Volatile allosteric antagonists of mosquito odorant receptors inhibit human-host attraction.

Odorant-dependent behaviors in insects are triggered by the binding of odorant ligands to the variable subunits of heteromeric olfactory receptors. Previous studies have shown, however, that specific odor binding to ORco, the common subunit of odorant receptor heteromers, may allosterically alter olfactory receptor function and profoundly affect subsequent behavioral responses. Using an insect cell-based screening platform, we identified and characterized several antagonists of the odorant receptor coreceptor of the African malaria vector Anopheles gambiae (AgamORco) in a small collection of natural volatile organic compounds. Because some of the identified antagonists were previously shown to strongly repel Anopheles and Culex mosquitoes, we examined the bioactivities of the identified antagonists against Aedes, the third major genus of the Culicidae family. The tested antagonists inhibited the function of Ae. aegypti ORco ex vivo and repelled adult Asian tiger mosquitoes (Ae. albopictus). Binary mixtures of specific antagonists elicited higher repellency than single antagonists, and binding competition assays suggested that this enhanced repellence is due to antagonist interaction with distinct ORco sites. Our results also suggest that the enhanced mosquito repellency by antagonist mixtures is due to additive rather than synergistic effects of the specific antagonist combinations on ORco function. Taken together, these findings provide novel insights concerning the molecular aspects of odorant receptor function. Moreover, our results demonstrate that a simple screening assay may be used for the identification of allosteric modifiers of olfactory-driven behaviors capable of providing enhanced personal protection against multiple mosquito-borne infectious diseases.

Sublethal effects of oregano essential oil and its major compound carvacrol on biological parameters of Aedes albopictus (Diptera: Culicidae).

Mosquito management programs rely basically on the use of conventional synthetic larvicides. However, frequent applications and misuse of some synthetic insecticides have led to problems related to mosquito resistance development, harmful effects on human health and unacceptable environmental effects on non-target organisms. Recently, a growing number of phytochemicals has been tested as more eco-friendly larvicides against various mosquito species, exerting high efficacy with multiple modes of action. In the laboratory, we investigated for the first time the sublethal effects of oregano oil and its major compound carvacrol, against Aedes albopictus (Asian tiger mosquito), a mosquito of great medical importance. We determined the effects of short term (24h) exposure of 3rd- 4th larvae to LC50 concentrations on survival and development of survived larvae until adulthood, as well as on fecundity, fertility, longevity and wing length of emerged adults. Only half of 24h survived larvae from oregano oil and carvacrol treatment finally reached adulthood. Abnormal shapes of dead larvae and pupae, and failed adult emergence were also observed, indicating a potential growth inhibitory activity of the tested materials. No particular effects from exposure to larvicidal LC50 concentrations were recorded on life cycle parameters of successfully emerged adults. These findings suggest the tested oregano oil and carvacrol as sufficiently effective larvicides against Ae. albopictus at lower than the acutely toxic concentrations, promoting a more eco-friendly and less costly profile for these biopesticides.

Curcumin Derivatives as Potential Mosquito Larvicidal Agents against Two Mosquito Vectors, Culex pipiens and Aedes albopictus.

Vector-borne diseases have appeared or re-emerged in many Southern Europe countries making the transmission of infectious diseases by mosquitoes (vectors) one of the greatest worldwide health threats. Larvicides have been used extensively for the control of Aedes (Stegomyia) albopictus (Skuse, 1895) (Diptera: Culicidae) and Culex pipiens Linnaeus, 1758 (Diptera: Culicidae) mosquitoes in urban and semi-urban environments, causing the increasing resistance of mosquitoes to commercial insecticides. In this study, 27 curcuminoids and monocarbonyl curcumin derivatives were synthesised and evaluated as potential larvicidal agents against Cx. pipiens and Ae. albopictus. Most of the compounds were more effective against larvae of both mosquito species. Four of the tested compounds, curcumin, demethoxycurcumin, curcumin-BF2 complex and a monocarbonyl tetramethoxy curcumin derivative exhibited high activity against both species. In Cx. pipiens the recorded LC50 values were 6.0, 9.4, 5.0 and 32.5 ppm, respectively, whereas in Ae. albopictus they exhibited LC50 values of 9.2, 36.0, 5.5 and 23.6 ppm, respectively. No conclusive structure activity relationship was evident from the results and the variety of descriptors values generated in silico provided some insight to this end.

Isolation of Volatile Compounds with Repellent Properties against Aedes albopictus (Diptera: Culicidae) Using CPC Technology.

The present work describes the use of Centrifugal Partition Chromatography (CPC) for the bio-guided isolation of repellent active volatile compounds from essential oils. Five essential oils (EOs) obtained from three Pinus and two Juniperus species were initially analyzed by gas chromatography-mass spectrometry (GC/MS) and evaluated for their repellent properties against Aedes albopictus. The essential oil from needles of P. pinea (PPI) presented the higher activity, showing 82.4% repellency at a dose of 0.2 µL/cm2. The above EO, together with the EO from the fruits of J. oxycedrus subsp. deltoides (JOX), were further analyzed by CPC using the biphasic system n-Heptane/ACN/BuOH in ratio 1.6/1.6/0.2 (v/v/v). The analysis of PPI essential oil resulted in the recovery of (-)-limonene, guaiol and simple mixtures of (-)-limonene/ß-pheladrene, while the fractionation of JOX EO led to the recovery of ß-myrcene, germacrene-D, and mixtures of α-pinene/ß-pinene (ratio 70/30) and α-pinene/germacrene D (ratio 65/45). All isolated compounds and recovered mixtures were tested for their repellent activity. From them, (-)-limonene, guaiol, germacrene-D as well the mixtures of (-)-limonene/ß-pheladrene presented significant repellent activity (>97% repellency) against Ae. albopictus. The present methodology could be a valuable tool in the effort to develop potent mosquito repellents which are environmentally friendly.

An essay on ecosystem availability of Nicotiana glauca graham alkaloids: the honeybees case study.

BACKGROUND: Invasive plant species pose a significant threat for fragile isolated ecosystems, occupying space, and consuming scarce local resources. Recently though, an additional adverse effect was recognized in the form of its secondary metabolites entering the food chain. The present study is elaborating on this subject with a specific focus on the Nicotiana glauca Graham (Solanaceae) alkaloids and their occurrence and food chain penetrability in Mediterranean ecosystems. For this purpose, a targeted liquid chromatography electrospray tandem mass spectrometric (LC-ESI-MS/MS) analytical method, encompassing six alkaloids and one coumarin derivative, utilizing hydrophilic interaction chromatography (HILIC) was developed and validated. RESULTS: The method exhibited satisfactory recoveries, for all analytes, ranging from 75 to 93%, and acceptable repeatability and reproducibility. Four compounds (anabasine, anatabine, nornicotine, and scopoletin) were identified and quantified in 3 N. glauca flowers extracts, establishing them as potential sources of alien bio-molecules. The most abundant constituent was anabasine, determined at 3900 µg/g in the methanolic extract. These extracts were utilized as feeding treatments on Apis mellifera honeybees, resulting in mild toxicity documented by 16-18% mortality. A slightly increased effect was elicited by the methanolic extract containing anabasine at 20 µg/mL, where mortality approached 25%. Dead bees were screened for residues of the N. glauca flower extracts compounds and a significant mean concentration of anabasine was evidenced in both 10 and 20 µg/mL treatments, ranging from 51 to 92 ng/g per bee body weight. Scopoletin was also detected in trace amounts. CONCLUSIONS: The mild toxicity of the extracts in conjunction with the alkaloid and coumarin residual detection in bees, suggest that these alien bio-molecules are transferred within the food chain, suggesting a chemical invasion phenomenon, never reported before.

Chemical composition and assessment of larvicidal and repellent capacity of 14 Lamiaceae essential oils against Aedes albopictus.

In the current laboratory study, 14 essential oils (EOs) derived from 12 Lamiaceae plant species and their major components were screened for their larvicidal and repellent properties against Aedes albopictus, an invasive mosquito species of great medical importance. The results of toxicity bioassays revealed that the EOs from Thymus vulgaris, Ocimum basilicum, Origanum dictamnus, Origanum majorana, and Origanum vulgare, as well as their major components (terpenes), namely thymol, carvacrol, p-cymene, and γ-terpinene exerted the highest larvicidal effect. Essential oils from Mellisa officinalis, Origanum dictamus, Mentha spicata (chem. piperitenone epoxide), Origanum majorana, and Satureja thymbra were the most potent repellents, with the last two assigned as the best ones. Among the terpenes tested, piperitenone epoxide, carvacrol, thymol, and piperitenone provided the highest level of protection against Ae. albopictus adults. Chemical analysis revealed the presence of a high number of terpenes in the EOs, while in most cases, the biological action of the tested EOs and their major components was in consistency. The most effective EOs and terpenes that were identified through the current laboratory bioassays could be used as alternative agents to control larvae and repel adults of Ae. albopictus.

Greek Pinus essential oils: larvicidal activity and repellency against Aedes albopictus (Diptera: Culicidae).

The needle volatiles metabolites of seven Pinus spp.: Pinus nigra (3 samples), Pinus stankewiczii, Pinus brutia, Pinus halepensis, Pinus canariensis, Pinus pinaster and Pinus strobus from Greece were determined by gas chromatography and gas chromatography-mass spectrometry. P. nigra and P. canariensis essential oils were dominated by α-pinene (24.9-28.9 % and 15 %, respectively) and germacrene D (20.3-31.9 % and 55.8 %, respectively), whereas P. brutia and P. strobus by α-pinene (20.6 % and 31.4 %, respectively) and ß-pinene (31.7 % and 33.6 %, respectively). P. halepensis and P. pinaster oils were characterized by ß-caryophyllene (28.5 % and 22.5 %, respectively). Finally, ß-pinene (31.4 %), germacrene D (23.3 %) and α-pinene (17.5 %) were the most abundant compounds in the needle oil of P. stankewiczii. Additionally the larvicidal and repellent properties of their essential oils were evaluated against Aedes albopictus, a mosquito of great ecological and medical importance. The results of bioassays revealed that repellent abilities of the tested essential oils were more potent than their larvicidal activities. The essential oils of P. brutia, P. halepensis and P. stankewiczii presented considerable larvicidal activity (LC50 values 67.04 mgL(-1) and 70.21 mgL(-1), respectively), while the others were weak to inactive against larvae. The essential oils of P. halepensis, P. brutia, and P. stankewiczii presented a high repellent activity, even at the dose of 0.2 µL cm(-2), while in the dose of 0.4 µL cm(-2), almost all the tested EOs displayed protection against the mosquito.

Overwintering West Nile virus in active Culex pipiens mosquito populations in Greece.

The flavivirus West Nile Virus (WNV), which is transmitted by mosquitoes, poses a significant threat to both humans and animals, and its outbreaks often challenge public health in Europe and other continents. In recent years, there is an increasing trend of WNV incidence rates across several European countries. However, whether there is a year-round circulation or seasonal introduction has yet to be elucidated. Real-time polymerase chain reaction (PCR) identified WNV-positive Culex pipiens mosquitos in 6 out of 146 pools examined in winter 2022 that correspond to three out of the 24 study areas, located in two coastal regions units in Attica, Greece. Spatial dispersion of the six positive pools in the same region suggests a clustered circulation of WNV during the winter of 2022. This is the first study that documents the identification of WNV in Cx. pipiens populations, captured in adult traps during winter period. Our findings underscore the need to extend entomological surveillance programs to include the winter period, specifically in temperate climates and historically affected areas by WNV.

Sterile Insect Technique (SIT) field trial targeting the suppression of Aedes albopictus in Greece.

The sterile insect technique (SIT) involves releasing large numbers of sterile males to outcompete wild males in mating with females, leading to a decline in pest populations. In the current study, we conducted a suppression trial in Greece against the invasive dengue vector mosquito Aedes albopictus (Skuse) through the weekly release of sterile males for 22 weeks from June to September 2019. Our approach included the long-distance transport of sterile mosquitoes, and their release at a density of 2,547 ± 159 sterile males per hectare per week as part of an area-wide integrated pest management strategy (AW-IPM). The repeated releases of sterile males resulted in a gradual reduction in egg density, reaching 78% from mid-June to early September. This reduction remained between 70% and 78% for four weeks after the end of the releases. Additionally, in the SIT intervention area, the ovitrap index, representing the percentage of traps containing eggs, remained lower throughout the trial than in the control area. This trial represents a significant advance in the field of mosquito control, as it explores the viability and efficacy of producing and transporting sterile males from a distant facility to the release area. Our results provide valuable insights for future SIT programmes targeting Ae. Albopictus, and the methodology we employed can serve as a starting point for developing more refined and effective release protocols, including the transportation of sterile males over long distances from production units to intervention areas.

Title: Essai sur le terrain de la Technique de l'Insecte Stérile (TIS) ciblant la suppression d'Aedes albopictus en Grèce. Abstract: La technique de l'insecte stérile (TIS) consiste à libérer un grand nombre de mâles stériles pour supplanter les mâles sauvages lors de l'accouplement avec les femelles, entraînant ainsi un déclin des populations de nuisibles. Dans la présente étude, nous avons mené un essai de suppression en Grèce contre le moustique vecteur invasif de la dengue, Aedes albopictus (Skuse), par le biais de la libération hebdomadaire de mâles stériles pendant 22 semaines de juin à septembre 2019. Notre approche comprenait le transport sur de longues distances de moustiques stériles, et leur lâcher à une densité de 2 547 ± 159 mâles stériles par hectare et par semaine dans le cadre d'une stratégie de lutte intégrée contre les nuisibles à l'échelle de la zone (AW-IPM). Les lâchers répétés de mâles stériles ont entraîné une réduction progressive de la densité des œufs, atteignant 78 % de la mi-juin au début septembre. Cette réduction est restée entre 70 % et 78 % pendant quatre semaines après la fin des lâchers. De plus, dans la zone d'intervention de la TIS, l'indice d'oviposition, représentant le pourcentage de pièges contenant des œufs, est resté plus faible que dans la zone témoin tout au long de l'essai. Cet essai représente une avancée significative dans le domaine de la lutte contre les moustiques, car il explore la viabilité et l'efficacité de la production et du transport de mâles stériles depuis une installation éloignée vers la zone de lâcher. Nos résultats fournissent des informations précieuses pour les futurs programmes de TIS ciblant Ae. albopictus et la méthodologie que nous avons utilisée pourra servir de point de départ pour développer des protocoles de libération plus raffinés et plus efficaces, y compris le transport de mâles stériles sur de longues distances depuis les unités de production jusqu'aux zones d'intervention.

Pest categorisation of Cenopalpus irani.

The EFSA Panel on Plant Health performed a pest categorisation of Cenopalpus irani (Trombidiformes: Tenuipalpidae), known as the Iranian false spider mite, following the commodity risk assessment of Malus domestica plants from Türkiye, in which C. irani was identified as a pest of possible concern for the territory of the European Union (EU). The pest is only known to be present in Iran and Türkiye and has not been reported from the EU. The mite primarily feeds on Rosaceae plants but is considered polyphagous. Important crops of the EU that are hosts of C. irani include apples (Malus domestica), pears (Pyrus communis) and figs (Ficus carica). Plants for planting and fruits provide potential pathways for entry into the EU. Host availability and climate suitability in southern EU countries would most probably allow this species to successfully establish and spread. This mite is not listed in Annex II of Commission Implementing Regulation (EU) 2019/2072. Phytosanitary measures are available to reduce the likelihood of entry and spread of this species into the EU. The mite C. irani satisfies the criteria that are within the remit of EFSA to assess for it to be regarded as a potential Union quarantine pest, although there is a key uncertainty over the likelihood and magnitude of impact.

Pest categorisation of Crisicoccus seruratus.

Following the commodity risk assessments of Acer palmatum plants grafted on A. davidii from China, in which Crisicoccus matsumotoi (Hemiptera: Pseudococcidae) was identified as a pest of possible concern, the European Commission requested the EFSA Panel on Plant Health to conduct a pest categorisation of C. matsumotoi for the territory of the European Union. Recent taxonomic revision of the genus Crisisoccus concluded that C. matsumotoi is a synonym of C. seruratus; therefore, the categorisation will use the current valid name C. seruratus. It is an insect pest native to Japan, feeding on species in 13 plant families. There are reports of its presence also in China and the Republic of Korea, but there is great uncertainty about the identity of the species for these records. Therefore, there is uncertainty about the species referred to as C. matsumotoi in the commodity risk assessments of A. palmatum. C. seruratus is a multivoltine species. It has three generations per year and overwinters as a nymph. The most important crops that may be affected by C. seruratus are figs (Ficus carica), grapes (Vitis spp.), nashi pears (Pyrus pyrifolia var. culta), persimmons (Diospyros kaki) and walnuts (Juglans regia). Plants for planting and fruits provide potential pathways for entry into the EU. Host availability and climate suitability suggest that the central, northern and some areas of southern EU countries would be suitable for the establishment of C. seruratus. The introduction of this mealybug would likely have an economic impact in the EU through yield reduction and fruit downgrading because of honeydew deposition and the consequent growth of sooty moulds. This insect is not listed in Annex II of Commission Implementing Regulation (EU) 2019/2072. Phytosanitary measures are available to reduce the likelihood of entry and spread of this species into the EU. C. seruratus satisfies the criteria that are within the remit of EFSA to assess for it to be regarded as a potential Union quarantine pest.

Pest categorisation of Eulecanium giganteum.

The EFSA Panel on Plant Health performed a pest categorisation of Eulecanium giganteum (Hemiptera: Coccidae), the giant eulecanium scale, for the territory of the European Union, following the commodity risk assessment of Acer palmatum plants from China, in which E. giganteum came to attention as a pest of possible concern. The pest is only known to be present in Asia, where it has been reported from China, India, Iran, Japan and eastern Russia (Primorsky Krai). The pest has not been reported within the EU. It is not listed in Annex II of Commission Implementing Regulation (EU) 2019/2072. It is polyphagous, feeding on broad-leaf trees and shrubs assigned to 41 genera in 22 plant families. Host plant species commonly found in the EU include apricot (Prunus armeniaca), elm (Ulmus spp.), grapevine (Vitis vinifera), maple (Acer spp.), oak (Quercus spp.), oriental plane (Platanus orientalis), pomegranate (Punica granatum), quince (Cydonia oblonga), silkworm mulberry (Morus alba), walnut (Juglans regia), and several ornamentals. Climatic conditions and availability of host plants in southern EU countries would most probably allow this species to successfully establish and spread. However, EU native natural enemies are anticipated to provide biological control and therefore reduce potential impacts. Phytosanitary measures are available to reduce the likelihood of entry and spread. E. giganteum satisfies all the criteria that are within the remit of EFSA to assess for it to be regarded as a potential Union quarantine pest, other than the criterion on impact which is a key uncertainty.

Pest categorisation of Diaphania indica.

The EFSA Panel on Plant Health performed a pest categorisation of Diaphania indica (Lepidoptera: Crambidae), the cucumber moth for the territory of the European Union (EU), following the commodity risk assessment of Jasminum polyanthum from Uganda, in which D. indica was identified as a pest of possible concern to the European Union. D. indica is native to South Asian countries and is now distributed in tropical and subtropical areas of the Americas, Africa, Asia and Oceania. In the EU, D. indica occurs in Madeira (Portugal). It is a polyphagous pest, feeding on 16 genera in 6 plant families, primarily on plants of the Cucurbitaceae family. Important cucurbit hosts in the EU include cucumber (Cucumis sativus), melon (Cucumis melo), pumpkin (Cucurbita moschata), summer squash (Cucurbita pepo) and watermelon (Citrullus lanatus). Plants for planting, fruits and cut flowers provide potential pathways for entry into the EU. Climatic conditions and availability of host plants in southern EU countries would most probably allow this species to successfully establish and spread. Establishment could also occur in greenhouses in the northern parts of the EU. Economic impact in cultivated hosts, especially cucurbit crops is anticipated if establishment occurs. This insect is not listed in Annex II of Commission Implementing Regulation (EU) 2019/2072. Phytosanitary measures are available to reduce the likelihood of entry and further spread. D. indica meets the criteria that are within the remit of EFSA to assess for this species to be regarded as a potential Union quarantine pest.

Pest categorisation of Lepidosaphes malicola.

The EFSA Panel on Plant Health performed a pest categorisation of Lepidosaphes malicola (Hemiptera: Diaspididae), the Armenian mussel scale, for the territory of the European Union, following commodity risk assessments of Prunus persica and P. dulcis plants for planting from Türkiye, in which L. malicola was identified as a pest of possible concern. L. malicola is a polyphagous insect of temperate and arid areas, feeding on more than 60 plant species belonging to 26 families. Important crops significantly affected by L. malicola in parts of Asia include stone fruits (Prunus armeniaca, P. persica), pome fruits (Malus domestica, Pyrus communis), grapes (Vitis vinifera), pomegranate (Punica granatum), walnuts (Juglans regia) and ornamental plants (Berberis spp., Cornus spp., Jasminum spp., Ligustrum spp.). L. malicola has two generations annually. The overwintered eggs hatch from late May to early June. First-instar nymphs crawl on the host plant for a short period, then settle to feed. Nymphs reach maturity in late summer or early autumn. Plants for planting, fruits and cut flowers provide potential pathways for entry into the EU. Host availability and climate suitability suggest that southern, central and some parts of northern EU countries would be suitable for the establishment of L. malicola. Despite being a pest in Armenia, Iran and Tajikistan, there is no evidence of it being a pest in Türkiye. L. malicola was detected in Bulgaria and Greece over 30 years ago, but there have been no records since, and its status is uncertain. Its ability to cause an impact in the EU is also uncertain. It is not listed in Annex II of the Commission Implementing Regulation (EU) 2019/2072. Phytosanitary measures are available to reduce the likelihood of entry. Except for the criterion of having an economic or environmental impact, for which there is great uncertainty, L. malicola satisfies all other criteria that are within the remit of EFSA to assess for it to be regarded as a potential Union quarantine pest.

Commodity risk assessment of Cornus alba and Cornus sanguinea plants from the UK.

The European Commission requested the EFSA Panel on Plant Health to prepare and deliver risk assessments for commodities listed in Commission Implementing Regulation (EU) 2018/2019 as 'high risk plants, plant products and other objects'. Taking into account the available scientific information, including the technical information provided by the applicant country, this Scientific Opinion covers the plant health risks posed by the following commodities: Cornus alba and Cornus sanguinea bare-root plants and rooted plants in pots up to 7 years old imported into the EU from the UK. A list of pests potentially associated with the commodities was compiled. The relevance of any pest was assessed based on evidence following defined criteria. Four EU quarantine pests (Meloidogyne fallax, Phytophthora ramorum (non-EU isolates), tobacco ringspot virus, and tomato ringspot virus) and one EU non-regulated pest (Discula destructiva), were selected for further evaluation. For the selected pests, the risk mitigation measures implemented in the technical dossier from the UK were evaluated taking into account the possible limiting factors. For these pests, an expert judgement is given on the likelihood of pest freedom taking into consideration the risk mitigation measures acting on the pest, including uncertainties associated with the assessment. The degree of pest freedom varies among the pests evaluated, with P. ramorum being the pest most frequently expected on the imported C. alba and C. sanguinea plants. The Expert Knowledge Elicitation indicated, with 95% certainty, that between 9823 and 10,000 bare-root C. alba and C. sanguinea plants per 10,000 will be free from P. ramorum.

Commodity risk assessment of Sorbus aucuparia plants from the UK.

The European Commission requested the EFSA Panel on Plant Health to prepare and deliver risk assessments for commodities listed in Commission Implementing Regulation (EU) 2018/2019 as 'high risk plants, plant products and other objects'. Taking into account the available scientific information, including the technical information provided by the applicant country, this Scientific Opinion covers the plant health risks posed by the following commodities: Sorbus aucuparia bare-root plants and rooted plants in pots up to 7 years old and specimen trees in pots up to 15 years old imported into the EU from the UK. A list of pests potentially associated with the commodities was compiled. The relevance of any pest was assessed based on evidence following defined criteria. Three EU quarantine pests (Entoleuca mammata and Phytophthora ramorum (non-EU isolates), Erwinia amylovora), were selected for further evaluation. For two of the selected pests (E. mammata and P. ramorum), the risk mitigation measures implemented in the UK and specified in the technical dossier were evaluated taking into account the possible limiting factors. For these pests, an expert judgement is given on the likelihood of pest freedom taking into consideration the risk mitigation measures acting on the pest, including uncertainties associated with the assessment. The degree of pest freedom varies between the pests evaluated, with P. ramorum being the pest most frequently expected on the imported S. aucuparia plants. The Expert Knowledge Elicitation indicated, with 95% certainty, that between 9812 and 10,000 bare-root S. aucuparia plants per 10,000 will be free from P. ramorum.

Commodity risk assessment of Petunia spp. and Calibrachoa spp. unrooted cuttings from Guatemala.

The European Commission requested the EFSA Panel on Plant Health to evaluate the probability of entry of pests (likelihood of pest freedom at entry), including both, regulated and non-regulated pests, associated with unrooted cuttings of the genera Petunia and Calibrachoa produced under physical isolation in Guatemala. The relevance of any pest for this opinion was based on evidence following defined criteria, based on the methodology used for high-risk plants adapted for the specificity of this assessment. Nineteen EU regulated pests (Bemisia tabaci, pepper golden mosaic virus, pepper huasteco yellow vein virus, tomato severe leaf curl virus, tomato yellow leaf curl virus, tomato spotted wilt virus, Liriomyza huidobrensis, Liriomyza sativae, Liriomyza trifolii, Bactericera cockerelli, Eotetranichus lewisi, Epitrix subcrinita, Epitrix cucumeris, Helicoverpa zea, Chloridea virescens, Spodoptera ornithogalli, Ralstonia solanacearum, Ralstonia pseudosolanacearum, Xanthomonas vesicatoria) and one EU non-regulated (Phenacoccus solenopsis) pest fulfilled all relevant criteria and were selected for further evaluation. For these pests, the risk mitigation measures proposed in the technical dossier from Guatemala were evaluated taking into account the possible limiting factors, and an expert judgement is given on the likelihood of pest freedom taking into consideration the risk mitigation measures acting on the pest, including uncertainties associated with the assessment. The limited and partially conflicting information provided in the dossier contributes to the wide estimates of pest freedom. The estimated degree of pest freedom varies among the pests evaluated, with Ralstonia spp. (R. solanacearum and R. pseudosolanacearum) being the pest most frequently expected on the imported cuttings. The expert knowledge elicitation indicated, with 95% certainty, that between 9916 and 10,000 bags containing unrooted cuttings per 10,000 would be free of Ralstonia spp.

Commodity risk assessment of Petunia spp. and Calibrachoa spp. unrooted cuttings from Kenya.

The European Commission requested the EFSA Panel on Plant Health to evaluate the probability of entry of pests (likelihood of pest freedom at entry), including both regulated and non-regulated pests, associated with unrooted cuttings of the genera Petunia and Calibrachoa produced under physical isolation in Kenya. The relevance of any pest for this opinion was based on evidence following defined criteria, based on the methodology used for High-Risk Plants adapted for the specificity of this assessment. Fourteen EU-regulated pests (Bemisia tabaci, cowpea mild mottle virus, Liriomyza huidobrensis, Liriomyza sativae, Liriomyza trifolii, potato leafroll virus, potato spindle tuber viroid, Ralstonia pseudosolanacearum, R. solanacearum, Scirtothrips dorsalis, tomato mild mottle virus, tomato spotted wilt virus, tomato yellow leaf curl virus and Xanthomonas vesicatoria) and six EU non-regulated pests (Aleurodicus dispersus, pepper veinal mottle virus, Nipaecoccus viridis, Phenacoccus solenopsis, Tetranychus neocaledonicus and tomato yellow ring virus) fulfilled all relevant criteria and were selected for further evaluation. For these pests, the risk mitigation measures proposed in the technical dossier from Kenya were evaluated, taking into account the possible limiting factors. Additionally, an expert judgement is given on the likelihood of pest freedom, taking into consideration the risk mitigation measures acting on the pest, including uncertainties associated with the assessment. The estimated degree of pest freedom varies among the pests evaluated, with T. neocaledonicus being the pest most frequently expected on the imported cuttings. The Expert Knowledge Elicitation indicated, with 95% certainty, that between 9942 and 10,000 bags containing unrooted cuttings of Petunia spp. and Calibrachoa spp. per 10,000 would be free of T. neocaledonicus.

Essential oil composition, adult repellency and larvicidal activity of eight Cupressaceae species from Greece against Aedes albopictus (Diptera: Culicidae).

The present study evaluated leaf essential oils from eight Cupresaceae species; Cupressus arizonica, Cupressus benthamii, Cupressus macrocarpa, Cupressus sempervirens, Cupressus torulosa, Chamaecyparis lawsoniana, Juniperus phoenicea, and Tetraclinis articulata for their larvicidal and repellent properties against Aedes albopictus, a mosquito of great ecological and medical importance. Based on the LC(50) values, C. benthamii essential oil was the most active (LC(50) = 37.5 mg/L) while the other tested Cupressaceae essential oils provided rather moderate toxicity against larvae (LC(50) = 47.9 to 70.6 mg/L). Under the used laboratory conditions, three of the essential oils (C. benthamii, C. lawsoniana, and C. macrocarpa) provided sufficient protection against mosquito adults, equivalent to the standard repellent "Deet" in the 0.2 mg/cm(2) dose, while C. macrocarpa assigned as the superior repellent oil in the 0.08 mg/cm(2) dose. Chemical analysis of the essential oils using gas chromatography and gas chromatography-mass spectrometry revealed the presence of 125 components.

Insecticidal activity of plant essential oils against the vine mealybug, Planococcus ficus.

The vine mealybug, Planococcus ficus (Signoret) (Hemiptera: Pseudococcidae), is a pest in grape vine growing areas worldwide. The essential oils from the following aromatic plants were tested for their insecticidal activity against P. ficus: peppermint, Mentha piperita L. (Lamiales: Lamiaceae), thyme-leaved savory, Satureja thymbra L., lavender, Lavandula angustifolia Mill, and basil, Ocimum basilicum L. Essential oils from peels of the following fruits were also tested: lemon, Citrus limon L. (Sapindales: Rutaceae), and orange, C. sinensis L. The reference product was paraffin oil. Bioassays were conducted in the laboratory by using spray applications on grape leaves bearing clusters of P. ficus of one size class, which mainly represented either 3rd instar nymphs or pre-ovipositing adult females. The LC50 values for each essential oil varied depending on the P. ficus life stage but did not significantly differ between 3(rd) instar nymphs and adult females. The LC50 values of the citrus, peppermint, and thyme-leaved savory essential oils ranged from 2.7 to 8.1 mg/mL, and the LC50 values of lavender and basil oil ranged from 19.8 to 22.5 and 44.1 to 46.8 mg/mL, respectively. The essential oils from citrus, peppermint and thymeleaved savory were more or equally toxic compared to the reference product, whereas the lavender and basil essential oils were less toxic than the paraffin oil. No phytotoxic symptoms were observed on grape leaves treated with the citrus essential oils, and low phytotoxicity was caused by the essential oils of lavender, thyme-leaved savory, and mint, whereas the highest phytotoxicity was observed when basil oil was used.