Volatile oils of Thymus serpyllum and Artemisia absinthium: GS-MS analysis and insecticidal activity against Culiseta longiareolata

F. Aouir; R. Chaibi; B. Merabti; S. Benhissen; I. Sifi; H. Gouzi

doi:10.15421/012513

F. Aouir Amar Telidji University
R. Chaibi Amar Telidji University
B. Merabti Mohamed Kheider University
S. Benhissen Mohamed Boudiaf University
I. Sifi Amar Telidji University
H. Gouzi Amar Telidji University

Keywords: essential oil; Asteraceae; Lamiaceae; Culicidae; larvicidal activity; mosquitoes; Algeria

Abstract

Culiseta longiareolata (Macquart, 1838) (Diptera, Culicidae) is one of the important mosquito species distributed in Algeria. It acts as a vector of avian malaria, tularemia, Malta fever (brucellosis), and several arboviruses, such as West Nile fever and western encephalitis virus. To eliminate undesirable insect vectors of pathogens, botanical-based insecticides are increasingly recommended. This research was focused on the analysis of the chemical composition of essential oils from Artemisia absinthium (Asteraceae) and Thymus serpyllum (Lamiaceae) obtained by hydrodistillation using gas chromatography – mass spectroscopy (GC-MS). Furthermore, these oils were tested for the first time for their larvicidal activity against fourth-instar larvae of C. longiareolata. In the Th. serpyllum essential oil, linalool (25.40%) was the major constituent, followed by camphor (19.75%) and α-pinene (11.25%). In the A. absinthium essential oil, the major compound was camphor (39.01%), followed by chamazulene (14.51%). Our study found significant larvicidal effects against the fourth-instar larvae, with a direct correlation among the larval mortality rate, applied concentrations, and duration of exposure to the treatment, where the mortality increased over time. Also, the results indicated a significant difference in the mortality rate between the two samples of essential oils. The essential oil extracted from A. absinthium was highly effective against mosquito larvae, achieving the LC50 values of 10.42, 6.19 and 3.82 ppm at 24, 48, and 72 h, respectively, while the essential oil from Th. serpyllum achieved the LC50 values of 12.16, 7.79, and 6.88 ppm. Based on the current results, both the plants could be used as new alternatives for mosquito control.

References

Adams, R. P. (2017). Identification of essential oil components by gas chromatography/mass spectrometry. Texensis Publishing, Gruver. Pp. 46–52.

Ahmad, A. M., Khokhar, I., Ahmad, I., Kashmiri, M. A., Adnan, A., & Ahmad, M. (2006). Study of antimicrobial activity and composition by GC/MS spectroscopic analysis of the essential oil of Thymus serpyllum. Internet Journal of Food Safety, 5, 56–60.

Alami, A., Lalami, A. E. O., Annemer, S., El‐Akhal, F., Ez‐zoubi, A., El Hallabi, M., ... & Farah, A. (2024). Larvicidal activity of six Moroccan Artemisia essential oils and a correlation study of their major components against Culex pipiens. Journal of Applied Entomology, 148(8), 948–964.‏

Alayat, M. S., Bendali-Saoudi, F., Mahmoudi, K., & Soltani, N. (2023). Diversity and spatio-temporal distribution of mosquitoes (Diptera: Culicidae) in the Laghouat arid region (Algerian Northern Sahara). Oriental Insects, 57(4), 1102–1127.‏

Al-Ghamdi, A. A. (2020). Ecological and biochemical studies on Artemisia absinthium in Al-Baha City, Saudi Arabia. Pakistan Journal of Botany, 52, 1421–1425.‏

Ali, S. I., Gopalakrishnan, B., & Venkatesalu, V. (2018). Chicory (Cichorium intybus) and wormwood (Artemisia absinthium) extracts exhibit strong larvicidal activity against mosquito vectors of malaria, dengue fever, and filariasis. Parasitology International, 67(6), 781–786.

Amer, A., & Mehlhorn, H. (2006). Larvicidal effects of various essential oils against Aedes, Anopheles, and Culex larvae (Diptera, Culicidae). Parasitology Research, 99, 466–472.

Asfaw, N., & Demissew, S. (2015). Essential oil composition of four Artemisia species from Ethiopia. Bulletin of the Chemical Society of Ethiopia, 29(1), 123–128. ‏

Baba Aissa, F. (1991). Les plantes médicinales en Algérie [Medicinal plants in Algeria]. Coédition Bouchene et ad, Diwan Alger (in French).

Babushok, V. I., Linstrom, P. J., & Zenkevich, I. G. (2011). Retention indices for frequently reported compounds of plant essential oils. Journal of Physical and Chemical Reference Data, 40(4), 043101.

‏Baj, T., Biernasiuk, A., Wróbel, R., & Malm, A. (2020). Chemical composition and in vitro activity of Origanum vulgare L., Satureja hortensis L., Thymus serpyllum L. and Thymus vulgaris L. essential oils towards oral isolates of Candida albicans and Candida glabrata. Open Chemistry, 18(1), 108–118.

‏Bencheqroun, H. K., Ghanmi, M., Satrani, B., Aafi, A., & Chaouch, A. (2012). Activité antimicrobienne des huiles essentielles d’Artemisia mesatlantica, plante endémique du Maroc [Antimicrobial activity of the essential oil of an endemic plant in Morocco Artemisia mesatlantica]. Bulletin de la Société Royale des Sciences de Liège ‏(in French).

Benhissen, S., Asloum, A. Y., Belkhiri, N., Hedjouli, Z., Bounadji, S., Habbachi, W., & Rebbas, K. (2024). Toxicity of three species of the Solanaceae family growing in Algeria against Culiseta longiareolata mosquitos 4th stage larvae. Journal of Bioresource Management, 11(1), 1.‏

Benkhaled, A., Boudjelal, A., Napoli, E., Baali, F., & Ruberto, G. (2020). Phytochemical profile, antioxidant activity and wound healing properties of Artemisia absinthium essential oil. Asian Pacific Journal of Tropical Biomedicine, 10(11), 496–504.‏

Bertouche, S., Sabba, N., Sahraoui, N., & Bachir, Y. N. (2023). Extraction of Thymus pallecens de Noé essential oil by microwave steam distillation and steam distillation processes: Kinetics, optimization, chemical composition and insecticidal activity. Algerian Journal of Environmental Science and Technology, 9(1), 3026–3036.‏

Blagojević, P., Radulović, N., Palić, R., & Stojanović, G. (2006). Chemical composition of the essential oils of Serbian wild-growing Artemisia absinthium and Artemisia vulgaris. Journal of Agricultural and Food Chemistry, 54(13), 4780–4789.‏

Bouabida, H., & Dris, D. (2020). Effect of rue (Ruta graveolens) essential oil on mortality, development, biochemical and biomarkers of Culiseta longiareolata. South African Journal of Botany, 133, 139–143.‏

Bouaziz, A., Aissaoui, L., & Boudjelida, H. (2023). The activity of Bacillus thuringiensis Vectobac G on the biochemical compositions of Culex pipiens and Culiseta longiareolata (Diptera: Culicidae) mosquito larvae. Ecology, Environment and Conservation Journal, 29(1), 1–9.‏

Bouchenak, F., Degaichia, H., Lamgharbi, A., & Benrebiha, F. (2018). Evaluation in vitro du potentiel antifongique de l’huile essentielle et des extraits méthanoliques d’une asteraceae Artemisia absinthium L. [In vitro evaluation of the antifungal potential of the essential oil and methanolic extracts of an asteraceae Artemisia absinthium L.]. Agrobiologia, 8(1), 886–895 ‏(in French).

Bouguerra, N., Tine-Djebbar, F., & Soltani, N. (2018). Effect of Thymus vulgaris L.(Lamiales: Lamiaceae) essential oil on energy reserves and biomarkers in Culex pipiens L.(Diptera: Culicidae) from Tebessa (Algeria). Journal of Essential oil Bearing Plants, 21(4), 1082–1095.‏

Boumaza, M., Merabti, B., Adjami, Y., Ouakid, M. L., & Carvajal, T. M. (2022). Geometric morphometric wing analysis of avian malaria vector, Culiseta longiareolata, from two locations in Algeria. Insects, 13(11), 1031.‏

Bounadji, S., Benhissen, S., Belkhiri, N., Asloum, A.Y., Habbachi, W., & Rebbas, K. H. (2024). Biological struggle against two harmful insects Blattella germanica (Blattellidae) and Culiseta longiareolata (Culicidae) using Euphorbia bupleuroides (Euphorbiaceae). Sustainable Development, 14(1), 11–20.

Bruneton, J. (2002). Elément de phytochimie et de pharmacognosie [Elements of phytochemistry and pharmacognosy]. 3ème édition, Lavoisier, Paris ‏(in French).

Brunhes, J., Rhaim, A., Geoffroy, B., Angel, G., & Hervy, J. P. (2000). Les moustiques de l’Afrique méditerranéenne: Logiciel d’identification et d’enseignement [Mosquitoes of Mediterranean Africa: Identification and teaching software]. Éditions IRD, Montpellier ‏(in French).

Chaouch, R., Bouaziz, A., Gacem, H., & Saoudi, B. (2024). Study of the toxic effect of chemical pesticide Actellic 50EC against Culiseta longiareolata mosquito larvae.‏ Indian Journal of Entomology, 86(2), 356–361.

Chebbac, K., Benziane Ouaritini, Z., El Moussaoui, A., Chalkha, M., Lafraxo, S., Bin Jardan, Y. A., ... & Guemmouh, R. (2023). Antimicrobial and antioxidant properties of chemically analyzed essential oil of Artemisia annua L. (Asteraceae) native to Mediterranean area. Life, 13(3), 807.‏

Chebbac, K., Ghneim, H. K., El Moussaoui, A., Bourhia, M., El Barnossi, A., Benziane Ouaritini, Z., ... & Guemmouh, R. (2022). Antioxidant and antimicrobial activities of chemically-characterized essential oil from Artemisia aragonensis Lam. against drug-resistant microbes. Molecules, 27(3), 1136.‏

Chebbac, K., Moussaoui, A. E., Bourhia, M., Salamatullah, A. M., Alzahrani, A., & Guemmouh, R. (2021). Chemical analysis and antioxidant and antimicrobial activity of essential oils from Artemisia negrei L. against drug-resistant microbes. Evidence‐Based Complementary and Alternative Medicine, 2021(1), 5902851.‏

Damtie, D., & Mekonnen, Y. (2021). Toxicity and oviposition deterrent activities of thyme essential oils against Anopheles arabiensis. Psyche, 2021(1), 6684156.‏

D'Auria, M., & Racioppi, R. (2015). The effect of drying of the composition of volatile organic compounds in Rosmarinus officinalis, Laurus nobilis, Salvia officinalis and Thymus serpyllum. A HS-SPME-GC-MS study. Journal of Essential Oil Bearing Plants, 18(5), 1209–1223.‏

Deepalakshmi, S., & Jeyabalan, D. (2017). Studies on mosquitocidal and biological activity of endemic plants of Nilgiris Hills against filarial vector, Culex quinquefasciatus (Say) (Insecta: Diptera: Culicidae). International Journal of Advanced Research in Biological Sciences, 4(3), 137–151. ‏

Derwich, E., Benziane, Z., & Boukir, A. (2009). Chemical compositions and insectisidal activity of essential oils of three plants Artemisia sp: Artemisia herba-alba, Artemisia absinthium and Artemisia pontica (Morocco). Electronic Journal of Environmental, Agricultural and Food Chemistry, 8(12), 1202–1211. ‏

De Lisi, A., Tedone, L., Montesano, V., Sarli, G., & Negro, D. (2011). Chemical characterisation of Thymus populations belonging from Southern Italy. Food Chemistry, 125(4), 1284–1286.‏

Djemâa, D., & Hayette, B. (2023). Assessement of larvicidal and pupicidal activities of Mentha piperita essential oil and effects in biomarkers and morphometric aspects against two mosquito species (Culiseta longiareolata and Culex pipiens). International Journal of Tropical Insect Science, 43(6), 1897–1909. ‏

Dris, D. (2018). Etude de l’activité larvicide des extraits de trois plantes: Mentha piperita, Lavandula dentata et Ocimum basilicum sur les larves de deux espèces de moustiques Culex pipiens (Linné) et Culiseta longiareolata (Aitken) [Study of the larvicidal activity of extracts of three plants: Mentha piperita, Lavandula dentata and Ocimum basilicum on the larvae of two species of mosquitoes Culex pipiens (Linnae) and Culiseta longiareolata (Aitken)]. Université de Badji mokhtar, Annaba, Algérie (in French).

Dris, D., Tine-Djebbar, F., & Soltani, N. (2017). Lavandula dentata essential oils: Chemical composition and larvicidal activity against Culiseta longiareolata and Culex pipiens (Diptera: Culicidae). African Entomology, 25(2), 387–394.‏

El Ouahdani, K., Es-Safi, I., Mechchate, H., Al-Zahrani, M., Qurtam, A. A., Aleissa, M., ... & Bousta, D. (2021). Thymus algeriensis and Artemisia herba-alba essential oils: Chemical analysis, antioxidant potential and in vivo anti-inflammatory, analgesic activities, and acute toxicity. Molecules, 26(22), 6780.‏

Eweis, M., Imhemmed, A. A. A., & Gad, A. S. (2012). Influence of Thymus serpyllum essential oil on Aspergillus parasiticus morphology and aflatoxins production. Research Journal of Pharmaceutical, Biological and Chemical Sciences, 3(2), 322–332.

Feng, Y. X., Zhang, X., Wang, Y., Chen, Z. Y., Lu, X. X., Du, Y. S., & Du, S. S. (2021). The potential contribution of cymene isomers to insecticidal and repellent activities of the essential oil from Alpinia zerumbet. International Biodeterioration and Biodegradation, 157, 105138.‏

Finney, D. J. (1971). Probit analysis. Cambridge University Press, Cambridge. ‏

Fuzimoto, A. D. (2021). An overview of the anti-SARS-CoV-2 properties of Artemisia annua, its antiviral action, protein-associated mechanisms, and repurposing for COVID-19 treatment. Journal of Integrative Medicine, 19(5), 375–388. ‏

Galovičová, L., Borotová, P., Valková, V., Vukovic, N. L., Vukic, M., Terentjeva, M., ... & Kačániová, M. (2021). Thymus serpyllum essential oil and its biological activity as a modern food preserver. Plants, 10(7), 1416. ‏

Govindarajan, M., & Benelli, G. (2016). Artemisia absinthium-borne compounds as novel larvicides: Effectiveness against six mosquito vectors and acute toxicity on non-target aquatic organisms. Parasitology Research, 115, 4649–4661. ‏

Goyal, S., Pathak, R., Pandey, H. K., Kumari, A., Tewari, G., Bhandari, N. S., & Bala, M. (2020). Comparative study of the volatile constituents of Thymus serpyllum L. grown at different altitudes of Western Himalayas. SN Applied Sciences, 2(7), 1208. ‏

Handa, S. S. (2008). An overview of extraction techniques for medicinal and aromatic plants. Extraction Technologies for Medicinal and Aromatic Plants, 1(1), 21–40. ‏

Jiang, C., Zhou, S., Liu, L., Toshmatov, Z., Huang, L., Shi, K., ... & Shao, H. (2021). Evaluation of the phytotoxic effect of the essential oil from Artemisia absinthium. Ecotoxicology and Environmental Safety, 226, 112856.

Joshi, R. K. (2013). Volatile composition and antimicrobial activity of the essential oil of Artemisia absinthium growing in Western Ghats region of North West Karnataka, India. Pharmaceutical Biology, 51(7), 888–892. ‏

Judzentiene, A., & Budiene, J. (2010). Compositional variation in essential oils of wild Artemisia absinthium from Lithuania. Journal of Essential Oil Bearing Plants, 13(3), 275–285. ‏

Judzentiene, A., Tomi, F., & Casanova, J. (2009). Analysis of essential oils of Artemisia absinthium L. from Lithuania by CC, GC (RI), GC-MS and 13C NMR. Natural Product Communications, 4(8), 1934578X0900400820. ‏

Khan, F. A., Khan, N. M., Ahmad, S., Nasruddin, Aziz, R., Ullah, I., ... & Aljuaid, A. (2022). Phytochemical profiling, antioxidant, antimicrobial and cholinesterase inhibitory effects of essential oils isolated from the leaves of Artemisia scoparia and Artemisia absinthium. Pharmaceuticals, 15(10), 1221. ‏

Kharoubi, R., Rehimi, N., Khaldi, R., Haouari-Abderrahim, J., & Soltani, N. (2021). Phytochemical screening and insecticidal activities of essential oil of Mentha x piperita L. (Lamiales: Lamiaceae) and their enzymatic properties against mosquito Culex pipiens L. (Diptera: Culicidae). Journal of Essential Oil Bearing Plants, 24(1), 134–146. ‏

Khebri, S. (2011). Etude chimique et biologique des huiles essentielles de trois Artemisia [Chemical and biological study of essential oils of three Artemisia]. Université El Hadj Lakhder, Batna (in French).

Kordali, S., Cakir, A., Mavi, A., Kilic, H., & Yildirim, A. (2005). Screening of chemical composition and antifungal and antioxidant activities of the essential oils from three Turkish Artemisia species. Journal of Agricultural and Food Chemistry, 53(5), 1408–1416. ‏

Kulišić, T., Kriško, A., Dragović-Uzelac, V., Miloš, M., & Pifat, G. (2007). The effects of essential oils and aqueous tea infusions of oregano (Origanum vulgare L. spp. hirtum), thyme (Thymus vulgaris L.) and wild thyme (Thymus serpyllum L.) on the copper-induced oxidation of human low-density lipoproteins. International Journal of Food Sciences and Nutrition, 58(2), 87–93. ‏

Lakhdari, W., Bouhenna, M. M., Neghmouche, N. S., Dehliz, A., Benyahia, I., Bendif, H., & Garzoli, S. (2024). Chemical composition and insecticidal activity of Artemisia absinthium L. essential oil against adults of Tenebrio molitor L. Biochemical Systematics and Ecology, 116, 104881.

Lamarti, A., Badoc, A., & Carde, J. P. (1993). Étude chromatographique de l'huile essentielle de la plantule de fenouil amer (Foeniculum vulgare Mill.); caractéristiques spectrales (uv, ir, sm) de ses constituants [Chromatographic study of the essential oil of bitter fennel seedling (Foeniculum vulgare Mill.); spectral characteristics (UV, IR, MS) of its constituents]. Bulletin de la Société de Pharmacie, 132, 73–89 (in French).

Li, X., He, T., Wang, X., Shen, M., Yan, X., Fan, S., ... & She, G. (2019). Traditional uses, chemical constituents and biological activities of plants from the genus Thymus. Chemistry and Biodiversity, 16(9), e1900254.

Lichfield, J. T., & Wilcoxon, F. A. (1949). A simplified method of evaluation doses-effects experiments. Journal of Pharmacology and Experimental Therapeutics, 95, 996–1113.

Luz, T. R. S. A., de Mesquita, L. S. S., do Amaral, F. M. M., & Coutinho, D. F. (2020). Essential oils and their chemical constituents against Aedes aegypti L. (Diptera: Culicidae) larvae. Acta Tropica, 212, 105705.

Maleš, I., Dragović-Uzelac, V., Jerković, I., Zorić, Z., Pedisić, S., Repajić, M., ... & Dobrinčić, A. (2022). Non-volatile and volatile bioactives of Salvia officinalis L., Thymus serpyllum L. and Laurus nobilis L. extracts with potential use in the development of functional beverages. Antioxidants, 11(6), 1140.

Mazulin, O. V., Fukleva, L. A., & Mazulin, G. V. (2023). Study of the polyphenolic compounds accumulation in Thymus serpyllum L. herb during the flowering period.‏ Pharmaceutical Journal, 78(2), 58–66.

Messara, Y., Fernane, F., & Meddour, R. (2017). Chemical composition, antibacterial, and antifungal activities of the essential oil of Thymus numidicus Poiret from Algeria. Phytothérapie, 2017, 1–6. ‏

Merabti, B. (2016). Identification, composition et structure des populations de culicidiens dans la région de Biskra, effet des facteurs écologiques sur la distribution saisonnière, essais de contrôle [Identification, composition and structure of culicidian populations in the Biskra region, effect of ecological factors on seasonal distribution, control trials]. University de Mohamed Khider, Biskra (in French).

Merabti, B., Boumaza, M., Ouakid, M. L., Carvajal, T. M., & Harbach, R. E. (2021). An updated checklist of the mosquitoes (Diptera: Culicidae) present in Algeria, with assessments of doubtful records and problematic species. Zootaxa, 5027(4), 515–545. ‏

Mockute, D., & Bernotiene, G. (2004). 1,8-Cineole-caryophyllene oxide chemotype of essential oil of Thymus serpyllum L. growing wild in Vilnius (Lithuania). Journal of Essential Oil Research, 16(3), 236–238. ‏

Mohammadi, A., Sani, T. A., Ameri, A. A., Imani, M., Golmakani, E., & Kamali, H. (2015). Seasonal variation in the chemical composition, antioxidant activity, and total phenolic content of Artemisia absinthium essential oils. Pharmacognosy Research, 7(4), 329–334.

Mohammed, H. A. (2022). Phytochemical analysis, antioxidant potential, and cytotoxicity evaluation of traditionally used Artemisia absinthium L. (wormwood) growing in the central region of Saudi Arabia. Plants, 11(8), 1028.

Moussa, M. B., Belhadi, A., Douak, I., Laouar, A. K., Boudjemaa, S., Hadef, Y., & Bouaricha, A. (2020). Composition chimique et activité antibactérienne de l'huile essentielle de Thymus algeriensis Boiss & Reut. de la région de Batna Algérie [Chemical composition and antibacterial activity of the essential oil of Thymus algeriensis Boiss & Reut. from the Batna region of Algeria]. Revue Aurassienne du Laboratoire, 851(3), 85–90 (in French).

Msaada, K., Salem, N., Bachrouch, O., Bousselmi, S., Tammar, S., Alfaify, A., ... & Marzouk, B. (2015). Chemical composition and antioxidant and antimicrobial activities of wormwood (Artemisia absinthium L.) essential oils and phenolics. Journal of Chemistry, 2015(1), 804658.

Nabti, I., & Bounechada, M. (2019). Larvicidal activities of essential oils extracted from five Algerian medicinal plants against Culiseta longiareolata Macquart. larvae (Diptera: Culicidae). European Journal of Biology, 78(2), 133–138.‏

Nguyen, H. T., & Németh, Z. É. (2016). Sources of variability of wormwood (Artemisia absinthium L.) essential oil. Journal of Applied Research on Medicinal and Aromatic Plants, 3(4), 143–150.

Nguyen, H. T., Radácsi, P., Gosztola, B., Rajhárt, P., & Németh, É. Z. (2018). Accumulation and composition of essential oil due to plant development and organs in wormwood (Artemisia absinthium L.). Industrial Crops and Products, 123, 232–237. ‏

Nieto, G. (2017). Biological activities of three essential oils of the Lamiaceae family. Medicines, 4(3), 63.‏

Nikolić, B., Matović, M., Mladenović, K., Todosijević, M., Stanković, J., Đorđević, I., ... & Tešević, V. (2019). Volatiles of Thymus serpyllum obtained by three different methods. Natural Product Communications, 14(6), 1934578X19856254.

Nikolić, M., Glamočlija, J., Ferreira, I. C., Calhelha, R. C., Fernandes, Â., Marković, T., ... & Soković, M. (2014). Chemical composition, antimicrobial, antioxidant and antitumor activity of Thymus serpyllum L., Thymus algeriensis Boiss. and Reut and Thymus vulgaris L. essential oils. Industrial Crops and Products, 52, 183–190.‏

Nutrizio, M., Pataro, G., Carullo, D., Carpentieri, S., Mazza, L., Ferrari, G., ... & Režek Jambrak, A. (2020). High voltage electrical discharges as an alternative extraction process of phenolic and volatile compounds from wild thyme (Thymus serpyllum L.): in silico and experimental approaches for solubility assessment. Molecules, 25(18), 4131. ‏

Orav, A., Raal, A., Arak, E., Müürisepp, M., & Kailas, T. (2006). Composition of the essential oil of Artemisia absinthium L. of different geographical origin. Chemistry, 55(3), 155–165.

Petrović, S. S., Ristić, M. S., Petrović, N. V., Lazić, M. L., Francišković, M., & Petrović, S. (2014). Chemical composition and antioxidative activity of essential oil of Thymus serpyllum L. Hemijska Industrija, 68(3), 389–397.

Pohlit, A. M., Lopes, N. P., Gama, R. A., Tadei, W. P., & de Andrade Neto, V. F. (2011). Patent literature on mosquito repellent inventions which contain plant essential oils – a review. Planta Medica, 77(6), 598–617.‏

Porto, C. D., & Decorti, D. (2012). Analysis of the volatile compounds of aerial parts and essential oil from Thymus serpyllum L. cultivated in North East Italy by HS-SPME/GC-MS and evaluation of its flavouring effect on ricotta cheese. Journal of Essential Oil Bearing Plants, 15(4), 561–571. ‏

Preljević, K., Pašić, I., Vlaović, M., Matić, I. Z., Krivokapić, S., Petrović, N., ... & Perović, S. (2024). Comparative analysis of chemical profiles, antioxidant, antibacterial, and anticancer effects of essential oils of two Thymus species from Montenegro. Fitoterapia, 174, 105871.

Pruteanu, A., Popescu, C., Vladut, V., & Gageanu, G. (2018). Biochemical analysis of some vegetal extracts obtained from indigenous spontaneous species of Thymus serpyllum L. Romanian Biotechnological Letters, 23(5), 14013–14024. ‏

Quezel, P., & Santa, S. (1962–1963). Nouvelle flore de l'Algérie et des régions désertiques méridionales [New flora of Algeria and the southern desert regions]. Editions du Centre National de la Recherche Scientifique,‏ Paris (in Freinch).

Rezaeinodehi, A., & Khangholi, S. (2008). Chemical composition of the essential oil of Artemisia absinthium growing wild in Iran. Pakistan Journal of Biological Sciences, 11(6), 946–949. ‏

Riahi, L., Ghazghazi, H., Ayari, B., Aouadhi, C., Klay, I., Chograni, H., ... & Zoghlami, N. (2015). Effect of environmental conditions on chemical polymorphism and biological activities among Artemisia absinthium L. essential oil provenances grown in Tunisia. Industrial Crops and Products, 66, 96–102.‏

Roselló, J., Llorens-Molina, J. A., Larran, S., Sempere-Ferre, F., & Santamarina, M. P. (2024). Biofilm containing the Thymus serpyllum essential oil for rice and cherry tomato conservation. Frontiers in Plant Science, 15, 1362569.

Salaria, D., Rolta, R., Patel, C. N., Dev, K., Sourirajan, A., & Kumar, V. (2022). In vitro and in silico analysis of Thymus serpyllum essential oil as bioactivity enhancer of antibacterial and antifungal agents. Journal of Biomolecular Structure and Dynamics, 40(20), 10383–10402. ‏

Santana-Rios, G., Orner, G. A., Amantana, A., Provost, C., Wu, S. Y., & Dashwood, R. H. (2001). Potent antimutagenic activity of white tea in comparison with green tea in the Salmonella assay. Mutation Research/Genetic Toxicology and Environmental Mutagenesis, 495(1–2), 61–74. ‏

‏Sefidkon, F., Dabiri, M., & Mirmostafa, S. A. (2004). The composition of Thymus serpyllum L. oil. Journal of Essential Oil Research, 16(3), 184–185.‏

Shafique, S., Shafique, S., & Sahar, S. (2020). Thymus serpyllum – a novel biocontrol agent against leaf spot pathogens of Solanum melongena. Pakistan Journal of Weed Science Research, 27(2), 469–479.

Sharopov, F. S., Sulaimonova, V. A., & Setzer, W. N. (2012). Composition of the essential oil of Artemisia absinthium from Tajikistan. Records of Natural Products, 6(2), 127–134. ‏

Silver, J. B. (2008). Mosquito ecology: Field sampling methods. 3rd edition. New-York.

Šojić, B., Tomović, V., Kocić-Tanackov, S., Kovačević, D. B., Putnik, P., Mrkonjić, Ž., ... & Pavlić, B. (2020). Supercritical extracts of wild thyme (Thymus serpyllum L.) by-product as natural antioxidants in ground pork patties. LWT, 130, 109661.

Soltanbeiglu, S., Bavani, M. M., Vahedi, M., & Chavshin, A. R. (2022). Susceptibility of Culiseta longiareolata to insecticides in West Azerbaijan province, Northwestern Iran. Veterinary Research Forum, 13(3), 435–437.

Taherkhani, M., Rustaiyan, A., Rasooli, I., & Taherkhani, T. (2013). Chemical composition, antimicrobial activity, antioxidant and total phenolic content within the leaves essential oil of Artemisia absinthium L. growing wild in Iran. African Journal of Pharmacy and Pharmacology, 7(2), 30–36.

Tamma, N., Benchikha, N., Messaoudi, M., Caruso, G., Emran, T. B., Atoki, A. V., & Adeniyi, A. I. (2024). Chemical composition and biological properties of Thymus capitatus plants from Algerian high plains: A comparative and analytical study. Open Chemistry, 22(1), 20230192.

Tariku, Y., Hymete, A., Hailu, A., & Rohloff, J. (2011). In vitro evaluation of antileishmanial activity and toxicity of essential oils of Artemisia absinthium and Echinops kebericho. Chemistry and Biodiversity, 8(4), 614–623. ‏

Tazabayeva, K. A., & Sylibaeva, B. (2018). Chemical composition of the essential oil and flavonoids of Thymus serpyllum L., growing on territory of the East Kazakhstan. Acta Poloniae Pharmaceutica – Drug Research, 75(6), 1329–1337.

Trendafilova, A., Todorova, M., Ivanova, V., Zhelev, P., & Aneva, I. (2023). Essential oil composition of ten species from sect. serpyllum of genus Thymus growing in Bulgaria. Diversity, 15(6), 759.‏

Uniyal, A., Tikar, S. N., Singh, R., Shukla, S. V., Agrawal, O. P., Veer, V., & Sukumaran, D. (2014). Repellent effect, knockdown study and electrophysiological responses of essential oils against Aedes aegypti. Journal of Entomology and Zoology Studies, 2, 351–357. ‏

Vassiliou, E., Awoleye, O., Davis, A., & Mishra, S. (2023). Anti-inflammatory and antimicrobial properties of thyme oil and its main constituents. International Journal of Molecular Sciences, 24(8), 6936.

Verma, R. S., Padalia, R. C., Saikia, D., Chauhan, A., Krishna, V., & Sundaresan, V. (2016). Chemical composition and antimicrobial activity of the essential oils isolated from the herbage and aqueous distillates of two Thymus species. Journal of Essential Oil Bearing Plants, 19(4), 936–943.

Wesołowska, A., Grzeszczuk, M., Jadczak, D., Nawrotek, P., & Struk, M. (2015). Comparison of the chemical composition and antimicrobial activity of Thymus serpyllum essential oils. Notulae Botanicae Horti Agrobotanici Cluj-Napoca, 43(2), 432–438.‏

Wichtl, M., & Anton, R. (1999). Plantes thérapeutiques: Tradition, pratique, officinale [Therapeutic plants: Tradition, practice, officinal]. Science et Thérapeutique, Paris (in French).

Wojtunik‐Kulesza, K. A., & Wiśniewska, R. (2022). Interactions of selected monoterpenes with iron and copper ions based on Ferrozine and CUPRAC methods – the preliminary studies. Chemistry and Biodiversity, 19(8), e202200461.

Xie, Y., Jin, H., Yang, X., Gu, Q., & Zhang, D. (2020). Toxicity of the essential oil from Thymus serpyllum and thymol to larvae and pupae of the housefly Musca domestica L. (Diptera: Muscidae). Environmental Science and Pollution Research, 27, 35330–35340. ‏

Zahed, K., Souttou, K., Hamza, F., & Zamoum, M. (2021). Chemical composition and larvicidal activities in vitro and in vivo of essential oils of Thymus vulgaris (L) and Lavandula angustifolia (Mill) against pine processionary moth Thaumetopoea pityocampa Den. & Schiff. in Ain Defla (Algeria). Journal of Plant Diseases and Protection, 128(1), 121–137.