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The present work was to evaluated the insecticidal activity of the essential oil extracted from an aromatic plant Artemisia herba alba (Lamiaceae) on larvaes and insects of Tribolium confusum. The results obtained showed that the essential oil A. herba alba revealed significant antiradical activities against the DPPH radical and a powerful reducing power. The repellent effect and the toxicity by contact were tested at different concentrations (2, 4, 6, 8 μl/ml). The results revealed that the essential oil of A.herba alba was moderately toxic after 24 h towards the insects and the larvaes. While the repellent effect showed that Artemisia oil had a very strong repellent effect on the larvae’s. These results added to those obtained by other authors on the usefulness and effectiveness of essential oils in the control of different.
Laboratory of Biotechnology and Biomonitoring of the Environment and Oasis Ecosystems (LBBEEO), Faculty of Sciences of Gafsa, University of Gafsa, Tunisia
Dorsaf Ben Jeddou
Laboratory of Biotechnology and Biomonitoring of the Environment and Oasis Ecosystems (LBBEEO), Faculty of Sciences of Gafsa, University of Gafsa, Tunisia
Hadil Kadri
Laboratory of Biotechnology and Biomonitoring of the Environment and Oasis Ecosystems (LBBEEO), Faculty of Sciences of Gafsa, University of Gafsa, Tunisia
Guesmi Fatma
Laboratory of Biotechnology and Biomonitoring of the Environment and Oasis Ecosystems (LBBEEO), Faculty of Sciences of Gafsa, University of Gafsa, Tunisia
Abassi Raoudha
Laboratory of Biotechnology and Biomonitoring of the Environment and Oasis Ecosystems (LBBEEO), Faculty of Sciences of Gafsa, University of Gafsa, Tunisia
Najla Hfaiedh
Laboratory of Biotechnology and Biomonitoring of the Environment and Oasis Ecosystems (LBBEEO), Faculty of Sciences of Gafsa, University of Gafsa, Tunisia
*Address all correspondence to: bouzenna.hafsia@hotmail.fr
1. Introduction
Plants represent an immense source of complex chemical molecules exploited by man in the perfume, food, cosmetics and pharmaceutical industries. Most plants contain essential oils; they are then called “aromatic plants”. These essential oils were found in many parts of the plant: wood, leaves, fruits, bark, seeds and roots. These are complex mixtures made up of several dozen or even more than a hundred compounds, mainly terpenes and aromatic compounds.
In addition, plant production of economic interest often suffered damages caused by pests, disease agents such as viruses, bacteria, insects or fungi. To control the crop enemies, producers had often used synthetic chemicals, including insecticides, fungicides, bactericides and herbicides. Unfortunately, the application of these chemicals had caused unexpected problems both in terms of soil contamination by non-degradable chemical molecules and on human and animal health by the harmful residues that persist on food.
In order to limit these drawbacks, several academic and public laboratories had taken an interest in the development of biological control [1]. In this work, was interested in a medicinal plant, it is Artemisia herba-alba, for which, it’s essential oils was extracted, and evaluated its possible interest in the context of biological control.
A. herba-alba has been used in traditional medicine by many cultures since ancient times. It is a very popular remedy that is often used to facilitate digestion, calm abdominal pain and certain liver ailments. Its roots were indicated against certain nervous disorders [2]. Their aqueous extract were traditionally used in Jordan as an antidote against the venoms of several types of snakes and scorpions. In North Africa, it was known to treat bronchitis, abscess, diarrhea and as a vermifuge. Its oil essential had many activities including antioxidant, hypoglycemic, antifungal, antibacterial [3, 4].
In this context, the study was evaluated the bioinsecticide effect of the essential oil of A. herba-alba on adults and larvae of T. confusum.
2.1 Extraction of essential oil from A. herba alba
2.1.1 Plant material
The biomass used for the extraction of essential oils was composed of the aerial parts of A. herba alba. They were collected from Jbel Mdhila mountain of the Gafsa region in April 2022. The aerial parts of this plant were dried in the open air, away from light, for 8 days. After drying, the dried plant material was ground in a mortar to increase the solvent-sample contact surface.
2.1.2 Chemicals
Acetone, DPPH, BHT, potassium ferricyanide, trichloroacetic acid were purchased from Sigma-Aldrich Chemicals Co. (St. Louis, MO, USA).
2.2 Extraction of volatile oils
The extraction of essential oils was done by the method of hydrodistillation: Hydro-distillation is a technique for extracting essential oils that is done using a simple distillation. In a 1 liter flask, 500 mL of distilled water was added with a few pumice stones to regulate the heating and about 50 g of the dried leaves of plant. The water was brought to a boil, taking care not to heat until completely dry. Thus, the water vapor entrains the volatile organic substances which condense in the refrigerator then flow drop by drop and are collected at the other end of the assembly in the graduated cylinder. The volume of the distillate is approximately 130 mL.
2.3 Determination of extraction yield
The yield of essential oils is defined as being the ratio between the mass of the essential oil obtained and the mass of the plant material treated.
R=(mm0)×100
With R: essential oil yield (%), m: mass in grams of the essential oil, m0: mass in grams of dry plant matter.
2.4 Study of biological activities
2.4.1 Antioxidant activity of essential oil.
The antioxidant activity was tested using two methods: scavenging of the free radical DPPH and determination of its reducing power (FRAP).
2.4.2 The DPPH• (2,2-diphenyl-1-picrylhydrazyl) test
DPPH• (2,2-Diphenyl-1-picrylhydrazyl) is a stable purplish-colored free radical that absorbs at 517 nm. In the presence of anti-radical compounds, the DPPH• radical is reduced and changes color by turning yellow. This method is based on measuring the capacity of antioxidants to scavenge the 2,2-diphenyl-1-picrylhydrazil (DPPH•) radical. The latter is reduced to the form of hydrazine (non-radical) by accepting a hydrogen atom. The effect of oils on DPPH is measured by the procedure described by Öztürk et al. [5]. The percentage of DPPH anti-radical activity is calculated according to the equation:
2.4.3 Reducing power (FRAP)
The reducing power of ferric ions (Fe3+) of an extract was evaluated using the method described by Oyaizu [6]. A volume of 1.25 mL of a phosphate buffer solution (0.1 M, pH 6.6) and 1.25 mL of potassium ferricyanide (1% w/v) were mixed with 500 μL of an extract solution at concentrations variables. The reaction mixture was incubated for 30 minutes at 50°C, then a volume of 1.25 mL of 10% w/v trichloroacetic acid was added to stop the reaction, the mixture was centrifuged at 1500 × g for 10 min. A volume of 1.25 mL of the supernatant is mixed with 1.25 mL of distilled water and 250 μL of a 0.1% w/v solution of FeCl3 then incubated for 10 minutes. The absorbance of the mixture was measured at 700 nm. The greater the absorbance of the reaction mixture, the greater the reducing power. BHT was used as a positive control with the same concentrations (50–300 μg/mL) and under the same experimental conditions was used as a standard antioxidant and the results are presented as the change in absorbance with concentration. From the graph the effective concentration (EC50) is determined which gives an absorbance of 0.5. The lower the EC50, the stronger the reducing power. The test measuring the reducing power was repeated in three trials for each extract.
2.5 Study of the insecticidal and larvicidal activity of the essential oil of A. herba alba
The toxicity tests of A. herba-alba on insects and larvae of T. confusum were carried out according to two modes of penetration, one penetration by contact and the other by repellent effect.
2.6 Evaluation of the toxicity of this oil by contact effect
After preparing the doses, each solution was spread evenly on a disc of Wattman-type filter paper previously placed in Petri dishes of the same diameter. After evaporation of the dilution solvent, all the boxes are infested by 10 insects or larvae of T. confusum. We carried out three repetitions for the four doses of essential oil tested, and the same for the control not treated with the essential oil. A count of the dead insects is carried out after 24 hours of this treatment.
2.7 Test to evaluate the repellent effect of essential oil on insects and larvaes
The repellent effect of essential oils against adults and larvae of T. confusum was evaluated using the technique described by McDonald et al. [7] using the preferential area method on filter paper. In this experiment, the 9 cm diameter filter paper discs used for this purpose were divided into two equal parts. Four different essential oil contents were used in this test: 2, 4, 6, 8 μl/ml [8]. 0.5 mL of each dose was placed on one half of the disc, while the other half received only acetone. The two discs were placed in the open air for 10 minutes, in order to evaporate the acetone. After acetone drying. The filter paper disc was placed in a 9 cm diameter Petri dish. A batch of 10 individuals of T. confusum were placed in the center of filter paper. The boxes are placed under ambient temperature conditions to calculate the percentage of repellent value for each essential oil. The percentage of repulsion is thus calculated according to the following formula:
Percent Repulsion(PR%)=((NC−NT)/NC)×100
Where: NC: number of the individual present on the part of the disc treated only with acetone NT: number of the individual present on the part of the disc treated with the prepared dose. The average percentage repellency for each oil is calculated (PR) and assigned to one of the different repellent classes varying from 0 to V [7] which showed in Table 1.
Class
Repulsion interval
Property of the treated substance
Class 0
PR ≤ 0,1%
Not repellent
Class I
0,1 < PR ≤ 20%
Very weakly
Class II
20 < PR ≤ 40%
Repellent weakly
Class III
40 < PR ≤ 60%
Repellent moderately
Class IV
60 < PR ≤ 80%
Repellent
Classe V
80 < PR ≤ 100%
Very repellent
Table 1.
Percentage of repulsion (PR) according to the classification of Donald et al. [7].
2.8 Data analysis method
2.8.1 Mortality correction by Abbott’s method
The effectiveness of this essential oil is evaluated by mortality. The results of the tests carried out do not only represent the mortality caused by the oil but there is also the natural mortality. This mortality is corrected using the Abbott formula.
MC(%)=((MT−Mt)/(100−Mt))×100
_ MC %: percentage of corrected mortality.
_ Mt.: percentage of mortality obtained in the control population.
_ MT: percentage of mortality obtained in the treated population.
The extraction by hydrodistillation of the aerial parts of A. herba-alba, obtained the essential oil of yellow color and very strong smell. The yield of the essential oil obtained in relation to the total weight of the dry plant is 0.16%. The yield was low compared to that obtained by the same plant in different regions of Algeria (from 0.2 to 0.95%) [2, 9]. That of Spain was 0.41 to 2.30% [10] and that of Jordan 1.3% [11]. This difference in yields can be attributed not only to the geographical origin of the plant, but also to many factors such as: growth stage, pedoclimatic conditions, place of production, etc. Several studies had shown the influence of the vegetative cycle and the extraction technique on the yield and quality of EO.
3.2 Evaluation of antioxidant activity
In order to evaluate the antioxidant activity of A. herba alba essential oil, different antioxidant tests (DPPH and FRAP) were used.
3.2.1 Anti-radical activity against DPPH radical
The antioxidant effect was assessed by the DPPH test as the ability of DPPH •, compounds with antiradical activity, to scavenge free radicals. From these results, it deduced that the evolution of the anti-free radical activity is studied in vitro against the radical DPPH is dose-dependent (Figure 1). Indeed, the increase in the concentration of the oil is followed by an increase in the anti-DPPH activity. These results were also expressed by IC50. Indeed, the IC50 parameter is defined as being the concentration of the substrate which leads to a loss of 50% of the activity. Indeed, the oil with the lowest IC50 value exerts the most powerful antiradical activity. Indeed, the oil reported a value of IC50 = 160 μg/mL. This reflected a powerful antioxidant power by comparing it with the standard (ascorbic acid) which has the IC50 value = 210 μg/mL.
Figure 1.
Antiradical activity of the essential oil of Artemisia herba alba. Each value represents the mean ± SD (n = 3 trials for each sample). HE: Essential oil, Ac: ascorbic acid.
3.2.2 Iron reduction test (FRAP)
The reducing power was based on the reduction of the ferric ion (Fe3+) to ferrous ion (Fe2+) by transfer of an electron or donation of a hydrogen atom. The Figure 2 represented the reducing power of essential oil of A. herba alba and the BHT standard. The results showed that the reducing power increased with the concentration. The absorbance at 700 nm of the oil increased proportionally to its concentration. However, in comparison with BHT (used as a control), the reducing power of this oil was lower. Indeed, even at high concentration (300 μg/mL), we observed an absorbance of 0.32 for oil against 1.00 for BHT.
Figure 2.
Graphic representation of the reducing power of A. herba alba and BHT each value represents the mean ± SD (n = 3 trials for each sample).
3.2.3 Insecticidal and larvicidal activity of the essential oil of A. herba alba
To evaluate the insecticidal and larvicidal effect of the essential oil of A. herba alba the mortality rate of adults and larvae’s by contact and repellent method were estimated.
3.2.3.1 Assessment of adult and larval mortality by contact effect
The corrected mortalities were mentioned in Figures 3 and 4. The results indicated that the larvae were more resistant than the insects. The results showed an increase in the concentration-corrected mortality rate. As an aromatic plant, the volatility of the essential oil due to the montepenes and the major Camphor compound of this oil gives it an effectiveness in controlling stock insect pests. These results confirm other inferences reporting Chaied et al. [12] that this oil showed good insecticidal activity.
Figure 3.
Corrected mortality percentage of T. confusum insects after 24 h of exposure to A.herba alba essential oil. Each value represents the mean ± SD (n = 3 replicates with 10 insects each).
Figure 4.
Percentage of corrected mortality of larvae of T. confusum after 24 h of exposure to A.herba alba essential oil. Each value represents the mean ± SD (n == 3 replicates with 10 insects each).
3.2.3.2 Determination of the repellent activity of the essential oil
The results of the evaluation of the repellent effects of essential oil of A. herba alba on the larvae and adults of T. confusum were presented in Tables 2 and 3. The percentage of repellent essential oil used increases according to the dose and time. Our deductions revealed that the highest dose of A. herba alba had a strong repellent effect of 100% on T. confusum larvaes and adults respectively. From these results, it was concluded that higher concentration of essential oil resulted in maximum pest repellency compared to lower concentrations. According to the classification established by Mc. Donald et al. [7], it can be concluded that the oil recorded a very strong repellent effect against adults and larvae of T. confusum. Many researchers have pointed out that certain plant essential oils exhibited strong repellent effects against harmful storage [13, 14, 15].
30 min
60 min
90 min
120 min
Repellent effects
Classe
Repellent effects
Classe
Repellent effects
Classe
Repellent effects
Classe
2
13.3 ± 8.08
Class I
20 ± 6
Class I
46.66 ± 3.05
Class III
53.33 ± 3.05
Class II
4
20 ± 4
Class I
36.03 ± 3.05
Class II
42.2 ± 3.4
Class III
60 ± 4
Class III
6
46.66 ± 4.6
Class III
62.5 ± 3.03
Class IV
73.33 ± 4.6
Class IV
100 ± 00
Classe V
8
53.33 ± 1.1
Class III
64.4 ± 3.35
Class IV
73.33 ± 4.6
Class IV
100 ± 00
Classe V
Table 2.
Repellent effects on T. confusum larvae exposed to A. herba alba essential oil.
30 min
60 min
90 min
120 min
Effet répulsif
Classe
Effet répulsif
Classe
Effet répulsif
Classe
Effet répulsif
Classe
2
6
Class I
80 ± 1.15
Class IV
80 ± 3.4
Class IV
77.77 ± 3.84
Class IV
4
80 ± 1.15
Classe V
80 ± 1.15
Classe V
80 ± 2
Classe V
100 ± 00
Classe V
6
80 ± 2
Classe V
93.33 ± 1.15
Classe V
93.33 ± 1.15
Classe V
100 ± 00
Classe V
8
86.66 ± 2.3
Classe V
100 ± 00
Classe V
100 ± 00
Classe V
100 ± 00
Classe V
Table 3.
Repellent effects on adults of Tribolium confusum exposed to the essential oil of A. herba alba.
The use of bio-insecticides was a major and effective potential for crop protection. The use of essential oils extracted from aromatic plants has demonstrated their insecticidal activity against insect pests of stocks. In this work, initially, the evaluation of the antioxidant activity by the DPPH free radical scavenging method and the FRAP test showed that the essential oil has a significant antioxidant power. On the other hand, treatment with A. herba alba essential oil on T. confusum larvae and insects gave a repellent effect and a toxic effect. At the end of this study, the results obtained are in agreement with previous results. They largely confirm the usefulness of essential oils in the pest control program, particularly in P. interpunctella and E. kuheniella and in different orders of insects.
This research was supported and funded by the Ministry of Higher Education and Scientific Research (MHESR, Tunisia).
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Written By
Bouzenna Hafsia, Dorsaf Ben Jeddou, Hadil Kadri, Guesmi Fatma, Abassi Raoudha and Najla Hfaiedh
Submitted: 24 January 2023Reviewed: 20 February 2023Published: 16 June 2023
Open access peer-reviewed chapter
