Laboratory experiments were conducted to study the bioefficacy against Ae. albopictus of three Citrus essential oils, derived from peels of Citrus sinensis, Citrus limon, and Citrus paradise and of their components. Chiral gas chromatography analysis revealed the dominant occurrence of R-(+)-limonene and (-)-β-pinene in all three essential oils while in the case of lemon oil γ-terpinene, neral, and geranial detected also among other components. The tested Citrus essential oils were toxic against mosquito larvae with LC(50) values ranging from 25.03 to 37.03 mg l(-1). Among citrus essential oils components tested, γ-terpinene was the most toxic (LC(50) = 20.21 mg l(-1)) followed by both enantiomeric forms of limonene (LC(50) = 35.99 and 34.89 mg l(-1), for R-(+)-limonene and S-(-)-limonene, respectively). The delayed toxic effects after exposure of larvae to sublethal (LC(50)) doses were also investigated for citrus essential oils and their major component R-(+)-limonene, indicating a significant reduction of pupal survival. In repellent bioassays, lemon essential oil, S-(-)-limonene, citral (mixture of neral\geranial) and (+)-β-pinene were the most effective compared with other citrus essential oils and components against adult mosquitoes. Repellent bioassays also revealed that limonenes and β-pinenes showed an isomer dependence repellent activity. Finally, according to enantiomeric distribution of limonene and α- and β-pinene, the repellency of lemon essential oil is possibly attributed to the presence of citral.

The volatile metabolites of wild-growing Mentha spicata, M. longifolia, M. suaveolens, Melissa officinalis, Salvia fruticosa, S. pomifera subsp. calycina, and S. pomifera subsp. pomifera from Greece were determined by gas chromatography and gas chromatography-mass spectrometry. The insecticidal properties of the analyzed essential oils were screened on Culex pipiens larvae. Additionally two of the main components of the essential oils, piperitenone oxide and 1,8-cineole were assayed against C. pipiens in order to define the affiliation between them and the larvicidal properties of the oils. The most effective oils were M. suaveolens (major constituent piperitenone oxide, 62.4%), M. spicata (piperitenone oxide, 35.7% and 1,8-cineole, 14.5%) and M. longifolia--Central Greece (piperitenone oxide, 33.4%; 1,8-cineole, 24.5% and trans-piperitone epoxide, 17.4%), which exhibited LC(50) values ranging from 47.88 to 59.33 mg l(-1). Medium activity revealed the oils of M. officinalis (terpin-4-ol, 15.8%; caryophyllene oxide, 13.2%; sabinene, 12.9%; beta-pinene, 12.1%; and trans-caryophyllene, 10.2%), M. longifolia--Southern Greece (carvone, 54.7% and limonene 20.0%), S. pomifera subsp. pomifera (trans-caryophyllene, 22.5% and trans-thujone, 21.0%), S. pomifera subsp. calycina--West Southern Greece (trans-thujone, 56.1% and 1,8-cineole, 10.4%), and S. fruticosa--population 2 (camphor, 23.1%; alpha-pinene, 12.7%; and borneol, 12.6%), with LC(50) values ranging from 78.28 to 91.45 mg l(-1). S. pomifera subsp. calycina (Central Greece) essential oil (trans-thujone, 26.5% and cis-thujone, 12.0%) presented rather low activity (LC(50) values 140.42 mg l(-1)), while S. fruticosa--population 1 (1,8-cineole, 31.4% and camphor, 22.6%) was the only inactive oil. Additionally, the constituent piperitenone oxide was found to be highly active (LC(50) values 9.95 mg l(-1)), whereas 1,8-cineole revealed no toxicity.

The aim of this study was to evaluate the toxicity of pinenes (enantiomers of alpha- and beta-) and essential oils from Greek plants of the Rutaceae family against the mosquito larvae of Culex pipiens (Diptera: Culicidae). Essential oils were isolated by hydrodistillation from fruit peel of orange (Citrus sinensis L.), lemon (Citrus limon L.), and bitter orange (Citrus aurantium L.). The chemical composition was determined by gas chromatography/mass spectrometry (GC/MS) analysis. Citrus essential oils contained in high proportion limonene and in lower quantities p-menthane molecules and pinenes. The insecticidal action of these essential oils and enantiomers of their pinenes on mosquito larvae was evaluated. Plant essential oils exhibited strong toxicity against larvae with the LC(50) values ranging from 30.1 (lemon) to 51.5 mg/L (orange) depending on Citrus species and their composition. Finally, the LC(50) value of pinenes ranging from 36.53 to 66.52 mg/L indicated an enantioselective toxicity only for the beta-pinene enantiomer.

The addition of DDS to EO did not change the toxic effect of the essential oil, whereas the presence of DS in excess produced an enriched essential oil with low toxicity. Furthermore, the antagonistic effect of DS and DDS against the other components of EO was shown by the application of two equations obtained from the literature. Their larvicidal performances were correlated, for the first time, with their detailed chemical composition. Data analysis strongly indicated the toxicity of the other EO sulfur ingredients.

Background: Human cases of West Nile virus (WNV) infection are recorded since 2010 in Greece, with seasonal outbreaks occurring almost annually. Enhanced surveillance has been implemented since 2010, to promptly characterise cases' temporal and geographical distribution and inform authorities for implementation of appropriate measures (mosquito control, health education, blood safety). Aim: We describe the epidemiology of WNV human infections in Greece focusing on the 2018 season. Methods: The National Public Health Organization advised physicians to test all suspect WNV infection cases and refer samples to reference laboratories. Laboratories notified diagnosed cases on a daily basis. Treating physicians, patients, and infected blood donors were interviewed within 48 hours after diagnosis and the probable infection location was identified. Hospitalised cases were followed up until discharge. Results: A total of 317 autochthonous WNV infection cases were diagnosed in 2018. Among them, 243 cases had neuroinvasive disease (WNND), representing a 23% increase of WNND cases compared with 2010, the previous most intense season. There were 51 deaths. Cases started occurring from week 22, earlier than usual. Both rural and urban areas were affected, with 86 (26% of the total) municipalities belonging to seven (54% of the total) regions recording cases. Two major epicentres were identified in Attica and Central Macedonia regions. Conclusions: The largest number of human cases of WNV infection ever recorded in Greece occurred in 2018, with a wide geographical distribution, suggesting intense virus circulation. Enhanced surveillance is vital for the early detection of human cases and the prompt implementation of response measures.