Hymenoptera: Encyrtidae) and Elasmus flabellatus (Fonscolombe) (Hymenoptera: Eulophidae), as well as the predator Chrysoperla carnea (Stephens) (Neuroptera: Chrysopidae), can theoretically access the nectar from 21 flowering weeds that naturally occur in olive groves. Thus, the architecture of the flowers as well as the mouthpart structure and/or the head and thorax width of the pest and its enemies were analyzed. The results suggested that all beneficial insects were able to reach nectar of the plant species from Apiaceae family, i.e. Conopodium majus (Gouan) Loret, Daucus carota L. and Foeniculum vulgare Mill., as well as Asparagus acutifolius L., Echium plantagineum L., Capsella bursa-pastoris (L.) Medik., Raphanus raphanistrum L., Lonicera hispanica Boiss. et Reut., Silene gallica L., Spergula arvensis L., Hypericum perforatum L., Calamintha baetica Boiss. et Reut, Malva neglecta Wallr. and Linaria saxatilis (L.) Chaz. P. oleae was not able to access nectar from five plant species, namely: Andryala integrifolia L., Chondrilla juncea L., Dittrichia viscosa (L.) Greuter, Sonchus asper (L.) Hill and Lavandula stoechas L.

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.

Olive fruits infested with olive fly larvae respond with an ethylene burst and the emission of specific volatiles.

Olive fly (Bactrocera oleae R.) is the most harmful insect pest of olive (Olea europaea L.) which strongly affects fruits and oil production. Despite the expanding economic importance of olive cultivation, up to now, only limited information on plant responses to B. oleae is available. Here, we demonstrate that olive fruits respond to B. oleae attack by producing changes in an array of different defensive compounds including phytohormones, volatile organic compounds (VOCs), and defense proteins. Bactrocera oleae-infested fruits induced a strong ethylene burst and transcript levels of several putative ethylene-responsive transcription factors became significantly upregulated. Moreover, infested fruits induced significant changes in the levels of 12-oxo-phytodienoic acid and C12 derivatives of the hydroperoxide lyase. The emission of VOCs was also changed quantitatively and qualitatively in insect-damaged fruits, indicating that B. oleae larval feeding can specifically affect the volatile blend of fruits. Finally, we show that larval infestation maintained high levels of trypsin protease inhibitors in ripe fruits, probably by affecting post-transcriptional mechanisms. Our results provide novel and important information to understand the response of the olive fruit to B. oleae attack; information that can shed light onto potential new strategies to combat this pest.

Aldehyde-containing clays: a sustainable approach against the olive tree pest, Bactrocera oleae.

A set of organic/inorganic layered materials was obtained by functionalizing a montmorillonite-containing bentonite natural clay with linear aliphatic C6 or C7 aldehydes through a cost-effective and technologically simple incipient-wetness deposition method. The solids were investigated by means of a multi-technique approach (X-ray powder diffraction, XRPD, scanning electron microscopy, SEM, Fourier-transform infrared spectroscopy, FT-IR, thermogravimetric analysis, TGA, elemental analysis and solid-state nuclear magnetic resonance, ssNMR) to clarify the nature of the deposited organic species and the mode of interaction between the aldehyde and the clay. Since both natural clays and short-chain linear aldehydes find application as alternative strategies in the control of the olive fruit fly, Bactrocera oleae, the hybrid layered materials were tested under real-life conditions and their insect-inhibiting capability was evaluated in open-field trials on olive tree orchards in Tuscany, Central Italy. Specific tests were conducted to evaluate the resistance of the solids to weathering and their capability to provide a constant and long-lasting release of the bioactive ingredient. Aldehyde-containing bentonite clays have shown promising performance in controlling B. oleae infestation (with up to 86-95% reduction of affected olive fruits) in open-field trials across two years in two locations with different pedological and meteo-climatic characteristics.

Control of the olive fruit fly using genetics-enhanced sterile insect technique.

BACKGROUND: The olive fruit fly, Bactrocera oleae, is the major arthropod pest of commercial olive production, causing extensive damage to olive crops worldwide. Current control techniques rely on spraying of chemical insecticides. The sterile insect technique (SIT) presents an alternative, environmentally friendly and species-specific method of population control. Although SIT has been very successful against other tephritid pests, previous SIT trials on olive fly have produced disappointing results. Key problems included altered diurnal mating rhythms of the laboratory-reared insects, resulting in asynchronous mating activity between the wild and released sterile populations, and low competitiveness of the radiation-sterilised mass-reared flies. Consequently, the production of competitive, male-only release cohorts is considered an essential prerequisite for successful olive fly SIT. RESULTS: We developed a set of conditional female-lethal strains of olive fly (named Release of Insects carrying a Dominant Lethal; RIDL®), providing highly penetrant female-specific lethality, dominant fluorescent marking, and genetic sterility. We found that males of the lead strain, OX3097D-Bol, 1) are strongly sexually competitive with wild olive flies, 2) display synchronous mating activity with wild females, and 3) induce appropriate refractoriness to wild female re-mating. Furthermore, we showed, through a large proof-of-principle experiment, that weekly releases of OX3097D-Bol males into stable populations of caged wild-type olive fly could cause rapid population collapse and eventual eradication. CONCLUSIONS: The observed mating characteristics strongly suggest that an approach based on the release of OX3097D-Bol males will overcome the key difficulties encountered in previous olive fly SIT attempts. Although field confirmation is required, the proof-of-principle suppression and elimination of caged wild-type olive fly populations through OX3097D-Bol male releases provides evidence for the female-specific RIDL approach as a viable method of olive fly control. We conclude that the promising characteristics of OX3097D-Bol may finally enable effective SIT-based control of the olive fly.

Morphological and molecular characterisation of Xiphinema macroacanthum Lamberti, Roca & Agostinelli, 1989 (Nematoda: Longidoridae) from olive orchards in southern Italy.

A population of Xiphinema macroacanthum Lamberti, Roca & Agostinelli, 1989 originating from olive orchards in Brindisi, Italy and containing both adults and all juvenile stages, is described and illustrated. The first juvenile stage is reported for the first time. Molecular characterisation of this species, using the D2-D3 expansion domains of the 28S rDNA and ITS region, was carried out. PCR-RFLP analyses of the ribosomal gene regions determined species-specific patterns that clearly differentiate X. macroacanthum. Sequences of the D2-D3 domains and the partial 18S-ITS1 rRNA genes were analysed using several methods for inferring phylogeny to reconstruct the relationships between X. macroacanthum and other Xiphinema species.

Some ecological aspects on olive parlatoria scale, Parlatoria oleae (Colvee) infested plum and olive trees under irrigation system at Burg El-Arab area, Alexandria, Egypt.

Field studies were conducted under irrigation system at Burg El-Arab area (50 K.m. west of Alexandria city) to study some ecological aspects of Parlatoria oleae (Colvee) and its parasitoid, Aphytis maculicornis (Masi) that infested olive and plum trees during the two successive years of 2010 and 2011. Field observations revealed that the most of the inspected individuals occurred mainly on the branches of plum trees as compared with those found on leaves, but on olive the insect occurred on all the plant parts including fruits. On plum trees, the obtained data showed that the population of P. oleae reached the maximum during April, November and January in the first year, but in the second one it had four peaks of high population levels during March, August, November and January. The maximal percent as adult females was observed during March, September and January, whereas the adult males appeared only during October and November. The parasitized individuals by A. maculicornis represented higher rate of parasitization from March to August in the first year that extended to September in the second year. On olive trees under irrigation the population of Parlatoria scale and its parasitoid was relatively low. It had three high peaks, during March, December and February in the first year, while in the second one the population reached the maximum during March, December and February. The parasitoid Aphytis maculicormis appeared in March and April marking high percentage levels in the first year, noticeably it was nearly parasitized the scale insect all the year round.

Fungal diversity associated to the olive moth, Prays Oleae bernard: a survey for potential entomopathogenic fungi.

Olive production is one of the main agricultural activities in Portugal. In the region of Trás-os-Montes, this crop has been considerably affected by Prays oleae. In order to evaluate the diversity of fungi on Prays oleae population of Trás-os-Montes olive orchards, larvae and pupae of the three annual generations (phyllophagous, antophagous and carpophagous) were collected and evaluated for fungal growth on their surface. From the 3,828 larvae and pupae, a high percentage of individuals exhibited growth of a fungal agent (40.6%), particularly those from the phyllophagous generation. From all the moth generations, a total of 43 species from 24 genera were identified, but the diversity and abundance of fungal species differed between the three generations. Higher diversity was found in the carpophagous generation, followed by the antophagous and phyllophagous generations. The presence of fungi displaying entomopathogenic features was highest in the phyllophagous larvae and pupae, with Beauveria bassiana as the most abundant taxa. The first report of Beauveria bassiana presence on Prays oleae could open new strategies for the biocontrol of this major pest in olive groves since the use of an already adapted species increases the guarantee of success of a biocontrol approach. The identification of antagonistic fungi able to control agents that cause major olive diseases, such as Verticillium dahliae, will benefit future biological control approaches for limiting this increasingly spreading pathogen.