Home Life Sciences Toxicity of Calophyllum soulattri, Piper aduncum, Sesamum indicum and their potential mixture for control Spodoptera frugiperda
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Toxicity of Calophyllum soulattri, Piper aduncum, Sesamum indicum and their potential mixture for control Spodoptera frugiperda

  • Neneng Sri Widayani , Danar Dono EMAIL logo , Yusup Hidayat , Safri Ishmayana and Edy Syahputra
Published/Copyright: September 20, 2023
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Open Agriculture
From the journal Open Agriculture Volume 8 Issue 1

Abstract

Spodoptera frugiperda caused severe damage to the maize plant. Botanical insecticides are a choice to control this pest. This study aims to determine the ethanol extract of Calophyllum soulattri stem bark, methanol extract of Piper aduncum fruit, and Sesamum indicum oil, and their potential mixture for controlling S. frugiperda. The bioassays were carried out in laboratory conditions using second instar S. frugiperda larvae from mass rearing in the laboratory. A toxicity test was performed using the leaf-residual feeding method. The result showed that the mortality of S. frugiperda for C. soulattri is LC50 = 0.349% and LC95 = 3.256% and that for P. aduncum is LC50 = 0.530% and LC95 = 4.666%. S. indicum oil (at 10% concentration) only caused the mortality of S. frugiperda by 27.5%. Insecticide mixture can increase the toxicity of the insecticide. The observation mortality of S. frugiperda for C. soulattri and P. aduncum (1:2) extracts mixture were LC50 = 0.233% and LC95 = 0.808%. At the same time, C. soulattri extract dan S. indicum oil mixture (4:1) were LC50 = 0.268% and LC95 = 0.931%. The treatments with a single insecticide and their mixtures affected the biological activity of S. frugiperda by reducing the area of feed consumption, and the longer the larval development time, the lower the pupal weight of S. frugiperda. Our findings indicated that a mixture of C. soulattri and P. aduncum extract, then C. soulattri extract, and S. indicum oil could potentially develop as effective insecticide for controlling S. frugiperda.

Keywords: biological disruption; botanical insecticides mixture; joint actions; synergism

1 Introduction

Spodoptera frugiperda is an invasive pest that attacks maize plants. In several areas in West Java, Indonesia, this pest attacks hybrid and sweet maize [1]. This pest was recorded as having 100 host plants [2]. S. frugiperda has superior characteristics such as surviving in various habitats, migrating well, having high fecundity, and fast resistance to insecticides [3]. The damage caused by S. frugiperda was high. The larvae are very damaging to the early stages of maize growth, and sixth instars larvae can cut the base of the maize seedlings until the plants die [4,5]. Attacks by S. frugiperda can cause yield losses of 8.3–20.6 million per ton [6].

An insecticide is a control technique that is easy to do, efficient, and effective. However, using high intensity and doses of insecticides can lead to the accumulation of insecticides and accelerate resistance in S. frugiperda [7]. Botanical insecticides are known to have various components that can delay pest resistance. The plant sources of botanical insecticides are very diverse. Afrianto et al. [8] showed 130 plant species with insecticidal abilities in Indonesia. Botanical insecticides have advantages over synthetic insecticides, such as broad spectrum in pest management, relatively low risk and nontoxic to natural enemies or nontarget organisms, and readily biodegradable, have varied modes of action, low persistence (leaving no residue on food or in the environment), and can be easily developed and applied. Therefore, botanical insecticides should be included in integrated pest management systems and contribute to sustainable agriculture [9–11].

Botanical insecticide source plants have been extensively researched and developed to control plant pests. Calophyllum soulattri and Piper aduncum are among the plants reported to have insecticidal properties against several pests. Meanwhile, Sesamum indicum was reported to have synergistic properties when mixed with insecticides. Volpe et al. [12] showed that essential oil of P. aduncum at a concentration of 85.4% dillapiole caused 100 and 98.75% mortality of nymphs and adult D. citri. Methanolic extract of C. soulattri stem bark caused the mortality of C. pavonana with an LC50 value of 0.15% [13]. This study tested the toxicity of C. soulattri, P. aduncum, and S. indicum and the potential of their mixtures to develop eco-friendly management strategies for controlling S. frugiperda.

2 Materials and methods

The experiment was carried out at the Pesticide and Environment Toxicology Laboratory, Department of Plant Pest and Diseases, Faculty of Agriculture, Universitas Padjadjaran, Jatinangor, West Java, Indonesia. The experiment was conducted at 27–32°C and humidity of 62–75%.

2.1 Test insect and botanical insecticide

2.1.1 Rearing of S. frugiperda

The insect test was S. frugiperda, already reared in the Laboratory of Pesticide and Environment Toxicology. The colony larvae were fed baby corn (pesticide-free), and the adults were fed 10% honey solution in cotton swabs.

2.1.2 Planting of maize plant

The leaves maize plant (Zea mays, a variety of Talenta) was used as the feed of S. frugiperda larvae in the test. Maize plant planting was done on a mixture of soil with manure (2:1). Maintenance of the plant included watering, weeding, and mechanical pest exclusion.

2.1.3 Insecticidal ingredients and extraction

C. soulattri stem bark was obtained from Teluk Melano District, Ketapang Regency, West Kalimantan Province, Indonesia. C. soulattri Stem bark extraction using ethanol solvent refers to the research by Syahputra [13]. The dried bark is mashed and soaked in ethanol (70% purity) at 1:10 ratio. Then, it was filtered using Whatman filter paper no.41. The solvent was evaporated by rotary evaporation. The resulting extract is stored in a bottle in the refrigerator (temperature 4℃) until use.

P. aduncum fruit was obtained from Masigit mountain, Padalarang Regency, West Java Province, Indonesia. Extraction of the fruit of P. aduncum was done using a methanol solvent. The fruit of P. aduncum was cut into small pieces and then dried at room temperature for 10 days (water content ±4.4%). After the P. aduncum fruit was dried, it was mashed with a blender, then immersed in methanol solvent (70% purity) with a ratio of 1:10 for 3 × 24 h, and macerated three times. Then it was filtered using Whatman filter paper no. 41. The filtered results were evaporated using a rotary evaporator at a temperature of 55–60°C and at a pressure of 300–500 mm Hg.

S. indicum was obtained from the market. S. indicum seeds are pressed to obtain oil using a seed press machine (MKS-J05). The oil is put in bottles and stored at room temperature.

2.2 Experiment bioassays

2.2.1 Single botanical insecticides test

The toxicity test of C. soulattri extract, P. aduncum extract, and S. indicum oil in the five-series concentration was based on a preliminary test and control. The solution concentration was dissolved in the methanol + acetone (1:4) (pro analysis). Control was prepared using methanol + acetone (1:4) (pro analysis). Bioassay was conducted by using a feeding method. Leaves cut with a length of 4 cm × 4 cm were given insecticides with a micro syringe (100 µl/leaf surface). Then, the leaf was air dried on the paper. Two pieces of treated leaves were put into a Petri dish line with filter paper. Then, 10 second instar of S. frugiperda larvae were put into a Petri dish using a fine brush.

The feeding treatment period was 48 h. Furthermore, larvae were fed with baby corn without treatment until they reached pupae, and each live larva was placed in a plastic container (50 ml). The observation was made by counting the number of dead larvae, larva development, the area of feed consumed, and the weight of pupae.

Observation:

The observation of mortality of S. frugiperda larvae was carried out from the day after treatment until 16 days after treatment. The mortality of larvae was calculated in the percentage of death by following equation (1).

(1) Mortality ( % ) = Σ D ead S . frugiperda Σ S . frugiperda tested × 100 % .

The observation of larvae development time was started from 1 day after treatment until instar VI larvae at 24 h observation interval. The observation was made by recording the time for the larvae to instar III, IV, V, and VI.

Feed consumption was observed by measuring the leaf area eaten by larvae using a transparent millimeter block. Next, the leaf area was calculated as the percentage by equation (2).

(2) Feed consumption ( % ) = Σ L eaf area eaten Σ Total leaf area × 100 % .

Observation of pupae weight was weighed using an analytic scale. The study arranges a randomized block design with four replications. Mortality test insects were analyzed probit with Polo Plus program 1.1. Feed consumption was analyzed by using analysis of variance. If the results were significant, then data were analyzed with the Duncan test using the SPSS 26 program. Then, data from larvae development time and pupae weight were analyzed descriptively.

2.2.2 Mixture of botanical insecticide test

The mixture of botanical insecticides test uses the method described in a single botanical insecticides test. The insecticide mixture used included a mixture of C. soulattri extract and P. aduncum extract at a ratio of 1:2, as well as a mixture of C. soulattri extract and S. indicum oil at a ratio of 4:1. The insecticide mixture activity analyzes based on different joint action models by calculating the combination index (CI) at the LC50 and LC95 levels [14] in equation (3).

(3) CI = LC x 1 ( cm ) LC x 1 + LC x 2 ( cm ) LC x 2 + LC x 1 ( cm ) LC x 1 × LC x 2 ( cm ) LC x 2 .

LC x 1 and LC x 2 are LC x insecticide 1 and insecticide 2 in separate tests, respectively, while LC x 1 ( cm ) dan LC x 2 ( cm ) are the respective lethal concentration (LC) components of the insecticide 1 and insecticide 2, respectively, in the mixture resulting in mortality (e.g., 50% and 95%). The LC value was obtained by multiplying the LC x of the mixture with the proportion of the concentration of component insecticide 1 and insecticide 2.

The categories of mixed activity properties are as follows:

CI < 0.5: strong synergies

CI 0.5–0.77: weak synergies

CI > 0.77–1.43: additive

CI > 1.43: antagonistic

Furthermore, the co-toxicity ratio of mixed insecticides with synergistic ingredients is calculated based on equation (4) [15]:

(4) Mixed co-toxicity ratio = LC x single insecticides LC x insecticides + S inergyst .

The categories of mixed activity properties are as follows:

CI < 1: antagonistic

CI > 1: synergies

CI = 1: no effect

3 Result

3.1 Effect of C. soulattri, P. aduncum, and S. indicum on mortality

Treatment with C. soulattri ethanol extract, P. aduncum methanol extract, and S. indicum oil to second instar S. frugiperda larvae showed various mortality responses according to the treatment. The mortality value of S. frugiperda larvae increased sharply on the second day after treatment (DAT). There was no significant increase in mortality the following day (Figures 1–3). The mortality trend of S. frugiperda shown in Figures 1 and 2 shows that the C. soulattri ethanol extract, P. aduncum methanol extract used, had a fast-acting method in killing the test insects. The results of the S. indicum oil test in Figure 3 shows that at a test concentration of 10%, the mortality of S. frugiperda only reached 27.5%. So no further tests were carried out to obtain the LC value. The test also confirms that S. indicum is a synergistic substance not toxic when applied alone.

Figure 1 Mortality of S. frugiperda after treatment with C. soulattri.
Figure 1

Mortality of S. frugiperda after treatment with C. soulattri.

Figure 2 Mortality of S. frugiperda after treatment with P. aduncum.
Figure 2

Mortality of S. frugiperda after treatment with P. aduncum.

Figure 3 Mortality of S. frugiperda after treatment with S. indicum.
Figure 3

Mortality of S. frugiperda after treatment with S. indicum.

The results of the probit analysis showed that the LC value decreased the more prolonged observation time. A decrease in the LC value of an insecticide indicates an increase in the toxicity value of the insecticide. The LC50 and LC95 values at the last observation (16 DAT) for the C. soulattri and P. aduncum tests were as follows: 0.349 and 3.256% (Table 1) and 0.530 and 4.666% (Table 2), respectively. If we look at the LC50 and LC95 values, the concentration values of the C. soulattri ethanol extract and P. aduncum methanol extract are high enough. However, the LC50 value is less than 0.6%. The LC value shows that insecticides are still less toxic to S. frugiperda.

Table 1

Probit analysis of the correlation of the concentration of C. soulattri insecticide and mortality of S. frugiperda

Day after treatment a ± SE b ± SE LC50 CI95 LC95 CI95
2 HSP 0.755 ± 0.143 1.880 ± 0.219 0.349 0.262–0.466 3.973 1.895–5.845
4 HSP 0.775 ± 0.141 1.696 ± 0.204 0.349 1.980–6.965 3.256 1.980–6.965
8 HSP 0.775 ± 0.141 1.696 ± 0.204 0.349 1.980–6.965 3.256 1.980–6.965
16 HSP 0.775 ± 0.141 1.696 ± 0.204 0.349 1.980–6.965 3.256 1.980–6.965

a – intercept; b – slope; SE – standard error; LC – lethal concentration (%); CI – confidence interval.

Table 2

Probit analysis of the correlation of the concentration of P. aduncum insecticide and mortality of S. frugiperda

Waktu pengamatan a ± SE b ± SE LC50 CI95 LC95 CI95
2 HSP 0.310 ± 0.114 1.838 ± 0.219 0.679 0.521–0.889 5.325 3.346–10.805
4 HSP 0.386 ± 0.116 1.826 ± 0.217 0.615 0.471–0.805 4.895 3.087–9.864
8 HSP 0.480 ± 0.117 1.741 ± 0.212 0.530 0.400–0.698 4.666 2.894–9.730
16 HSP 0.480 ± 0.117 1.741 ± 0.212 0.530 0.400–0.698 4.666 2.894–9.730

a – intercept; b – slope; SE – standard error; LC – lethal concentration (%); CI – confidence interval.

The results of mixed insecticide tests showed that mixing C. soulattri ethanol extract with P. aduncum methanol extract and C. soulattri ethanol extract with S. indicum oil increased the toxicity of the insecticides. The increase in toxicity occurs if the comparison between the ingredients is in the right amount. The test results showed that at a concentration of 1%, the mixed insecticides could cause up to 100% mortality (Figure 4). After further testing, the mortality of the test insects is shown in Figures 5 and 6. The mortality trend showed that insecticide toxicity occurred at the beginning of the observation (2–4 days after treatment), and there was no increase in the mortality of the test insects on the next day. The LC50 and LC95 values of the mixed insecticides (Tables 3 and 4) were lower than the single insecticides (Tables 1 and 2). This experiment shows that mixing can increase the toxicity of insecticides more toxic than a single application.

Figure 4 Mortality of S. frugiperda larvae at 5 days after treatment with a mixture of botanical insecticides. PACS – P. aduncum and C. soulattri; CSSI – C. soulattri and S. indicum; PASI – P. aduncum and S. indicum; PA – P. aduncum; SI – S. indicum; CA – C. soulattri.
Figure 4

Mortality of S. frugiperda larvae at 5 days after treatment with a mixture of botanical insecticides. PACS – P. aduncum and C. soulattri; CSSI – C. soulattri and S. indicum; PASI – P. aduncum and S. indicum; PA – P. aduncum; SI – S. indicum; CA – C. soulattri.

Figure 5 Mortality of S. frugiperda after treatment with mixture C. soulattri and P. aduncum (1:2). CSPA – C. soulattri + P. aduncum (1:2).
Figure 5

Mortality of S. frugiperda after treatment with mixture C. soulattri and P. aduncum (1:2). CSPA – C. soulattri + P. aduncum (1:2).

Figure 6 Mortality of S. frugiperda after treatment with mixture C. soulattri and S. aduncum (4:1). CSSI – C. soulattri + S. indicum (4:1).
Figure 6

Mortality of S. frugiperda after treatment with mixture C. soulattri and S. aduncum (4:1). CSSI – C. soulattri + S. indicum (4:1).

Table 3

Probit analysis of the correlation of the concentration of C. soulattri and S. indicum mixture insecticide and mortality of S. frugiperda

Day after treatment a ± SE b ± SE LC50 CI50 LC95 CI95
2 HSP 2.004 ± 0.252 3.289 ± 0.386 0.246 0.210–0.286 0.778 0.607–1.127
4 HSP 1.976 ± 0.249 3.169 ± 0.375 0.238 0.202–0.278 0.786 0.570–1.368
8 HSP 0.1.927 ± 0.245 3.049 ± 0.365 0.233 0.197–0.274 0.808 0.620–1.209
16 HSP 0.1.927 ± 0.245 3.049 ± 0.365 0.233 0.197–0.274 0.808 0.620–1.209

a – intercept; b – slope; SE – standard error; LC – lethal concentration (%); CI – confidence interval.

Table 4

Probit analysis the of correlation of the concentration of C. soulattri and P. aduncum mixture insecticide and mortality of S. frugiperda

Day after treatment a ± SE b ± SE LC50 CI50 LC95 CI95
2 HSP 1.740 ± 0.223 3.042 ± 0.366 0.268 0.226–0.315 0.931 0.715–1.388
4 HSP 1.740 ± 0.223 3.042 ± 0.366 0.268 0.226–0.315 0.931 0.715–1.388
8 HSP 1.740 ± 0.223 3.042 ± 0.366 0.268 0.226–0.315 0.931 0.715–1.388
16 HSP 1.740 ± 0.223 3.042 ± 0.366 0.268 0.226–0.315 0.931 0.715–1.388

a – intercept; b – slope; SE – standard error; LC – lethal concentration (%); CI – confidence interval.

The joint action of the first mixture (C. soulattri and P. aduncum extract) was determined based on the independent joint action model, where both ingredients have an insecticidal effect. The second mixture has a synergistic joint action (C. soulattri extract and S. indicum oil). The ingredients can increase toxicity in the mixed state, and if in a single application, they are not toxic. The results of the interaction of mixed insecticides can be in the form of synergism (the mixed effect is more excellent than expected) and antagonism (the mixtures interfere with each other’s effects) [16]. Analysis of the CI or activity of the mixture also showed that the C. soulattri extract with S. indicum oil (4:1) and C. soulattri with P. aduncum extract (1:2) mixture were synergistic at LC95 value. Joint action at LC50 value shows that the mixture of C. soulattri and P. aduncum extract was antagonistic (Table 5). However, compared to the LC value, the LC50 value of mixed insecticides is lower or more toxic. Comparison of the ratio between insecticides can modify to increase the toxicity of insecticides.

Table 5

Combination index of insecticides mixture

Treatment Combination index Combination index
2 HSP 16 HSP 2 HSP 16 HSP
LC50 LC95 LC50 LC95 LC50 LC95 LC50 LC95
CSPA 1.466 0.450 1.662 0.543 Antagonistic Strong synergistic Antagonistic Weak synergistic
CSSI 1.419 5.107 1.498 4.030 Synergistic Synergistic Synergistic Synergistic

CSPA – C. soulattri + P. aduncum (1:2); CSSI – C. soulattri + S. indicum (4:1); LC – lethal concentration (%).

3.2 Effect of C. soulattri, P. aduncum, and S. indicum on development time

Treatment using C. soulattri ethanol extract, P. aduncum methanol extract, and S. indicum oil can inhibit the long development of S. frugiperda if compared to controls. Treatment of C. soulattri with a concentration of 2.5% could inhibit larval development by extending the larval development time by 0.7 days (instars II–VI) (Table 6). The treatment of P. aduncum and S. indicum extended the development time (II–VI instars) by 1.12 and 0.04 days, respectively (Tables 7 and 8). The mixed insecticide treatment also showed that the insecticide could prolong the development time of the S. frugiperda larvae. The highest concentration of C. soulattri extract and S. indicum oil (4:1) mixture could prolong the larval development time from instars II-VI by 1.05 days, while the C. soulattri and P. aduncum (1:2) extract mixture could prolong development time by 2.77 days (Tables 9 and 10).

Table 6

Length of development time of S. frugiperda larvae after treatment with C. soulattri

Treatment Length of larval development at test concentration (X ± SE) (days)
N Instar II–II N Instar II–IV N Instar II–V N Instar II–VI
CS (2.5%) 2 3.50 ± 0.354 2 5.50 ± 0.354 2 9.50 ± 0.354 2 12.50 ± 0.354
CS (0.94%) 11 3.18 ± 0.116 11 5.91 ± 0.239 11 10.00 ± 0.223 11 12.36 ± 0.145
CS (0.35%) 22 3.05 ± 0.044 20 5.60 ± 0.130 20 9.35 ± 0.146 20 12.45 ± 0.111
CS (0.13%) 34 3.44 ± 0.085 30 5.63 ± 0.120 30 9.33 ± 0.109 30 12.37 ± 0.100
CS (0.05%) 39 3.18 ± 0.102 37 5.54 ± 0.082 37 9.14 ± 0.116 37 12.41 ± 0.081
Control 40 2.45 ± 0.100 40 5.48 ± 0.079 40 8.75 ± 0.116 40 11.88 ± 0.052

CS – C. soulatrd, X – average of the length development time (day), SE – Standard Error, N – number of larvae.

Table 7

Length of development time of S. frugiperda larvae after treatment with P. aduncum

Treatment Length of larval development at test concentration (X ± SE) (days)
N Instar II–II N Instar II–IV N Instar II–V N Instar II–VI
PA (3.9%) 4 4.75 ± 0.216 2 6.50 ± 0.354 2 9.50 ± 0.000 2 11.50 ± 0.000
PA (1.56%) 11 4.09 ± 0.087 7 6.43 ± 0.187 7 9.43 ± 0.114 7 11.43 ± 0.126
PA (0.625%) 22 3.50 ± 0.107 21 6.10 ± 0.177 21 8.90 ± 0.091 21 11.43 ± 0.105
PA (0.25%) 34 3.44 ± 0.085 30 5.90 ± 0.136 30 8.57 ± 0.075 30 11.60 ± 0.083
PA (0.1%) 36 3.33 ± 0.079 36 5.86 ± 0.119 36 8.47 ± 0.078 36 11.53 ± 0.075
Control 40 2.63 ± 0.121 40 5.30 ± 0.072 40 8.38 ± 0.116 40 10.38 ± 0.052

PA – P. aduncum; X – average of the length development time (day), SE – standard error, N – number of larvae.

Table 8

Length of development time of S. frugiperda larvae after treatment with S. indicum

Treatment Length of larval development at test concentration (X ± SE) (days)
N Instar II–II N Instar II–IV N Instar II–V N Instar II–VI
SI (10%) 30 3.67 ± 0.088 29 6.30 ± 0.143 29 9.66 ± 0.112 29 11.34 ± 0.088
SI (7.07%) 31 3.26 ± 0.079 30 6.32 ± 0.140 30 9.60 ± 0.101 30 11.43 ± 0.090
SI (4.10%) 33 3.42 ± 0.086 33 6.15 ± 0.136 33 9.30 ± 0.091 33 11.31 ± 0.082
SI (3.54%) 34 3.32 ± 0.080 34 6.18 ± 0.147 34 9.24 ± 0.073 34 11.18 ± 0.065
SI (2.5%) 35 3.31 ± 0.078 35 6.09 ± 0.130 35 9.29 ± 0.076 35 11.50 ± 0.083
Control 40 2.33 ± 0.089 40 5.45 ± 0.079 40 8.73 ± 0.117 40 11.30 ± 0.072

SI – S. indicum, X – average of the length development time (days), SE – Standard Error, N – number of larvae.

Table 9

Length of development time of S. frugiperda larvae after treatment with C. soulattri and S. indicum mixture (4:1)

Treatment Length of larval development at test concentration (X ± SE) (days)
N Instar II–III N Instar II–IV N Instar II–V N Instar II–VI
CSSI 0.09% 36 3.61 ± 0.098 36 5.81 ± 0.151 36 8.83 ± 0.062 36 10.83 ± 0.062
CSSI 0.16% 27 3.70 ± 0.088 27 5.96 ± 0.161 27 8.70 ± 0.088 27 10.70 ± 0.088
CSSI 0.285% 11 3.18 ± 0.116 11 5.64 ± 0.194 11 9.00 ± 0.182 11 10.91 ± 0.155
CSSI 0.506% 7 3.57 ± 0.187 7 6.00 ± 0.286 7 9.00 ± 0.202 7 10.71 ± 0.265
CSSI 0.90% 1 4.00 ± 0.078 1 6.00 ± 0.000 1 10.00 ± 0.00 1 11.00 ± 0.000
Control 40 2.25 ± 0.068 40 4.78 ± 0.066 40 8.08 ± 0.108 40 10.05 ± 0.034

CSSI – C. soulattri + S. indicum (4:1), X – average of the length development time (day), SE – standard error, N – number of larvae.

Table 10

Length of development time of S. frugiperda larvae after treatment with C. soulattri and P. aduncum mixture (1:2)

Treatment Length of larval development at test concentration (X ± SE) (days)
N Instar II–III N Instar II–IV N Instar II–V N Instar II–VI
CSPA 0.11% 35 2.46 ± 0.084 34 5.32 ± 0.090 34 8.41 ± 0.119 34 11.00 ± 0.000
CSPA 0.195% 27 3.00 ± 0.000 27 6.15 ± 0.068 27 9.30 ± 0.088 27 11.19 ± 0.075
CSPA 0.348% 14 3.00 ± 0.000 14 6.93 ± 0.213 14 9.57 ± 0.132 14 11.57 ± 0.132
CSPA 0.618% 6 3.00 ± 0.000 6 7.50 ± 0.312 6 9.67 ± 0.192 6 11.50 ± 0.204
CSPA 1.10% 1 3.00 ± 0.000 1 7.00 ± 0.000 1 10.00 ± 0.000 1 12.00 ± 0.000
Control 40 2.20 ± 0.089 40 5.43 ± 0.078 40 8.43 ± 0.078 40 9.23 ± 0.090

CSPA – C. soulattri + P. aduncum (1:2), X – average of the length development time (day), SE – standard error, N – number of larvae.

3.3 Effect of C. soulattri, P. aduncum, and S. indicum on feed consumed

In addition, C. soulattri ethanol extract, P. aduncum methanol extract, and S. indicum oil and their mixture suppressed larvae consumption, which was lower than the control. The statistical analysis also showed that the treatments differed significantly from the control (Tables 11 and 12). The test results showed that the single and mixed insecticides tested could reduce feed consumption to reduce crop damage due to attacks by S. frugiperda larvae.

Table 11

Leaf area consumed by S. frugiperda larvae after treatment with C. soulattri, P. aduncum, and S. indicum

Treatment The leaf area consumed (X ± SE) (%) Treatment The leaf area consumed (X ± SE) (%) Treatment The leaf area consumed (X ± SE) (%)
CS (2.5%) 0.703 ± 0.1213a PA (3.9%) 0.95 ± 0.14a SI (10%) 6.51 ± 0.0511a
CS 0.94%) 0.898 ± 0.1202a PA (1.56%) 1.98 ± 0.11b SI (7.07%) 7.65 ± 0.2523b
CS (0.35%) 2.555 ± 0.2177b PA (0.625%) 9.27 ± 0.26c SI (4.10%) 9.52 ± 0.2886c
CS (0.13%) 2.922 ± 0.1408b PA (0.25%) 12.23 ± 0.15d SI (3.54%) 13.18 ± 0.1840d
CS (0.05%) 5.156 ± 0.1154c PA (0.1%) 13.48 ± 0.26e SI (2.5%) 16.30 ± 0.2432e
Control 9.813 ± 0.3164d Control 17.62 ± 0.16f Control 17.35 ± 0.3004f

The average number followed by the same letter in the same column is not significantly different according to the Duncan test at 5% level; X – average of the leaf area consumed (%); SE – standard error; CS – C. soulattri; PA – P. aduncum; SI – S. indicum.

Table 12

Leaf area consumed by S. frugiperda larvae after treatment with C. soulattri, P. aduncum, and S. indicum mixture

Treatment The leaf area consumed (X ± SE) (%) Treatment The leaf area consumed (X ± SE) (%)
CSSI 0.09% 6.328 ± 0.505161c CSPA 0.11% 14.188 ± 1.123317b
CSSI 0.16% 2.602 ± 0.489062b CSPA 0.195% 10.008 ± 0.931254ab
CSSI 0.285% 1.672 ± 0.508413ab CSPA 0.348% 6.305 ± 0.962072ab
CSSI 0.506% 0.984 ± 0.218331ab CSPA 0.618% 1.508 ± 0.193704a
CSSI 0.90% 0.547 ± 0.132813a CSPA 1.10% 0.414 ± 0.043322a
Control 12.813 ± 0.508998d Control 38.891 ± 1.066742c

The average number followed by the same letter in the same column is not significantly different according to the Duncan test at 5% level; CSSI – C. soulattri + S. indicum (4:1); CSPA – C. soulattri + P. aduncum (1:2); X – average of the leaf area consumed (%); SE – standard error.

3.4 Effect of C. soulattri, P. aduncum, and S. indicum on pupae weight

Insecticide application made the pupal weight of S. frugiperda lower than the control. The treatment disrupts the physiological processes of pests until it affects the weight of the pupae. However, the S. indicum treatment did not reduce the pupal weight more than the control. Observations on the pupal weight are presented in Tables 13 and 14.

Table 13

Average weight of S. frugiperda pupae after treatment with C. soulattri, P. aduncum, and S. indicum

Treatment The weight of the pupae (X ± SE) (%) N Treatment The weight of the pupae (X ± SE) (%) N Treatment The weight of the pupae (X ± SE) (%) N
CS (2.5%) 0.2108 ± 0.01471 2 PA (3.9%) 0.1911 ± 0.01064 2 SI 10% 0.2280 ± 0.00347 29
CS 0.94%) 0.2077 ± 0.00538 11 PA (1.56%) 0.1990 ± 0.00553 7 SI 7.07% 0.2120 ± 0.00644 30
CS (0.35%) 0.2022 ± 0.00478 20 P A (0.625%) 0.1942 ± 0.00657 21 SI (4.10% 0.2075 ± 0.00315 33
CS (0.13%) 0.2029 ± 0.00266 30 PA (0.25%) 0.1890 ± 0.00199 30 SI (3.54% 0.2234 ± 0.00213 34
CS (0.05%) 0.2017 ± 0.00413 37 PA (0.1%) 0.2003 ± 0.00417 36 SI 2.5% 0.2115 ± 0.00242 35
Control 0.2111 ± 0.00185 40 Control 0.2128 ± 0.00571 40 Control 0.2180 ± 0.00298 40

CS – C. soulattri; PA – P. aduncum; SI – S. indicum, X – average of the weight the pupae (g); SE – standard error, N – number of pupae.

Table 14

Average weight of S. frugiperda pupae after treatment with C. soulattri, P. aduncum, and S. indicum mixture

Treatment The weight of the pupae (X ± SE) (%) N Treatment The weight of the pupae (X ± SE) (%) N
CSSI 0.09% 0.2462 ± 0.00000 36 CSPA 0.11% 0.2100 ± 0.00467 34
CSSI 0.16% 0.2253 ± 0.00297 27 CSPA 0.195% 0.2171 ± 0.00731 27
CSSI 0.285% 0.2224 ± 0.00208 11 CSPA 0.348% 0.2165 ± 0.00412 14
CSSI 0.506% 0.2192 ± 0.00269 7 CSPA 0.618% 0.2125 ± 0.00628 6
CSSI 0.90% 0.2176 ± 0.00000 1 CSPA 1.10% 0.1992 ± 0.00000 1
Control 0.2351 ± 0.00265 40 Control 0.2184 ± 0.00180 40

CSSI – C. soulattri + S. indicum (4:1); CSPA – C. soulattri + P. aduncum (1:2); X – average of the weight the pupae (g); SE – standard error; N – number of pupae.

4 Discussion

The result obtained from the present study demonstrated the potential effect of insecticide from C. soulattri ethanol extract, P. aduncum methanol extract, and a mixture of C. soulattri extract with P. aduncum extract and a mixture of C. soulattri extract and S. indicum oil for controlling S. frugiperda. Apart from causing death, applying insecticides also disrupted the biological activity of S. frugiperda. These disturbances are suppression of feeding activity, lower pupal weights, and prolonged development time. In addition, this study demonstrated the potential for using this insecticide mixture as an alternative strategy to manage this important pest species.

The main effects observed were on the mortality of S. frugiperda. The mortality effect was due to the presence of insecticide ingredients from the extract. Dillapiole from P. aduncum is an active compound that causes the mortality of test insects [17,18]. Other main compound components of P. aduncum include myristicin, aromadendrene, dillapiole, α-serine, tridecane, γ-elemene, o-cymene, Z-carpacin [19,20], piperitone, terpinen-4-ol, β-caryophyllene, α-humulene, and germacrene-D [21]. C. soulattri extract contains saponins, flavonoids, terpenoids, steroids, phenols, and tannins [21]. C. soulattri mode of action on the active fraction was reported to be faster and did not show symptoms of hormone disruption in insect development [13].

Among all botanical insecticides tested and based on LC value, a mixture of C. soulattri extract with P. aduncum extract (1:2) and a mixture of C. soulattri extract and S. indicum oil (4:1) were more toxic compared to a single application (Tables 1–4). Mixing insecticides is more effective because insect pests encounter a complex mixture of secondary compounds during the feeding process [22]. Scott et al. [23] stated that Piper spp. has a methylenedioxyphenyl group that can synergize when mixed with other plant extracts. S. indicum inhibits the action of P450s and potentiates insecticidal activity [24]. The combination of active compounds increases the insecticide spectrum, lowers residues, and reduces harmful effects on the environment and nontarget organisms [25].

The results of the calculation of the CI of the two mixtures used in this study showed synergistic properties at the LC95 value. However, at the LC50 value, the mixture of P. aduncum and C. soulattri extracts was antagonistic, while the mixture of C. soulattri extract and S. indicum oil was synergistic (Table 5). Several comparisons between ingredients can be made to optimize the toxicity and synergistic effect of the mixture used. Some reports show that insecticide mixtures with different ratios showed different effects. The study by Susanto and Prijono [26] on P. aduncum and Tephrosia vogelii at a ratio of 1:2 showed strong synergy at LC50 and weak synergy at LC95 against Scirpophaga incertulas. Another mixture between P. aduncum and Sapindus rarak (1:10) is synergistic at LC95 but additive at LC50 [27]. Nailufar and Prijono [28] demonstrated that a mixture of P. aduncum and T. vogellli in the ratios of 1:5, 1:1, and 5:1 was 2.4, 2.5, and 3.4 times more toxic than T. vogelli single extract, respectively. Furthermore, a mixture of S. indicum with clove oil (8:2) treatment can increase the mortality of Callosobruchus maculatus compared to single clove treatment alone [29]. This explanation illustrates that the ratio between the ingredients in the mixture determines the combination index for insecticidal and mixed activity properties.

Other effects observed in this study were development time (Tables 6–10), area of feed consumption (Tables 11 and 12), and pupal weight (Tables 13 and 14). The results showed the application of insecticides C. soulattri ethanol extract, P. aduncum methanol extract, a mixture of C. soulattri with P. aduncum extract, and a mixture of C. soulattri extract and S. indicum oil affected these parameters. The development of larvae exposed to insecticides used slowed the development time. This happened because of disturbances in the physiology and biochemistry of S. frugiperda due to insecticide exposure. The energy that should be used for growth and development is used to detoxify insecticides in the bodies. Then the observation parameter of consumption area emphasizes the area of feed consumption, which shows the antifeedant properties of the test insecticides used. C. soulattri is reported to have a function as a primary antifeedant (rejection to eat) and secondary antifeedant (affects the process of eating, digestion, and absorption) [30]. Besides that, S. indicum ligands (sesamol and pinoresinol) provide antifeedant effects and juvenile hormone analogues [31,32]. Research by Cossolin et al. [33] stated that treatment of P. aduncum to Euschistus heros caused cytological changes such as tissue disruption, increased mitochondrial population, and depletion of glycogen and lipids in the body’s fat cells. The presence of antifeedant properties in C. soulattri and P. aduncum extracts and S. indicum oil allows for a combination of actions that can enhance the antifeedant effect. The low weight is likely due to the antifeedant properties of the tested insecticide’s effect on feeding activity and consumption rates decrease [34]. Other studies have also shown that the components of botanical insecticides affect feed consumption and the deficiency of nutrients for larvae to support their growth [35]. However, at sub-LCs, botanical insecticides did not affect reducing the weight of the test insects [36].

These results indicate that S. frugiperda larvae when exposed to insecticide tests, not only kill the test insects but cause biological disruption in S. frugiperda. This effect can contribute to an integrated management strategy to keep the pest population below the damage threshold. Therefore, insecticides derived from C. soulattri, P. aduncum, and their mixture with S. indicum can serve as more environmentally friendly control alternatives for managing S. frugiperda.

5 Conclusion

Applying single and mixed S. soulattri ethanol extract, P. aduncum methanol extract, and S. indicum oil caused mortality in S. frugiperda. LC50 and LC95 value in the insecticide mixture is more toxic than LC50 and LC95 in a single application. The mixture used in this test was C. soulattri ethanol extract and P. aduncum methanol extract (1:2), and then a mixture of C. soulattri ethanol extract and S. indicum oil (4:1) was used. The CI of insecticide mixture shows that at the LC95 value, the mixture has a synergistic effect. The botanical insecticides in this test showed a biological disturbance in S. frugiperda by reducing the area of feed consumption, prolonging the larval development time, and reducing the weight of S. frugiperda pupae. Both mixtures can be developed in eco-friendly management strategies for the economic and sustainable management of S. frugiperda in maize plants.

  1. Funding information: This research was funded by the Internal Grant Universitas Padjadjaran with the Unpad lecturer competency research scheme in 2022 (No. 2203/UN6.3.1/PT.00/2022) and continues funded by Academic Leadership Grant scheme in 2023 (No. 1549/UN6.3.1/PT.00/2023).

  2. Conflict of interest: The authors state no conflict of interest.

  3. Data availability statement: The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.

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Received: 2023-04-05
Revised: 2023-05-09
Accepted: 2023-06-30
Published Online: 2023-09-20

© 2023 the author(s), published by De Gruyter

This work is licensed under the Creative Commons Attribution 4.0 International License.

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  86. Breakfast habits and knowledge: Study involving participants from Brazil and Portugal
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  89. Special Issue on Agriculture, Climate Change, Information Technology, Food and Animal (ACIFAS 2020)
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  91. Enhancing the ability of rice to adapt and grow under saline stress using selected halotolerant rhizobacterial nitrogen fixer
https://doi.org/10.1515/opag-2022-0213
Keywords for this article
biological disruption; botanical insecticides mixture; joint actions; synergism
Creative Commons
BY 4.0

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