Effect of formulated essential oil of Ajwain, Trachyspermum ammi (L.) on some biological and physiological parameters of cotton bollworm, Helicoverpa armigera

Document Type : Complete paper

Authors

1 Department of Plant Protection, Faculty of Agriculture, Azarbaijan Shahid Madani university-Tabriz-Iran

2 Department of Plant Protection, Faculty of Agriculture, Azarbaijan Shahid Madani University, Tabriz, Iran

3 Department of Plant Protection, Faculty of Agriculture, Ege University, 35100 Izmir, Türkiye

Abstract

Unrestricted use of chemical insecticides, especially at high concentrations, has reduced their efficacy. Plant-based pesticides, due to their relatively high safety levels to humans and environment as well as effective control on insects, have become a suitable alternative to conventional pesticides. The aim of this study was to investigate the biological and physiological effects of the formulated essential oil of Trachyspermum ammi (L.) on the larvae of Helicoverpa armigera Hüb. The obtained essential oil was formulated into an emulsion using Tween 20 as emulsifier. Chemical analysis of the extracted essential oil indicated that Terpinen (14.4%), Cymene (18.5%), and Thymol (61.6%) were the major compounds. The essential oil of T. ammi have a significant repellency rate on third and sixth instar larvae of cotton bollworm at different observed time intervals, with an average repellency rate of 61-76%. Regarding fumigant toxicity, the highest concentration of the essential oil (500 µL/L) resulted in 100% larval mortality. The LC50 value for fumigant toxicity measured as 82.73 µL/L. The duration of larval development significantly increased in treatments compared to control. In EO concentrations of 2.5-1.25 percent an increase was observed respectively in the duration of the larval stage by 9 and 7 days. The pupae and adult’s emergence affected in dose dependent manner. At concentration of 50 µl/l, 89% inhibition of protease enzyme recorded. With regards to the biological and physiological effects observed in the formulated T. ammi essential oil, with complimentary researches, it could be used in integrated management programs of cotton bollworm.

Keywords

Main Subjects


Extended Abstract

Introduction

The use of chemical pesticides as the most common method of pest control with less selective properties, has resulted in problems such as residual, development of resistance in pests, negative effects on natural enemies, and consequently pest outbreaks. These problems have led to develop alternative control methods with fewer adverse effects in recent years. Plant extracts and essential oils with acceptable insecticidal, antifeedant, repellent, attractant, lower cost and less environmental pollution properties can be used in pest management programs.

Cotton bollworm is one of the most important pests of agricultural crops, vegetables and even ornamentals. It is a polyphagous insect and the use of various pesticides in high concentrations has led to develop resistance to a wide range of chemical pesticides from different groups. Alternative methods for controlling this pest include agricultural methods, biological control, the use of resistant plants, the use of plant metabolites, and integrated pest management recommended by researchers in the field of plant conservation.

Trachyspermum ammi is an annual plant belonging to the apiaceae family which has a high percentage of essential oil in its seeds. Numerous studies have been conducted on the biological effects of this plant essential oil on insects, including the fumigant effect, repellency, digestive system disruption, physiology and other sublethal effects on biology of insects. In recent years, with the expansion of attention to biological pesticides, the insecticidal effects of this plant have also been well studied, including its fumigant, lethal effects, antifeedant, larvicidal, and repellent effects against various pests.

The present study aimed to investigate the various biological, behavioral, and physiological effects of T. ammi essential oil emulsion on cotton bollworm.

 

Material and Methods

Insects reared in controlled condition on artificial diet. The essential oil of T. ammi seeds was prepared by water distillation using Clevenger apparatus. To prepare the emulsion of EO, Tween 20 used as emulsifier.

Gas chromatography mass spectrometry (GC-MS) was used to identify the essential oil chemical composition.

Biological effects including ovicidal, repellent activity, fumigant toxicity and sublethal effects including larval life span, adults and pupa emergence, mortality during generation recorded. Inhibitory activity of essential oils in different concentrations on alpha-amylase, protease and acetylcholine esterase, carried out using standard protocols.

The data was analyzed using SPSS v: 26 software. In case of observing mortality in control treatments, the Abbot formula was used to correct the data. SPSS software and F-test were used for data analysis and mean comparisons.

 

Results and discussion

The main identified compounds of the T. ammi essential oil extract, using gas chromatography, include thymol, cymene, and terpinene, with percentages of 61.6%, 18.5%, and 14.4%, respectively.

The ovicidal assays indicate that, with increasing the concentration of the EO hatching rate decreased. the concentration of 2.5 percent showed up to 80% ovicidal toxicity. The LC50 value obtained was 0.632 percent. The slope of the concentration-mortality line was calculated to be 1.36.

The highest concentration of the oil, 500 microliters per liter, caused 100% mortality of larvae, and after that, with a concentration of 250 µl/l air, 81.66% mortality recorded. The LC50 and LC70 values were calculated as 82.73 and 151.14 µl/l air, respectively. The slope of the concentration-mortality line was also calculated as 2.002.

About repellent activity of T. ammi essential oil, a repellency rate of 61 to 76 percent was observed for 3rd and 64 to 76 percent for 6th larvae instars at different time intervals of 6 to 16 hours.

with increasing the concentrations, the length of larval developmental period increased, and a direct relationship between the concentration of the essential oil and the length of the larvae developmental duration was recorded. At concentrations of 2.5 and 1.25 percent of T. ammi essential oil, the length of the larvae developmental period increased to 9 and 7 days, respectively, in comparing with the control.

About sublethal effects, the highest concentration of essential oil (5%) completely prevented the pupation and adult’s emergence. Even at the lowest concentration of T. ammi essential oils (0.039%), only 28% pupation and adult emergence appeared, indicating the significant effect of the essential oil on the population of cotton bollworm. Also some deformities were also observed in the emerged pupas.

The essential oil of T. ammi caused significant inhibition of alpha-amylase, protease and acetylcholinesterase enzymes activity in 6th instar larvae of cotton bollworm.

 

Conclusion

According to the results of current research, the formulated essential oil of T. ammi has good potential to manage cotton bollworm. The observed biological and physiological effects indicate the significant impact of this essential oil in reducing the population of cotton bollworm.

Aazza, S., Lyoussi, B., & Miguel, MG. (2011). Antioxidant and antiacetylcholinesterase activities of some commercial essential oils and their major compounds. Molecules, 16, 7672–7690. 
Abd-El-Aziz, SE. (2011). Control strategies of stored product pests. Journal of Entomology, 8, 101-122.
Abdelgaleil, SAM., & El-Sabrout, AM. (2018). Anti-nutritional, antifeedant, growth-disrupting and insecticidal effects of four plant essential oils on Spodoptera littoralis (Lepidoptera: Noctuidae). Journal of. Crop Protection, 7 (2), 135-150
Adams, RP., & Sparkman, OD. (2007). Review of identification of essential oil components by Gas Chromatography /Mass Spectrometry. Journal of the American Society for Mass Spectrometry, 18, 803-806.
Aggarwal, N., Brar, DS. & Basedow, T. (2006). Insecticide resistance management of Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) and its effect on pests and yield of cotton in North India. Journal of Plant Diseases and Protection, 113 (3), 120–127.
Bisrat, D. & Jung, C. (2020). Insecticidal toxicities of three main constituents derived from Trachyspermum ammi (L.) sprague ex turrill fruits against the small Hive beetles, Aethina tumida Murray. Molecules, 25, 1100; doi:10.3390/molecules25051100.
Chaubey, MK. (2018). Study of insecticidal properties of Trachyspermum ammi and Mentha arvensis essential oils against Sitophilus zeamais L.(Coleoptera: Curculionidae). Current Life Sciences, 4(1), 10-17.
Conti, B., Canale, A., Cioni, PL., & Flamini, G., (2010). Repellence of essential oils from tropical and Mediterranean lamiaceae against Sitophilus zeamais. Bulletin of Insectology, 63, 197–202.
Darrag, HM., Mohammed Alhajhoj, R. & Ezzat Khalil, H., (2021). Bio-insecticide of Thymus vulgaris and Ocimum basilicum extract from cell suspensions and their inhibitory effect against Serine, Cysteine, and Metalloproteinase of the Red palm weevil (Rhynchophorus ferrugineus). Insects, 2021, 12, 405. https:// doi.org/10.3390/insects12050405.
Darvishzadeh, A., Hosseininaveh, V. & Salimian Rizi, S. (2014). Enzymatic activity of α-amylase in alimentary tract Spodoptera littoralis (Boisduval) (Lepidoptera: Noctuidae): Characterization and Compartmentalization. Arthropods, 3(3), 138-146
El-Sabrout, A., Zahran, HE-D. & Abdelgaleil, S. (2018). Effects of essential oils on growth, feeding and food utilization of Spodoptera littoralis larvae. Journal of Entomology, 15(1),36–46.
Fernández-Peña, L., Gutiérrez-Muro, S., Guzmán, E., Lucia, A., Ortega, F., & Rubio R. (2019). Oil-In-water microemulsions for thymol solubilization. Colloids and Interfaces, 3: 64. https://doi.org/10.3390/colloids3040064
Fite, T., Tadele T., Mulugeta, N., Tebekew, D., & Waktole, G. (2018). Management of Helicoverpa armigera (Lepidoptera: Noctuidae) by nutritional indices study and botanical extracts of Millettia ferruginea and Azadirachta indica. Advances in Entomology, 6: 235-255. doi:10.4236/ae.2018.64019
Gandomi, H., Abbaszadeh, S.,   JebelliJavan, A. & Sharifzadeh, A. (2013). Chemical constituents, antimicrobial and antioxidative effects of Trachyspermum ammi essential oil. Journal of Food Processing and Preservation, 38, 1690–1695.
Habashi, AS., Safaralizadeh, MH. & Safavi, SA. (2011). Fumigant toxicity of Carum copticum L. oil against Tribolium confusum du Val, Rhyzopertha dominica F. and Oryzaphilus surinamensis L. Munis Entomology and Zoology, 6(1), 282-289.
Hyder, M., Li, Y., Wang, M., Mao, J., Mari, J.M., Bukero, A., Soomro, H.U., Bukero, A.A. & Zhang, L. (2022). Insecticidal activity, chemical constituents of Trachyspermum ammi, withania coagulans and Murraya koenigii ethanloic extracts against Bemisia tabaci. Brazilian Journal of Biology, 84, e260298 https://doi.org/10.1590/1519-6984.260298
Isman, MB. (2000). Plant essential oils for pest and disease management. Crop Protection, 19, 603-608.
Isman, MB. (2006). Botanical insecticides, deterrents, and repellents in modern agriculture and an increasingly regulated world. Annual Review of Entomology, 51, 45–66.
Jansen, JP., Lauvaux, S., Gruntowy, J. & Denayer, J., (2017). Possible synergistic effects of fungicide-insecticide mixtures on beneficial arthropods. International Organization for Biological and Integrated Control, 125, 28-35.
Jeyasankar, A., Chennaiyan, V. & Chinnamani, T. (2016). Evaluation of five essential plant oils as a source of repellent and larvicidal activities against larvae of Tribolium castaneum (Herbst) (Coleoptera: tenebrionidae). Journal of Entomology, 13, 98-103.
Jukic, M., Politeo, O., Maksimovic, M., Milos, M. & Milos, M. (2007). In vitro acetylcholinesterase inhibitory properties of thymol, carvacrol and their derivatives thymoquinone and thymohydroquinone. Phytotherapy Research, 21(3), 259-261. doi: 10.1002/ptr.2063. PMID: 17186491.
Kedia, A., Prakash, B., Mishra, PK., Dwivedy, AK. & Dubey, NK. (2015). Trachyspermum ammi L. essential oil as plant based preservative in food system, Industrial Crops and Products, 69,104-109.
Khajeh, M., Yamini,. Y, Sefidkon, F. & Bahramifar, N. (2004). Comparison of essential oil composition of Carum copticum obtained by supercritical carbon dioxide extraction and hydrodistillation methods. Food chemistry, 86(4), 587-591.
Kim, SI., Ahn, YJ. & Kwon, HW. (2012). Toxicity of aromatic plants and their constituents against coleopteran stored products insect pests. New perspectives in plant protection, 93-120.
Koul, O., Walia, S. & Dhaliwal, GS. (2008). Essential oils as green pesticides: Potential and constraints. Biopesticides International, 4, 63–84.
Kumar Chaubey, M. (2018). Study of insecticidal properties of Trachyspermum ammi and Mentha arvensis essential oils against Sitophilus zeamais L. (Coleoptera: Curculionidae). Current Life Sciences, 4 (1), 10-17.
Kumar, V. & Kumar, P. (2019). Pesticides in agriculture and environment: impacts on human health. In: Contaminants in agriculture and environment: health risks and remediation. Haridwar, India: Agro Environ Media Agriculture and Environmental Science Academy.  pp 76–95.
Lee, SC., Seo, SM., Huh, MJ., Kwon, JH., Nam, I., Park, JH. & Park, IK. (2020). Behavioral and electrophysiological effects of ajowan (Trachyspermum ammi Sprague) (Apiales: apiaceae) essential oil and its constituents on nymphal and adult bean bugs, Riptortus clavatus (Thunberg) (Hemiptera: alydidae). Insects, 11(2):104. doi: 10.3390/insects11020104. PMID: 32033226; PMCID: PMC7074463.
Liao, M., Xiao, J-J., Zhou, L-J., Yao, X., Tang, F., Hua, R-M., Wu, X-W. & Cao, H-Q. (2017). Chemical composition, insecticidal and biochemical effects of Melaleuca alternifolia essential oil on the Helicoverpa armigera. Journal of Applied Èntomology, 141, 721-728.
Liao, M., Xiao, JJ., Zhou, LJ., Yao, X., Tang, F., Hua, RM., Wu, XW. & Cao, HQ. (2017). Chemical composition, insecticidal and biochemical effects of Melaleuca alternifolia essential oil on the Helicoverpa armigera. Journal of Applied Entomology, 141,721-728.
Martin, TO., Chou, OG., Hala N’klo, F., Vassal, JM. & Vaissayre, M. (2000). Pyrethroid resistance in the cotton bollworm, Helicoverpa armigera (Hubner), in West Africa. Pest Management Science, 56, 549-554.
Mervat, AK., Ahmed, AF. & Moustafa, HZ. (2012). Toxicological and biochemical studies of lufenuron, chlorfluazuron and chromafenozide against Pectinophora gossypiella (Saunders). Egyptian Academic Journal of Biological Sciences, 4, 37- 47.
Miyazawa, M. & Yamafuji, C. (2005). Inhibition of acetylcholinesterase activity by bicyclic monoterpenoids. Journal of Agricultural and Food Chemistry, 53, 1765-1768.
Moein, MR., Zomorodian, K., Pakshir, K., Yavari, F., Motamedi, M. & Zarshenas, MM. (2015). Trachyspermum ammi (L.) Sprague: chemical composition of essential oil and antimicrobial activities of respective fractions. Journal of Evidence-Based Complementary and Alternative Medicine, 20, 50–56.
Mohagheghzadeh, A., Faridi,. P. & Ghasemi, Y. (2007). Carum copticum Benth. Hook., essential oil chemotypes. Food Chemistry, 100(3), 1217-1219.
Mojarab-Mahboubkar, M., Jalali Sendi, J. & Aliakbar, A. (2015). Effect of Artemisia annua L. essential oil on toxicity, enzyme activities, and energy reserves of cotton bollworm Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae). Journal of Plant Protection Research, 55 (4), 371-377 (In Persian).
Nenaah, GE. (2014). Bioactivity of powders and essential oils of three asteraceae plants as post -harvest grain protectants against three major coleopteran pests. Journal of Asia -Pacific Entomology, 17, 701 -709.
Nerio, LS., Olivero-Verbel, J. & Stashenko, E. (2010). Repellent activity of essential oils: A review of Bioresource Technology, 101, 372–378.
Pandey, SK/, Upadhyay, S. & Tripathi, AK. (2009). Insecticidal and repellent activities of thymol from the essential oil of Trachyspermum ammi (Linn) Sprague seeds against Anopheles stephensi. Parasitology research, 105(2), 507-512.
Piri, A., Sahebzadeh, N., Zibaee, A., Sendi, JJ., Shamakhi, L. & Shahriari, M. (2020). Toxicity and physiological effects of ajwain (Carum copticum, Apiaceae) essential oil and its major constituents against Tuta absoluta (Meyrick)(Lepidoptera: Gelechiidae). Chemosphere, 256, 127103.
Rahali, N., Mehdi, S., Younsi, F., Boussaid, M. & Messaoud, C. (2017). Antioxidant, α-amylase, and acetylcholinesterase inhibitory activities of Hertia cheirifolia essential oils: Influence of plant organs and seasonal variation, International Journal of Food Properties, 20,1637-1651, DOI: 10.1080/10942912.2017.1352597.
Regnault-Roger, C., Vincent, C. & Arnason, JT. (2012). Essential oils in insect control: Low-risk products in a high-stakes world. Annual Review of Entomology, 57, 405–424.
Regnault-Roger, C. (1997). The potential of botanical essential oils for insect pest control. Integrated Pest Management Reviews, 2(1), 25-34.
Saad, NY. & Muller, CD. (2013). Major bioactivities and mechanism of action of essential oils and their components, Flavour and Fragrance Journal, 28(5), 269–279.
Said-Al-Ahl, H., Hikal, WM. & Tkachenko, KG. (2017). Essential oils with potential as insecticidal agents: A review. International Journal of Environmental Plant Management, 3, 23–33.
Saroukolai, AT., Nouri-Ganbalani, G., Hadian, J. & Rafiee-Dastjerdi, H. (2014). Antifeedant activity and toxicity of some plant essential oils to Colorado potato beetle, Leptinotarsa decemlineata Say (Coleoptera: Chrysomelidae). Plant Protection Science, 50, 207–216.
Shorey, HH. & Hale RL. (1965). Mass rearing larvae of nine Noctuid species on a simple artificial medium. Journal of Economic Entomology, 58, 522-524.
Siriporn, P. & Mayura, S. (2012). The effects of herbal essential oils on the oviposition-deterrent and ovicidal activities of Aedes aegypti (Linn.), Anopheles dirus (Peyton and Harrison) and Culex quinquefasciatus (Say). Trop Biomed, 29(1), 138-50.
Soni, R., Sharma, G. & Dut Jasuja, N. (2016). Essential oil yield pattern and antibacterial and insecticidal activities of Trachyspermum ammi and Myristica fragrans. Scientifica, https://doi.org/10.1155/2016/1428194.
Thomas, M., (1999). Ecological approaches and the development of “truly integrated” pest management. Proceedings of the National Academy of Sciences, 96, 5944 -5951.
Torabi-Pour, H., Shayeghi, M., Vatandoost, HM. & Abai M. (2017). Larvicidal effects and phytochemical evaluation of essential oils of Trachyspermum ammi and Ziziphora clinopodioides against larvae Anopheles stephensi. Journal of Herbmed Pharmacology, 6,185-190.
Torres-Vila, LM., Rodriguez-Molina, MC., Lacasa-Plasencia, A., Bielza-Lino, P. & Rodriguez-del-Rincon, A. (2002). Pyrethroid resistance of Helicoverpa armigera in Spain: Current status and agroecological perspective, Agriculture, Ecosystems and Environment, 93, 55-66.
Tripathi, AK., Upadhyay, S., Bhuiyan, M. & Bhattacharya, PR. (2009). A review on prospects of essential oils as biopesticides in insect-pest management. Journal of Pharmacognosy Phytotherapy, 1, 52–63.
Upendhar, S., Sree, KV., Satyanarayana, J. & Singh, TVK. (2017). Cypermethrin and methomyl resistance in Helicoverpa armigera (Hübner). International Journal of Economic Plants, 4, 70–75.
Vitali, L., Beghelli, D., Biapa, P., Bistoni, O., Cappellacci, L., Damiano, S., Lupidi, G., Maggi, F., Orsomando, G., Papa, F., Petrelli, D., Petrelli, R., Quassinti, L., Sorci, L., Zadeh, M. & Bramucci, M., (2016). Diverse biological effects of the essential oil from Iranian Trachyspermum ammi, Arabian Journal of Chemistry, 9(6), 775-786.
Warikoo, R., Wahab, N. & Kumar, S. (2011). Oviposition-altering and ovicidal potentials of five essential oils against female adults of the dengue vector, Aedes aegypti L. Parasitology Research, 109(4), 1125-31. doi: 10.1007/s00436-011-2355-y. Epub 2011 Mar 29. PMID: 21445613.
Yang, Y., Isman, MB. & Tak, JH. (2020). Insecticidal activity of 28 essential oils and a commercial product containing Cinnamomum cassia bark essential oil against Sitophilus zeamais Motschulsky. Insects, 11, 1-15.
Yazdani, E., Sendi, JJ. & Hajizadeh, J. (2014). Effect of Thymus vulgaris L. and Origanum vulgare L. essential oils on toxicity, food consumption, and biochemical properties of lesser mulberry pyralid Glyphodes pyloalis Walker (Lepidoptera: Pyralidae). Journal of Plant Protection Research, 54, 53-61.
Yeom, HJ., Kang, JS., Kim, GK. & Park, IK. (2012). Insecticidal and acetylcholine esterase Inhibition activity of apiaceae plant essential oils and their constituents against adults of german cockroach (Blattella germanica). Journal of Agricultural and Food Chemistry, 60 (29), 7194-7203.
Younsi, F., Trimech, R., Boulila, A., Ezzine, O., Dhahri, S., Boussaid, M. & Messaoud, C. (2016). Essential oil and phenolic compounds of Artemisia herbaalba (Asso.): Composition, antioxidant, antiacetylcholinesterase and antibacterial activities. International Journal of Food Properties, 19, 1425–1438.
Youssefi, MR., Tabari, MA., Esfandiari, A., Kazemi, S., Moghadamnia, AA., Su, TS., Dall'Acqua, S., Benelli, G. & Maggi, F. (2019). Efficacy of two monoterpenoids, carvacrol and thymol, and their combinations against eggs and larvae of the West Nile vector Culex pipiens. Molecules, 5;24(10): 1867. doi: 10.3390/molecules24101867. PMID: 31096594; PMCID: PMC6572342.