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Chemical Compositions and Fumigation Effects of Essential Oils Derived from Cardamom, Elettaria cardamomum (L.) Maton, and Galangal, Alpinia galanga (L.) Willd Against Red Flour Beetle, Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae)

A peer-reviewed version of this preprint was published in:
Plants 2024, 13(13), 1845. https://doi.org/10.3390/plants13131845

Submitted:

02 May 2024

Posted:

07 May 2024

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Abstract
The prevention of destruction caused by red flour beetle, Tribolium castaneum (Herbst), using essential oils derived from certain herbal plants presents an interesting alternative to synthetic insecticides, known for their harm to both consumers and the environment. The research aimed to investigate the chemical compositions and fumigating effects of essential oils derived from cardamom (Elettaria cardamomum (L.) Maton) and galangal (Alpinia galanga (L.) Willd) against T. castaneum. Chemical compositions were analyzed using GC-MS, while fumigation bioassay was conducted via a vapor-phase test. Experimental setup followed factorial design in CRD with 5 replications. Factor A encompassed 4 types of essential oils: the manually extracted essential oils from cardamom leaves (MCL) and galangal leaves (MGL), the commercially produced essential oils from cardamom seeds (CCS) and galangal rhizomes (CGR). Factor B consisted of 7 concentrations: 0, 50, 100, 150, 200, 250, and 300 µL/L air. Results indicated that MCL contained eucalyptol, trans-calamenene, and isospathulenol as its main compounds. Similarly, CCS was characterized by camphene and eucalyptol. MGL exhibited caryophyllene, aciphyllene, and α-bisanolene. Lastly, CGR was composed of methyl cis-cinnamate, safrole, and p-vinylphenyl isothiocyanate. Study noted that 250 µL/L air over 168 hours of CGR, demonstrated 94% fumigation efficiency against adult T. castaneum. These results suggest the potential of galangal rhizome derived essential oil as an insecticide for managing T. castaneum populations in agricultural product storage.
Keywords: 
Zingiberaceae
;  
essential oil
;  
stored insect pest
;  
plant secondary metabolite
;  
GC-MS
Subject: 
Biology and Life Sciences  -   Insect Science

1. Introduction

Stored-product insects threaten global grain stores, causing losses ranging from 10% in temperate regions to nearly 50% in humid tropical areas [1]. Grains and flour, vital sources of carbohydrates worldwide, are particularly prone to infestation, leading to reduced quantity and quality. Post-harvest storage, a critical phase in the agricultural supply chain, directly impacts stored commodities' quality and quantity, but it is often disrupted by pest infestations. These pests exploit favorable conditions within warehouses, evading detection and causing both quality degradation and quantitative losses, commonly known as shrinkage [2]. Various insect species attack grain stores, causing significant losses estimated between 5% to 30% of total global agricultural production [3]. Notably, the red flour beetle poses a serious threat by contaminating grain with shed skins, feces, and dust, affecting cereal flour and stored foods. Postharvest losses due to insect pests can reach 30% to 40%, posing a significant threat to human and livestock health [4].
The red flour beetle, Tribolium castaneum (Herbst, 1797), is a highly damaging storage pest, threatening stored products and the food economy. Despite its classification as a secondary pest, T. castaneum infests damaged grains during harvesting, affecting various stored food items, leading to significant economic losses [5]. This infestation causes agglomeration, discoloration, and spoilage, with profound economic implications [6]. It is recognized as one of the most destructive cosmopolitan pests of stored products due to its broad host range and rapid infestation capabilities. Additionally, T. castaneum infestations raise temperature and humidity within storage environments, promoting bacterial and fungal growth [7]. Exposure to T. castaneum may trigger allergic responses and contribute to product deterioration [8]. The development of resistance to various insecticides, including malathion and phosphine, poses a significant challenge for T. castaneum control, increasing the risk of food contamination. With its ability to breed year-round in warm climates, T. castaneum poses a continuous threat to global food security, especially in tropical and subtropical regions [9]. Effective monitoring and control measures are essential to mitigate its impact on the cereal value chain and ensure food safety.
The extensive use of chemical insecticides to safeguard stored products from infestation has resulted in insect resistance, leading to environmental pollution and harm to non-target organisms [10]. While phosphine, malathion, and deltamethrin effectively control pests like T. castaneum, their excessive application has caused resistance, prompting the need for alternative approaches [11,12,13]. Insects pose significant threats to global food security, necessitating safer alternatives due to resistance and adverse effects associated with current insecticides [10]. Despite phosphine's widespread use as a fumigant, resistance in T. castaneum has emerged, indicating the need for safer options. Dependence on chemical pesticides has caused harm to non-target organisms and environmental pollution [14]. Integrated pest control strategies, which integrate plant essential oils with chemical pesticides, offer a promising approach to reduce environmental impact and ensure sustainable pest management.
Plant secondary metabolites, including essential oils, play a crucial role in plant-insect interactions, enhancing plant resistance to insects. These oils possess various pest control properties such as larvicidal, repellent, and antifeedant effects, making them valuable for integrated pest management [11,15]. They degrade into non-toxic compounds, minimizing risks to non-target organisms and humans, highlighting the importance of natural insecticides for food safety. Essential oils from aromatic plants effectively combat insect pests through fumigation, contact, repellency, and antifeedant actions [16], primarily due to their volatile compounds, particularly monoterpenes and sesquiterpenes, which act as fast-acting neurotoxins in insects, disrupting various biological processes, including growth and reproduction [17]. Concerns about the drawbacks of synthetic insecticides, such as resistance and environmental pollution, have driven interest in botanical insecticides, especially essential oils [10], offering specificity, biodegradability, and low mammalian toxicity, thus promising for integrated pest management. Their natural composition and low persistence mitigate the risk of resistance development [18].
Elettaria cardamomum (L.) Maton, commonly referred to as cardamom, is an aromatic herb from the Zingiberaceae family, globally recognized for its commercial significance, earning it the title "Queen of spices." While native to India and Sri Lanka, cardamom cultivation has expanded to diverse tropical and subtropical regions worldwide [19]. The unique aroma and bioactive compounds of cardamom contribute to its pharmaceutical and nutraceutical properties, rendering it a valuable botanical resource [20,21]. It finds applications across various sectors, including food, perfumery, and traditional medicine, with traditional uses spanning stimulating, stomachic, diuretic, carminative, and anti-infective properties [22]. Renowned for its diverse biological effects, cardamom exhibits therapeutic properties such as antispasmodic, stimulant, anthelmintic, antimicrobial, antiviral, antioxidant anticancer, anti-inflammatory, and insecticidal effects [23,24,25,26]. The essential oil of cardamom, primarily comprising 1,8-cineole and α-terpinyl acetate, demonstrates toxic effects on various stored-product pests, including coleopteran and lepidopteran insects, through contact and fumigant actions [27].
Alpinia galanga (L.) Willd., commonly known as galangal, is an aromatic herb belonging to the Zingiberaceae family. It has a rich history of diverse applications, including alleviating stomachache, displaying antibacterial and antifungal properties, exhibiting antitumor effects, aiding in antiulcer activity, showing antiallergic effects, possessing antioxidant properties, antiplasmid, antimicrobial activities, and demonstrating insecticidal activity [28,29,30,31]. Extensively cultivated in Southeast Asian countries such as the Philippines, Indonesia, Thailand, India, and China, it holds significance in various traditional medical systems, including Thai, Ayurveda, Unani, and Chinese folk medicine [32]. It is traditionally considered an important source for medication, culinary products and cosmetics in Asia. For medical purposes, galangal has been widely using as encompass anti-inflammatory, antipyretic, emmenagogue, carminative, abortifacient, and aphrodisiac properties [33]. Moreover, studies have investigated the toxicity of essential oil extracted from A. galanga rhizomes against Bactrocera dorsalis (Hendel), providing insights into its potential as a natural insecticide [31]. Therefore, the aim of this study was to assess the chemical compositions and fumigating effects of essential oils derived from cardamom (Elettaria cardamomum (L.) Maton) and galangal (Alpinia galanga (L.) Willd) against adults of the red flour beetle, Tribolium castaneum (Herbst). Additionally, gas chromatography-mass spectrometry (GC-MS) analysis was conducted to determine their chemical composition.

2. Results

2.1. Chemical Composition

The manually extracted essential oil from cardamom leaves (MCL) contained a total of 28 chemical components, accounting for 90.77 %. The primary compounds were eucalyptol (25.20%), trans-calamenene (13.49%) and isospathulenol (13.09%). Additionally, it included ylangenal (4.96%), α-guaiene (4.13%), ledol (4.10%), carveol (3.18%), α-cubebene (3.13%), junenol (2.71%), isoaromadendrene epoxide (1.93%), epicubenol (1.63%), trans-verbenol (1.42%) and caryophyllene oxide (1.36%) (Table 1). The commercially produced essential oil from cardamom seeds (CCS) comprised a total of 34 constituents, accounting for 98.09%. The main compounds include camphene (46.06%) and eucalyptol (31.21%), along with linalyl acetate (4.72%), α-pinene (4.69%), linalool (2.49%), terpineol (1.93%) and nerolidol (1.06%) (Table 2).
Besides cardamom, eucalyptol, also known as 1,8-cineole, is also found in other plants such as eucalyptus (Eucalyptus globulus Labill.) and rosemary (Rosmarinus officinalis L.). Ashokkumar et al. [24] note that all cardamom accessions contain 1,8-cineole as the second most significant monoterpene constituent, ranging from 15.2% to 49.4%. Nonetheless, certain studies have noted lower proportions of α-terpinyl acetate and linalool, accompanied by higher percentages of 1,8-cineole in cardamom essential oil originating from various countries, with 1,8-cineole ranging from 25.6% to 26.71% [34,35]. Similarly, Noshad and Behbahani [36] identified 23 compounds in E. cardamomum essential oil, with eucaluptol being the major compound at 31.51%. Alanazi et al. [37] reported that GC/MS revealed 19 chemical constituents, indicating 98.2% of E. cardamomum essential oil, with the most compounds were monoterpenes constituents, such as 1,8-cineole (34.3%), α-terpinyl acetate (23.3%), and α-pinene (17.7%). The variation in the chemical composition of E. cardamomum essential oil may be attributed to factors such as chemotypes, geographical locations, season at the time of plant collection, stage of plant development, culture climate, extraction techniques, plant varieties, and plant part used, which can affect subsequent biological activities [38,39,40] observed fluctuations in volatile constituents of cardamom essential oil grown in distinct zones of Idukki hills, with changes in α-terpinyl acetate and 1,8-cineole levels across different zones as the season progressed.
The manually extracted essential oil from galangal leaves (MGL) consisted of 25 components, accounting for 95.74%. The predominant compounds were caryophyllene (24.17%), aciphyllene (18.31%) and α-bisanolene (10.69%). Additionally, it contained ç-elemene (7.67%), α-farnesene (6.98%), farnesyl butanoate (6.08%), humulene (4.52%), caryophyllene oxide (4.15%), n-hexadecanoic acid (2.30%), cedrene (1.89%), 11,11-dimethyl-4,8-dimethylenebicyclo[7.2.0]undecan-3-ol (1.57%) and ç-himachalene (1.37%) (Table 3). The commercially produced essential oil from galangal rhizomes (CGR) included 27 compositions, accounting for 98.73%. The key compounds were methyl cis-cinnamate (47.28%), safrole (19.82%) and p-vinylphenyl isothiocyanate (11.52%). It also contained eucalyptol (7.65%), α-pinene (5.96%), α-phellandrene (1.46%) and terpineol (1.12%) (Table 4).
The chemical composition analysis of galangal leaves, as reported by Menon [41], revealed β-caryophyllene (40.5%) and fenchyl acetate (20.7%) as the main compounds. Conversely, cubenol (28.4%), carotol (26.7%), and α-fenchyl acetate (30.5%) were predominant in stem, rhizome, and root. In northern India, A. galanga from the Himalayan region showed 1,8-cineole (39.4% and 32.5%) and β-pinene (11.9% and 22.7%) as primary compounds in its rhizome and leaf oils, respectively, with camphor (12.8%) also present in the leaf oil [42,43]. 1,8-cineole serves as the marker compound for Alpinia spp., with eucalyptol identified as the major constituent in A. galanga, as reported by Singh et al. [44]. Additionally, Abeywickrama et al. [45] confirmed 1,8-cineole as the major constituent in the essential oil of A. calcarata. In the oil from A. galanga in Tenom, Sabah, Malaysia, Nampoothiri et al. [46] identified major components in A. galanga, including 1,8-cineole, α-terpineol, and germacrene-D. Wu et al. [47] identified 51 components in the essential oil extracted from A. galanga rhizomes, with eucalyptol and pinenes being prominent. Various studies have investigated the chemistry of galangal rhizome, including analyses conducted by De Pooter et al. [48]. These findings highlight the need for further research on plant cultivation and essential oil standardization to account for genetic, geographic, physiological, and harvest timing factors that contribute to variations in essential oil composition within the same plant species [49].
By comparing the chemical composition of the essential oils from cardamom, E. cardamomun, and galangal, A. galanga, it can be seen that the essential oils from MCL and CCS contained eucalyptol as a component and importantly, they were the same (Table 5). Plant extracts and essential oils commonly contain insecticidal monoterpenoids like limonene, linalool, and terpineol, carvacrol, which are effective against stored product insects [50,51]. These natural products exhibit insecticidal and acaricidal effects by disrupting biological pathways and inducing tissue injuries that may lead to the generation of free radicals [52]. Several monoterpenoids have been identified for their fumigant toxicity against various insects, including α-terpineol against T. confusum [53], β-pinene against Sitophilus oryzae [54], and 1,8-cineol against both S. oryzae and T. confusum [55,56]. Additionally, α-pinene was found toxic to S. oryzae [55], while p-cymene, α-terpinene, α-terpineol, and terpinene-4-ol showed potential fumigant toxicity to S. oryzae as well [50]. Moreover, γ-terpinene and terpinene-4-ol were reported as promising fumigants against T. confusum and Ephestia kuehnielle [57]. Additionally, α-pinene and β-caryophyllene were found to have repellent action against S. zeamais [58,59], while α-pinene also exhibited potential fumigant activity against S. zeamais [60].
Acetylcholinesterase (AChE) is crucial in insect and tick nervous systems, breaking down acetylcholine to transmit neuronal signals [61]. Plant-derived volatiles, such as those affecting the octopaminergic mode of action, have neurotoxic effects [62]. While some studies suggest monoterpenoids may inhibit AChE activity [63], others argue that terpenoid toxicity doesn't always correlate with AChE inhibition [54]. The mode of action of monoterpenes can vary, potentially acting on the octopaminergic nervous system or inhibiting cytochrome P450-dependent mono-oxygenases [64]. Auti and Kulkarni [65] found significant AChE inhibition with E. cardamomum essential oil, suggesting neuroprotective effects [66]. Terpenoid compounds like 1,8-cineole, α-pinene, β-pinene, and carvone are identified as primary agents inhibiting AChE activity in essential oils [65]. Additionally, camphene attracts both sexes of Rhynchophorus ferrugineus adults, leading to mortality by blocking their air holes, causing asphyxiation [67]. Essential oils affect insect respiratory, nervous, and hormonal systems, often leading to death [68]. Linalool inhibits AChE and interacts with receptors, showing insecticidal activity [69]. Various compounds like menthol, methonene, limonene, and linalool are toxic to S. oryzae by inhibiting acetylcholine esterase enzyme [54]. Moreover, 1,8-cineole inhibits AChE activity in T. castaneum larvae and has insecticidal properties against post-harvest pests [56]. Linalool affects insect nervous systems by influencing ion transport and acetylcholine esterase release [70]. Caryophyllene oxide's fumigant toxicity may result from inhibiting the mitochondrial electron transport system, affecting insect respiration [71]. Lastly, menthyl cinnamate's moderate activity stems from its double bond and benzene ring configuration [72]. It's worth noting that the activity and mode of action of essential oil compounds can differ from those of individual components [73].

2.1. Chemical Composition

The types of essential oil had a significantly different (P < 0.01) effect on the mortality of adult T. castaneum at 24, 48, 72, 96, 120, 144, and 168 hours. CGR resulted in the highest mortality of adult T. castaneum in all tested periods, with 7.14±5.76%, 11.14±7.90%, 17.14±11.01%, 22.57±15.98%, 36.00±23.30%, 53.71±31.80%, and 64.57. ±37.54 %, respectively. Significant differences were also observed when compared with MCL, CCS and MGL (Table 6). The concentrations of essential oils exhibited a statistically significant difference (P < 0.05) in their effect on the mortality of adult T. castaneum at 48, 72, 96, 120, 144, and 168 hours. A concentration of 300 µL/L air showed the highest mortality 10.50±7.72%, 13.50±9.00%, 21.50±11.82%, 32.00±19.39%, 45.50±27.29%, and 54.00±33.92 %, respectively. However, no statistical difference was found when compared with a concentration of 250 µL/L air. Additionally, a significantly difference (P < 0.01) was observed at 24 hours, with the highest mortality at a concentration of 300 µL/L air being 6.50±3.00%, but no statistical difference was found when compared with a concentration of 250 µL/L air (Table 6).
The interaction between types of essential oils and concentrations was found to have no effect on the mortality of adult T. castaneum within 24-72 hours after the fumigation bioassay. However, significant differences (P < 0.05) were observed in the mortality of adult T. castaneum at 96 hours due to the interaction between essential oil types and concentrations. CGR at the concentrations of 150 and 250 µL/L air resulted in the highest mortality of 38%, but no difference was observed when compared to a concentration of 300 µL/L air, nor was there a significant difference when compared to CCS at the concentrations of 250 and 300 µL/L air. Furthermore, the effect of interaction between essential oil types and concentrations on the mortality of adult T. castaneum was found to be significantly different (P < 0.01) at 120, 144, and 168 hours. CGR at a concentration of 150 µL/L air exhibited the highest mortality at 120 hours, with 64.00±33.62%, but no difference was observed compared to the concentrations of 250 and 300 µL/L air, which resulted in mortalities of 56.00±13.42% and 54.00±33.62%, respectively. At 144 hours, CGR at the concentrations of 250 and 300 µL/L air had the highest mortality of adult T. castaneum, with 80.00±7.07%, and 80.00±22.36%, respectively, with no significant difference observed compared to a concentration of 150 µL/L air, where the mortality was 78.00±23.87%. Finally, at 168 hours, CGR at a concentration of 300 µL/L air resulted in the highest mortality of adult T. castaneum, at 96.00±5.48%. However, no difference was observed compared to the concentrations of 150, 200, and 250 µL/L air, which had the mortality 84.00±15.17%, 84.00±11.40%, and 94.00±5.48%ม respectively (Table 6).
Wang et al. [74] support the notion that the fumigant effect of essential oils was closely linked to dosage and exposure duration. Result indicated that the essential oil of CGR at 250-300 µL/L air gave the highest mortality (94-96%) of adult T. castaneum. Our results concurred with Wu et al. [47] demonstrated the potent fumigant activity of A. galanga rhizome essential oil against Lasioderma serricorne, surpassing other essential oils studied previously like those from Pistacia lentiscus L., Elsholtzia stauntonii Benth., and Agastache foeniculum (Pursh) Kuntze in fumigant toxicity against cigarette beetles [75,76,77]. This enhanced efficacy may be attributed to key compounds like methyl cis-cinnamate found in A. galanga rhizomes, which are widely used in pharmaceuticals, cosmetics, and the food industry and have documented repellent, insecticidal, and larvicidal properties [78,79,80,81].
Specifically, methyl cinnamate has shown insecticidal effects against S. oryzae and Musca domestica adults, while ethyl cinnamate exhibits antifeedant effects against Spodoptera littoralis and Hylobius abietis, and propyl cinnamate displayed insecticidal effects on M. domestica adults [79,82,83,84]. Fujiwara et al. [85] highlighted the superior larvicidal activity of methyl cinnamate against Ae. aegypti, suggesting the potential of cinnamates as alternatives to conventional insecticides. Moreover, A. galanga rhizome essential oil contains primary constituents like 1,8-cineol and α-terpineol. Several other studies have identified 1,8-cineole as effective insecticidal and oviposition repellents across various insect species [86,87,88]. Research on essential oils from diverse plants has revealed the deterrent effects of 1,8-cineole on neonate larvae of the codling moth and its efficacy in repelling or controlling adult stages of urban insect pests [72,89].

3. Materials and Methods

3.1. Insect Raring

The red flour beetles, Tribolium castaneum (Herbst), were sourced from rice bran commonly utilized as animal feed, specifically collected for experimental purposes. Adult males and females were grouped into 30 pairs. They were fed a mixture of coarsely ground wheat flour mixed and rice bran in a ratio of 7:10, housed within 4 L plastic container, and securely sealed. Ventilation for the containers was scheduled every 3 days. They were maintained within a growth chamber at a 26 ± 5 °C and a relative humidity of 75 ±5%. The 14-day-old adult red flour beetle will be utilized in all bioassays.

3.2. Preparation of Essential Oils

The leaves of cardamom (Elettaria cardamomum (L.) Maton) and galangal (Alpinia galanga (L.) Willd) were collected in November 2022 in Maha Sarakham, Thailand (16°10’38” N 103°18’03” E). Plant materials were washed, roughly chopped, and dried in hot air oven at 65 °C for 2 days, after which they were stored in a plastic bag. Following this, 100 g of dried plant parts were placed in a 2000 mL flask, and 1000 mL of distilled water was added. The mixture underwent water distillation at 100 ± 20 °C for 4 hours using an essential oil extractor. Subsequently, the resulting essential oil underwent purification by centrifuging at 10000 rpm for 10 minutes to separate any remaining water from the essential oil. The pure essential oils were then transferred into sealed amber glass vials and stored in a refrigerator at 4 °C until they were ready for further use in bioassays.
The 100% pure essential oils extracted from the seeds of cardamom, E. cardamomum, and the rhizome of galangal, A. galanga, were obtained commercially from the Aroma & More Shop, 333/19, Moo 19, Bangbuathong-Suphanburi Road, Laharn Sub-district, Bangbuathong District, Nonthaburi Province, 11110, Thailand.

3.3. Chemical Compositon Analysis

The essential oils of cardamom and galangal, both the manually extracted and commercially produced, were analyzed for their chemical composition using the method outlined by Satongrod et al. [90], employing a gas chromatograph-mass spectrometer model Clarus 680 (PerkinElmer USA). The column utilized was an Rtx-5MS capillary type, with a length of 30 m, diameter of 0.32 mm, and thickness of 1 µm. An essential oil concentration of 100000 ppm, with a volume of 1 µL, was injected in split mode (split ratio, 1:100 v/v). Helium gas served as the carrier gas at a flow rate of 1 mL/minutes. The injector temperature was set at 280 °C. Under column conditions, an initial temperature of 45 °C was maintained for 5 minutes before increasing at a rate of 10 °C/minutes to 200 °C, where it was held constant for 5 min. For mass spectrometry conditions the electron impact mode was set at 70 eV. Using a quadrupole mass analyzer, the detector temperature was maintained at 250 °C. Spectra were scanned (m/z) from 40 to 1000 amu.
The essential oils identification was made based on their retention indices (RI) determined with reference to homologues series of C5–C36 (n-alkanes), by comparison of their mass spectra with the reports in the literature using NIST and Wiley version libraries [91], ensuring a quality match of over 80%. Chemical composition data were analyzed by reading retention time and % peak area.

3.4. Effects of Fumigation Bioassay

Fumigation toxicity was assessed through a vapor-phase test conducted in a completely randomized design (CRD) with 5 replicates. Factor A comprised 4 types of essential oils: the manually extracted essential oil from cardamom leaves (MCL), the commercially produced essential oil from cardamom seeds (CCS), the manually extracted essential oil from galangal leaves (MGL), and the commercially produced essential oil from galangal rhizomes (CGR). Factor B involved 7 concentrations: 0 (control, consisting of 100% acetone), 50, 100, 150, 200, 250, and 300 µL/L air, prepared by diluting essential oils with acetone solvent. The test was conducted in closed 40 mL glass vials. One hundred µL of the test solution was applied onto a 2 cm diameter Whatman No.1 filter paper disc, which was then allowed to dry at room temperature for 2 minutes. The filter paper was attached to the bottom surface of the glass vial cap. Five pairs of adult T. castaneum were introduced into each test vial, which was tightly sealed with a screw cap. These vials were then stored in a growth chamber at 26 ± 5 °C and a relative humidity of 75 ± 5%. The number of T. castaneum death was recorded at 24, 48, 72, 96, 120, 144, and 168 hours. Mortality was calculated using the formula [(NC / NT)] x 100 where NC represented the number of dead T. castaneum and NT denoted the total number of T. castaneum used in the test. If the mortality of T. castaneum in the control fell within the range of 5-20%, the mortality of T. castaneum in each treatment needed adjustment using Abbott's formula [92]. Statistical data was conducted using the F-test by analyzing variance (ANOVA) based on the experimental plan factorial in CRD and mean comparisons were made using the least significant difference method (LSD < 0.05).

4. Conclusions

The potential of plant-derived essential oils in controlling stored product pests is significant due to their selectivity and minimal impact on non-target organisms, with our study emphasizing the notable adulticidal activity of A. galanga rhizome essential oil against T. castaneum, suggesting its promise for pest control in stored products.

Author Contributions

Conceptualization, R.W.; methodology, R.W., P.K. and H.B.; validation, R.W. and P.K.; formal analysis, R.W. and H.B.; data curation, R.W.; writing—original draft preparation, R.W.; writing—review and editing, R.W., P.K. and H.B.; visualization, R.W.; supervision, R.W.; project administration, R.W.; funding acquisition, R.W. All authors have read and agreed to the published version of the manuscript.

Funding

This research project was financially supported by Mahasarakham University, Thailand.

Data Availability Statement

The original contributions presented in the study are included in the article, further inquiries can be directed to the corresponding author.

Acknowledgments

This research project was financially supported by Mahasarakham University, Maha Sarakham, Thailand. The authors would like to acknowledge the Department of Agricultural Technology, Faculty of Technology, Mahasarakham University, Maha Sarakham, Thailand for laboratory support. Laboratory assistance from Mr. Kamonwan Sanghawit and Miss Jiraporn Krasaetep is gratefully acknowledged.

Conflicts of Interest

The authors declare no conflicts of interest.

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Table 1. The chemical compositions of the manually extracted essential oil from cardamom, E. cardamomun, leaves (MCL).
Table 1. The chemical compositions of the manually extracted essential oil from cardamom, E. cardamomun, leaves (MCL).
No. Compound Retention Time % Peak Area
1 eucalyptol 8.071 25.20
2 carveol 13.841 3.18
3 trans-verbenol 15.560 1.42
4 α-guaiene 26.043 4.13
5 α-cubebene 26.381 3.13
6 trans-calamenene 27.587 13.44
7 caryophyllene oxide 29.656 1.36
8 isospathulenol 30.307 13.09
9 ledol 30.747 4.10
10 junenol 30.945 2.71
11 ylangenal 31.783 4.96
12 isoaromadendrene epoxide 31.950 1.93
13 epicubenol 32.058 1.63
Total 80.28
Table 2. The chemical compositions of the commercially produced essential oil from cardamom, E. cardamomun, seeds (CCS).
Table 2. The chemical compositions of the commercially produced essential oil from cardamom, E. cardamomun, seeds (CCS).
No. Compound Retention Time % Peak Area
1 α-pinene 6.012 4.69
2 eucalyptol 8.026 31.21
3 linalool 10.291 2.49
4 terpineol 14.046 1.93
5 linalyl acetate 16.388 4.72
6 camphene 20.907 46.06
7 nerolidol 28.971 1.06
Total 92.16
Table 3. The chemical compositions of the manually extracted essential oil from galangal, A. galanga, leaves (MGL).
Table 3. The chemical compositions of the manually extracted essential oil from galangal, A. galanga, leaves (MGL).
No. Compound Retention Time % Peak Area
1 γ-elemene 22.766 7.67
2 caryophyllene 24.405 24.17
3 humulene 25.185 4.52
4 aciphyllene 27.159 18.31
5 α-farnesene 27.415 6.98
6 α-bisanolene 27.822 10.69
7 cedrene 28.145 1.89
8 caryophyllene oxide 30.217 4.15
9 11,11-dimethyl-4,8-dimethylenebicyclo[7.2.0]undecan-3-ol 32.096 1.57
10 ç-himachalene 32.814 1.37
11 farnesyl butanoate 39.247 6.08
12 n-hexadecanoic acid 44.776 2.30
Total 89.70
Table 4. The chemical compositions of the commercially produced essential oil from galangal, A. galanga, rhizomes (CGR).
Table 4. The chemical compositions of the commercially produced essential oil from galangal, A. galanga, rhizomes (CGR).
No. Compound Retention Time % Peak Area
1 α-pinene 6.200 5.96
2 α-phellandrene 6.973 1.46
3 eucalyptol 7.924 7.65
4 terpineol 14.044 1.12
5 safrole 22.488 19.82
6 p-vinylphenyl isothiocyanate 22.658 11.52
7 methyl cis-cinnamate 23.274 47.28
Total 94.81
Table 5. Compare the chemical compositions of cardamom, E. cardamomun, and galangal, A. galanga, essential oils in both the manually extracted and the commercially produced types.
Table 5. Compare the chemical compositions of cardamom, E. cardamomun, and galangal, A. galanga, essential oils in both the manually extracted and the commercially produced types.
No. Compound MCL CCS MGL CGR
1 eucalyptol 25.20 31.21 - 7.65
2 camphene - 46.06 - 5.96
3 methyl cis-cinnamate - - - 47.28
4 caryophyllene - - 24.17 -
5 safrole - - - 19.82
6 aciphyllene - - 18.31 -
7 trans-calamenene 13.44 - - -
8 isospathulenol 13.09 - - -
9 p-vinylphenyl isothiocyanate - - - 11.52
10 α-bisanolene - - 10.69 -
11 caryophyllene oxide 1.36 - 4.15 -
12 humulene - - 4.52 -
13 terpineol - 1.93 - 1.12
14 linalool - 2.49 - -
15 α-pinene - 1.05 - -
Table 6. Mortality of adult T. castaneum on cardamom, E. cardamomun, and galangal, A. galanga, essential oils in both the manually extracted and the commercially produced types.
Table 6. Mortality of adult T. castaneum on cardamom, E. cardamomun, and galangal, A. galanga, essential oils in both the manually extracted and the commercially produced types.
Treatments Mortality of adult T. castaneum (%)
24 h 48 h 72 h 96 h 120 h 144 h 168 h
Essential oils
MCL 3.14±3.02b 4.00±3.27b 4.86±3.63b 6.86±5.40c 10.57±9.57c 15.43±12.79c 25.71±16.07c
CCS 1.71±1.80 b 3.71±3.55b 8.00±6.73 b 13.14±9.72b 24.86±15.44b 34.57±20.26b 43.43±24.73b
MGL 1.42 ±1.90b 2.86±2.54b 3.71±3.73 b 4.86±4.14c 6.57±3.98c 9.43±5.00c 12.86±5.76d
CGR 7.14±5.76a 11.14±7.90a 17.14±11.01a 22.57±15.98a 36.00±23.30a 53.71±31.80a 64.57±37.54a
F-test ** ** ** ** ** ** **
LSD 2.87 3.94 4.93 5.41 6.78 6.66 6.38
Concentrations
0 µL/L air 0.00±0.00d 0.00±0.00d 0.00±0.00c 0.00±0.00c 0.00±0.00e 0.00±0.00e 0.00±0.00e
50 µL/L air 2.50±1.91bcd 3.00±2.00cd 4.00±2.83bc 4.00±2.83c 9.00±6.83d 12.00±8.64d 18.00±7.12d
100 µL/L air 4.00±1.63abc 7.00±2.58abc 11.00±7.75a 12.50±6.40b 17.50±9.93cd 28.50±21.60c 38.50±23.35c
150 µL/L air 2.50±3.79bcd 5.00±7.57bcd 9.00±9.66ab 14.00±16.73b 25.50±27.87abc 34.00±31.71c 41.50±31.97bc
200 µL/L air 2.00±2.83dc 4.00±2.83bcd 8.00±5.89ab 12.00±5.74b 22.00±13.00bc 35.00±24.49bc 49.00±30.70ab
250 µL/L air 6.00±8.16ab 8.50±7.19ab 13.50±11.36a 19.00±15.87ab 30.50±21.00ab 43.00±30.61ab 55.00±33.32a
300 µL/L air 6.50±3.00a 10.50±7.72a 13.50±9.00a 21.50±11.82a 32.00±19.39a 45.50±27.29a 54.00±33.92a
F-test * ** ** ** ** ** **
LSD 3.80 5.21 6.53 7.16 8.97 8.81 8.44
Essential oils x Concentrations
MCL 0 µL/L air 0.00±0.00 0.00±0.00 0.00±0.00 0.00±0.00f 0.00±0.00i 0.00±0.00i 0.00±0.00i
MCL 50 µL/L air 0.00±0.00 0.00±0.00 0.00±0.00 0.00±0.00f 0.00±0.00i 0.00±0.00i 10.00±12.25i
MCL 100 µL/L air 6.00±5.48 8.00±4.47 8.00±4.47 8.00±4.47def 12.00±8.37ghi 14.00±11.40ghi 32.00±10.95efg
MCL 150 µL/L air 2.00±4.47 4.00±8.94 4.00±8.94 6.00±8.94ef 6.00±8.94hi 14.00±5.48ghi 20.00±12.25gh
MCL 200 µL/L air 2.00±4.47 4.00±5.48 8.00±8.37 12.00±8.37cdef 16.00±5.48fghi 20.00±10.00fgh 32.00±16.43efg
MCL 250 µL/L air 4.00±5.48 4.00±5.48 6.00±5.48 8.00±4.47def 20.00±12.25efgh 26.00±11.40fg 44.00±16.73e
MCL 300 µL/L air 8.00±4.47 8.00±4.47 8.00±4.47 14.00±5.48cdef 20.00±7.07efgh 34.00±15.17ef 42.00±19.24e
CCS 0 µL/L air 0.00±0.00 0.00±0.00 0.00±0.00 0.00±0.00f 0.00±0.00i 0.00±0.00i 0.00±0.00i
CCS 50 µL/L air 4.00±8.94 4.00±8.94 6.00±8.94 6.00±8.94ef 12.00±8.37ghi 16.00±11.40ghi 24.00±23.02fgh
CCS 100 µL/L air 2.00±4.47 4.00±5.48 4.00±5.48 10.00±0.00def 18.00±10.95fgh 34.00±19.49ef 38.00±21.68ef
CCS 150 µL/L air 0.00±0.00 0.00±0.00 6.00±5.48 12.00±10.95cdef 28.00±19.24defgh 36.00±18.17def 48.00±13.04de
CCS 200 µL/L air 0.00±0.00 2.00±4.47 6.00±8.94 12.00±8.37cdef 36.00±15.17de 50.00±12.25cde 64.00±11.40cd
CCS 250 µL/L air 2.00±4.47 10.00±10.00 20.00±7.70 26.00±5.48abc 38.00±13.04bc 54.00±11.40c 66.00±5.48c
CCS 300 µL/L air 4.00±8.94 6.00±13.42 14.00±16.73 26.00±15.17abc 42.00±25.88cd 52.00±27.75cd 64.00±25.10cd
MGL 0 µL/L air 0.00±0.00 0.00±0.00 0.00±0.00 0.00±0.00f 0.00±0.00i 0.00±0.00i 0.00±0.00i
MGL 50 µL/L air 2.00±4.47 4.00±8.94 6.00±13.42 6.00±13.42ef 8.00±13.04hi 12.00±10.95ghi 14.00±11.40hi
MGL 100 µL/L air 4.00±5.48 6.00±8.94 10.00±14.14 10.00±14.14def 10.00±14.14hi 10.00±14.14ghi 14.00±13.42hi
MGL 150 µL/L air 0.00±0.00 0.00±0.00 0.00±0.00 0.00±0.00f 4.00±8.94hi 8.00±8.37hi 14.00±11.40hi
MGL 200 µL/L air 0.00±0.00 2.00±4.47 2.00±4.47 4.00±5.48f 6.00±5.48hi 8.00±8.37hi 16.00±11.40ghi
MGL 250 µL/L air 0.00±0.00 2.00±4.47 2.00±4.47 4.00±8.94f 8.00±8.37hi 12.00±10.95ghi 16.00±15.17ghi
MGL 300 µL/L air 4.00±5.48 6.00±5.48 6.00±5.48 10.00±7.07def 12.00±4.47ghi 16.00±8.94ghi 16.00±8.94ghi
CGR 0 µL/L air 0.00±0.00 0.00±0.00 0.00±0.00 0.00±0.00f 0.00±0.00i 0.00±0.00i 0.00±0.00i
CGR 50 µL/L air 4.00±5.48 4.00±5.48 4.00±5.48 4.00±5.48f 16.00±11.40fghi 20.00±14.14fgh 24.00±15.17fgh
CGR 100 µL/L air 4.00±5.48 10.00±10.00 22.00±19.24 22.00±19.24bcd 32.00±19.24def 56.00±28.81c 70.00±15.81bc
CGR 150 µL/L air 8.00±8.37 16.00±15.17 26.00±20.74 38.00±14.83a 64.00±33.62a 78.00±23.87ab 84.00±15.17ab
CGR 200 µL/L air 6.00±8.94 8.00±8.37 16.00±11.40 18.00±13.04cde 30.00±15.81def 62.00±14.83bc 84.00±11.40ab
CGR 250 µL/L air 18.00±14.83 18.00±14.83 26.00±16.73 38.00±16.43a 56.00±13.42ab 80.00±7.07a 94.00±5.48a
CGR 300 µL/L air 10.00±14.14 22.00±21.68 26.00±24.08 36.00±35.78ab 54.00±33.62abc 80.00±22.36a 96.00±5.48a
F-test ns ns ns * ** ** **
LSD - - - 14.33 17.94 17.63 16.88
* represents a significant difference at P < 0.05, ** represents a significant difference at P < 0.01, Means followed by the same column followed by the same letter are not significantly different (LSD: P > 0.05).
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