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Journal of Bioremediation & Biodegradation
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  • Research   
  • JBRBD, Vol 14(1)

Investigation of Plants Extracts as Potential Larvicidal Agents against Culex quinquefasciatus and Anopheles stephensi

Rajalakshmi1 and Jayalakshmi Krishnan2*
1Department of Life Sciences, Central University of Tamil Nadu, Thiruvarur, India
2Departments of Life Sciences, Central University of Tamil Nadu, Thiruvarur, India
*Corresponding Author: Jayalakshmi Krishnan, Department of Life Sciences, Central University of Tamil Nadu, Thiruvarur, India, Email: jayalakshmi@cutn.ac.in

Manuscript No. jbrbd-23-85534 / Editor assigned: 05-Jan-2023 / PreQC No. jbrbd-23-85534 (PQ) / Reviewed: 19-Jan-2023 / QC No. Jbrbd-23-85534 / Revised: 23-Jan-2023 / Manuscript No. Jbrbd-23-85534 (R) / Accepted Date: 30-Jan-2023 / Published Date: 30-Jan-2023

Abstract

To eradicate insects chemical insecticides have been used for long time. Synthetic insecticides have created havoc in the environment by altering the environment as well proved toxic no non-target animals/organisms. Hence, the focus to control the insects is now shifted to naturally occurring plant based insecticides which are eco-friendly. Among the insects, mosquitoes form important vectors as they transmit diseases like malaria and lymphatic filariasis etc. The present study is carried out to investigate the larvicidal activity of four different plants against Culex quinquefasciatus (vector of lymphatic filariasis) and Anopheles stephensi (malaria). There are many studies which state that the plant crude extracts act against mosquito as larvicidal, pupicidal, and adulticidal agents. As plants have antioxidant, toxic and bioactive properties they are used extensively in mosquito control. The chosen plants in this study are Ipomoea carnea (Leaves), Commiphora caudate (Leaves), Euphorbia ant quorum (Latex) and Acapulco senna-alata (Leaves). Ethanol, Methanol and Acetone extracts from four plants were tested against late third instar larvae of Culex quinquefasciatus and Anopheles stephensi. The larvae were exposed to different concentrations of plant crude extracts. After 24 hours the mortality rate of each dose was observed. There was no mortality in control. Among the plants screened, Ipomoea carnea (Leaves), crude extract showed highest mortality (100%), followed by Commiphora caudate (Leaves) which is 97%. The other two plants such as Euphorbia antiquorum and Acalpulco senna-alata showed 86% and 78% mortality respectively. The chemical composition of plant crude extract were analyzed by GC-MS and given herewith. The plant extracts needs to be tested for further as an effective larvicidal under field conditions.

Keywords

Plants; Mosquito Larvicidal activity; GC-MS; Lethal concentration

Introduction

Vector borne diseases account for 17% of all the diseases among the various illnesses that kills the people. Blood sucking insects usually act as vectors to transmit the pathogens from human to human or from animals to humans. Mosquitoes form the major part of vector species others being the ticks, mites, sandflies, flea and bugs [1].Mosquitoes transmit a number of diseases, such as filariasis, dengue, malaria, yellow fever, chikungunya, Japanese encephalitis [2, 3]. Mosquitoes are also described as most dangerous animal in the world because of its ability to transmit malaria. There are 3,500 species of mosquitoes in earth [3]. Mosquitoes that transmit malaria are grouped as Anopheles. There are about 40 different species of Anopheles mosquitoes which transmit malaria [4]. Malaria parasites Plasmodium falciparum and Plasmodium vivax are carried by Anopheles stephensi important mosquito vector. This species of mosquito is abundant in Middle East, Indian subcontinent and also china [5]. Malaria is a serious global health problem that affect millions of people particularly young children (below 5 years old), pregnant ladies, and patients with conditions –like HIV/AIDS [6]. Lymphatic filariasis is a global health problem found mainly in tropical region of the world [6]. About 1.4 billion people in 73 countries of the world are threatened by the disease [7]. Lymphatic filariasis is called as Elephantiasisis a painful and disfiguring disease. Wuchereria bancrofti, Brugia malayi and Brugia timori, are the three types of nematode worms which cause the disease filariasis [8]. The vectors of genus Culex carry Wuchereria Bancroft, in urban and semi-urban areas, Aedes in pacific islands, Anopheles in Africa and elsewhere. Culex quinquefasciatus (Diptera Culicidae) is a predominant house-resting mosquito in most tropical countries [9]. This mosquito’s breed in polluted waters such as septic tanks, blocked drains, soakage pools close to human habitations [10]. There are scientific reports which say that the larvicidal activity can be seen in the plant extract from leaves and latex [11]. Repellent plant crude extract in mosquito control programs due to their excellent larvicidal, pupicidal and adulticidal properties have been carried out by various groups of scientists in recent past [12]. Plants have bioactive phytochemical compounds which act as source of alternative agents that control mosquito [13]. In our study, we have chosen Commiphora caudate, Ipomoea carnea, Euphorbia antiquorum, Acalpulco sennaalata as potential bioactive plants which act against Anopheles stephensi and Culex quinquefasciatus. However, to be noted is 80% of the words population use plant as their primary and secondary source of medication [14]. Ipomoea carnea (Convolvulaceae family) is the large diffuse shrub and it is grown in wetland areas and river beds [15]. Ipomoea carnea leaves contain seven compound such as stearic acid, 3-diethylamino-1-propanol, hexatriacontane, 1,2diethylphthalate, n-octadecanol, octacosane, hexadecanoic acid, tetracontane. This plant is used in Ayurveda traditional medicinal systems [16]. Euphorbia antiquorum (Euphorbiaceous family) is a medium size annual herb common to the tropical and sub-tropical areas. It grows 12 cm tall. Euphorbia antiquorum leaves and stem produce milky juice or white latex when cut. It’s grown in grassland, road sides and pathways etc. [17].Euphorbia antiquorum exhibit the antifungal, antimutagenic, antibacterial, insecticidal, molluscicidal, antitumor activity [18]. It generally grows in dry forest. Acalpulco senna-alata is a shrub. The leaves include chrysophanic acid [19]. It is commonly used for Skin disease, Tinea, Ringworm and athlete’s foot diseases. This plant is grown in many areas in south India [20]. Based on the above said information; the objective of the present study was to examine larvicidal activity of different plant extracts against Culex quinquefasciatus and Anopheles stephensi.

1. To identify and select the toxic plants and to find out the active compounds in the plants chosen

2. To rear the Anopheles stephensi and Culex quinquefasciatus mosquitoes in lab

3. To evaluate the larvicidal properties of different plant extracts against Anopheles stephensi and Culex quinquefasciatus.

Material and Methods

Plants Collection

The leaves of Commiphora caudate, Ipomoea carnea, Acalpulco senna-alata and latex of Euphorbia antiquorum were collected from the rural area of Maruthappatinum in Thiruvarur District, Tamil Nadu, given in [Figure 1a, 1b] and [Figure 1c, 1d].

image

Preparation of Ipomoea carnea extract

The Ipomoea carnea plant leaves were collected and the leaves were dried 7-10 days. The dried leaves (100g) were powdered using electrical blender. Powder was dissolved in 250ml of ethanol into 500ml of conical flask for 24 hrs. The mixture was filtered through whatmanNo.1 filter paper using Buchner funnel and then concentrated vacuum using rotary evaporator (400C) [Figure 2a]. The obtained extracts were kept at 40C until further use [13, 11].

bioremediation-biodegradation-carnea

Figure 2 (a): Ipomoea carnea leaf extraction.

Preparation of Commiphora caudate extract

The Commiphora caudate plant leaves were collected and then dried 7-10 days under room temperature. Then the dried leaves (100g) were powdered using electrical blender. The powdered plant materials were soaked in the extracting 500ml methanol solvents for 24 hrs. The mixture was filtered through whatmanNo.1 filter paper using Buchner funnel and then concentrated vacuum using rotary evaporator (400C) [Figure 2b]. The obtained extracts were kept at 40C until further use [4, 9].

bioremediation-biodegradation-Commiphora

Figure 2 (b): Commiphora caudate leaf extract.

Preparation of Euphorbia antiquorum extract

The Euphorbia antiquorum latex were collected. The collected latex was mixed with methanol in the ratio of 1:9 (10%) and centrifuged at 3500rpm for 5 minutes. The supernatant was collected in a glass vial and stored at 40C till further use [12, 18] shown in [Figure 2c].

bioremediation-biodegradation-antiquorum

Figure 2 (c): Euphorbia antiquorum latex extract.

Preparation of Acalpulcosenna-alata extract

The Acalpulcosenna-alata plant leaves were collected. The leaves of plant (100g) were air dried and ground to powder using grinder. The powdered plant material was extracted by maceration with shaking for 24 hrs, in 70% acetone with 10:1 solvent to dry weight ratio. The extract was the filtered through WhatmanNo.1 filter paper using Buchner funnel, and the acetone removed under steam of air. The acetone extract was dried in a rotary evaporator under reduced pressure and kept temperature 400C [30, 21] shown in [Figure 2d]. Buchner Funnel & Rotary Evaporator was used for all the plant extracts [Figure 3a, 3b].

bioremediation-biodegradation-Acalpulcosenna

Figure 2(d): Acalpulcosenna-alata leaf extraction

bioremediation-biodegradation-Buchner

Figure 3 (a): Buchner Funnel

bioremediation-biodegradation-Rotary

Figure 3(b): Rotary Evaporator

Rearing of larvae

The Mosquito eggs were collected from Centre Research in Medical Entomology (CRME), Madurai, Tamil Nadu State and Eggs were reared under laboratory condition from the Central University of Tamil Nadu, Thiruvarur. The larvae of Anopheles stephensi and Culex quinquefasciatus were feed with dog biscuit mixed with yeast powdered in 3:1 ratio were floated on the water surface after the larvae turned into the late third instar they were used for the bioassay [22].

Larvicidal Bioassay:

The larvicidal plant crude extract was evaluated as per the protocol previously described by WHO (2005). From the plant crude extract stock solutions six different test concentrations. (50,100,150,200,250 and 300 ppm) were prepared and tested against the freshly (24 hrs.) late third instar larvae of Anopheles stephensi and Culex quinquefasciatus as shown in [Figure 4]. The test medium (350ml paper cups) were used by adding 500μl of plant extract mixed with 99.5ml of distilled water to make up (50 ppm) of test sample.1000μl of plant crude extract mixed with 99.0ml of distilled water to make up (100 ppm) of test sample.1500μl of plant extract mixed with 98.5ml of distilled water to make up (150ppm) of test sample. 2000μl of plant crude extract mixed with 98.0ml of distilled water to make up (200ppm) of test sample.2500μl of plant crude extract mixed with 97.5ml of distilled water to make up (250ppm) of test sample.3000μl of plant crude extract mixed with 97.0ml of distilled water to make up (300ppm) of test sample. The control (with the plant extracts) experiments were also run parallel with each replicate. Four experiment; four replicates were maintained at a time. A minimum number of 25 larvae mortality were observed and recorded after 24hrs post treatment, percentage of mortality were calculated using Abbott’s formula. There is no mortality found in the control [23].

bioremediation-biodegradation-Larvicidal

Figure 4: Larvicidal Bioassay.

Abbots formula= % test mortality - %control mortality ×100

100 - %control mortality

Gas chromatography Mass spectrometry for plant compounds identification

All plant extracts (500ul) were derivatized by adding 90μl of O-methoxyamine hydrochloride solution in anhydrous pyridine and mixed vigorously for 1 min, and incubated at 65 °C for 30 min in a heating block. Subsequently, 14oμl of MSTFA was added and the extracts were incubated at 60 °C for 60 min. Samples were then made up to a volume of 800μl with hexane prior to GC-MS analysis. GC coupled with mass spectrometer (Thermo Scientific, FL, USA) was used to determine the metabolites. TG-5MS fused-silica capillary column (30 m × 250 μm) was utilized to separate the derivatives. The injector temperature was set at 250 °C. Helium, the carrier gas, with constant flow rate of 1.1 ml/min and the column temperature was initially kept at 67 °C for 3min, then ramped to 230 °C at a rate of 5°C/min and then finally increased to 290 °Cat for 20 min. Electron ionization (EI+) mode used for mass detection. The Mass spectra were acquired at the scan ranged between m/z of 45-800. The sample volume of 1 μl was injected for GC-MS analysis [24].

Statistical Analysis

The average larvicidal test mortality data were subjected to log probit analysis for calculating LC50 and LC90, and other statistics at 95% fiducially limits of upper confidence limit and lower confidence limit, and chi-square values were calculated by using the IBM SPSS Statistics 21.0 software. The result with p<0.005 were considered to be statistically significant [25, 26].

Results

Phytochemicals in plants have been widely used as an alternative to synthetic pesticides/insecticides as a part of integrated vector management against mosquitoes. Cost effective, novel plant based insecticides. Phytochemicals and their efficacy against the mosquito larvae have been reviewed extensively. Botanical Name, Tamil Name, parts used and methods used for preparation of plant crude extract used in the present study are given in [Table 1]. Mortality of Anopheles stephensi and Culex quinquefasciatus after the treatment of ethanol extract of Ipomoea carnea(leaf) extract, methanol extract of Commiphora caudate (leaf), methanol extract of Euphorbia antiquorum (latex) extract and acetone extract of Acalpulco sennaalata( leaf) extract were observed. The LC50 and LC90 values after 24hrs of exposure were calculated using probit regression analysis (95% log probit confident limit) with IBM SPSS 21.0 Software. The larval mortality of Anopheles stephensi [late third instar larvae] after the treatment of four different medicinal plants at different concentrations (50 to300) is shown in [Table 2]. The larval mortality of Culex quinquefasciatus (late third instar larvae) after treatment of four different plants at different concentrations (50 to 300 ppm) is given in [Table 3]. 21% mortality was noted in late third instar larvae after treatment of Ipomoea carnea at 50ppm, whereas it has been increased to 100% at 300 ppm of Ipomoea carnea leaf extract treatment. 13% mortality was noted in late third instar larvae after treatment of Commiphora caudate leaf extract at 50ppm, whereas it has been increased to 97% at 300 ppm with Commiphora caudate leaf extract treatment [Table 5]. 14% mortality was noted in late third instar larvae after treatment of Euphorbia antiquorum at 50ppm, whereas it has been increased to 80% at 300 ppm with Euphorbia antiquorum latex extract. 5% mortality was noted in late third instar larvae after treatment of Acalpulco senna-alata at 50ppm, whereas it has been increased to 78% at 300ppm of Acalpulco senna-alata leaf extract treatment. All the values were in four replicates given in [Table 4, 6]. 18% mortality was noted in late third instar larvae after treatment of Ipomoea carnea at 50 ppm, whereas it has been increased to 100% at 300 ppm of Ipomoea carnea leaf extract treatment.15% mortality was noted in late third instar larvae Culex quinquefasciatus after treatment of Commiphora caudate leaf at 50ppm, whereas it has been increased to 100% at 300ppm of Commiphora caudate leaf extract treatment. 11% mortality was noted in late third instar larvae of Culex quinquefasciatus after treatment of Euphorbia antiquorum at 50ppm; whereas it has been increased to 80% at 300 ppm of Euphorbia antiquorum latex extract treatment. 7% mortality was noted in late third instar larvae of Culex quinquefasciatus after treatment of Acalpulco senna-alata at 50 ppm, whereas it has been increased to 78% at 300 ppm of Acalpulco senna-alata leaf extract treatment. All values are in four replicates given from [Table 7-11]. The LC50 and LC90 values for Anopheles stephensi are represented as follows: LC50 value of Ipomoea carnea is 26.970, LC50 value of Commiphora caudate is 47.854, LC50 value of Euphorbia antiquorum is 37.579, and LC50 value of Acalpulco sennaalata is 55.581. LC90 value of Ipomoea carnea is 34.382, LC90 value of Commiphora caudate is 58.761, LC90 value of Euphorbia antiquorum is 48.012, and LC90 value of Acalpulco senna-alata is 69.085. All values are in four replicates in [Table 12]. Likewise, in this study, the larvicidal activity of Culex quinquefasciatus was tested at different concentrations of four different plants mentioned above. The LC50 and LC90 value are represented as follows: LC50 value of Ipomoea carnea is 35.973, LC50 value of Commiphora caudate is 42.775, LC50 value of Euphorbia antiquorum is 40.082, and LC50 value of Acalpulco sennaalata is 52.794. LC90 value of Ipomoea carnea is 44.275, LC90 value of Commiphora caudate is 51.937, LC90 value of Euphorbia antiquorum was 51.501, and LC90 value of Acalpulco senna-alata is 66.284. . Present study showed that highest mortality rate was found in the ethanol leaf extract of Ipomoea carnea (100% mortality) p value is (p≤0.001) followed by methanol leaves extract of Commiphora caudate (97%) p value is (0.008) given in [Table 13]. Chemical components present in Ipomoea carnea plant analysed by Gas Chromatography –Mass Spectrometry is cited in [Table 14]. Chemical components present in Commiphora caudate plant analyzed by Gas Chromatography –Mass Spectrometry is listed in [Table15]. Chemical components present in Euphorbia antiquorum plant analysed by Gas Chromatography –Mass Spectrometry is given in [Table16]. Chemical components present in Acalpulco senna-alata plant analyzed by Gas Chromatography –Mass Spectrometry is listed in [Table 17].

S.No Botanical Name Tamil Name Parts Used Method of Extraction
1 Ipomoea carnea Neyveli kattamanakku Leaves Maceration method
2 Commiphora caudate Kiluvai Leaves Maceration method
3 Euphorbia antiquorum Sadhurakalli Latex Maceration method
4 Acalpulcosenna-alata Vandukadi Leaves Maceration method

Table 1: Botanical Name, Tamil Name, parts used and methods used for preparation of plant crude extract.

S. No Plants name Ethanol Methanol Acetone
1 Ipomoea carnea 100% _ _
2 Commiphora caudate _ 97% _
3 Euphorbia antiquorum _ 86% _
4 Acalpulco senna-alata _ _ 78%

Table 2: Larvicidal activity of different plant extracts in Anopheles stephensi.

S. No Plants name Ethanol Methanol Acetone
1 Ipomoea carnea 100% _ _
2 Commiphora caudate _ 100% _
3 Euphorbia antiquorum _ 80% _
4 Acalpulco senna-alata _ _ 78%

Table 3: Larvicidal activity of different plant extracts in Culex quinquefasciatus.

Doses in
ppm Total no of larvae Mortality (%)
50 100 13%
100 100 20%
150 100 45%
200 100 63%
250 100 78%
300 100 97%
Control 100 0%

Table 4: Larvicidal activity of Commiphora caudate against Anopheles stephensi.

Doses in Mortality
ppm Total no of larvae (%)
50 100 21%
100 100 51%
150 100 67%
200 100 76%
250 100 82%
300 100 100%
Control 100 0%

Table 5: Larvicidal activity of Ipomoea carnea against Anopheles stephensi.

Doses in Mortality
ppm Total no. of larvae (%)
50 100 14%
100 100 28%
150 100 48%
200 100 67%
250 100 77%
300 100 86%
Control 100 0%

Table 6: Larvicidal activity of Euphorbia antiquorum against Anopheles stephensi.

Doses in Mortality
ppm Total no of larvae (%)
50 100 5%
100 100 20%
150 100 35%
200 100 57%
250 100 68%
300 100 78%
Control 100 0%

Table 7: Larvicidal activity of Acalpulco senna-alata against Anopheles stephensi.

Doses in
ppm Total no of larvae Mortality%
50 100 15%
100 100 27%
150 100 59%
200 100 76%
250 100 85%
300 100 100%
Control 100 0%

Table 8: Larvicidal Activity of Commiphora caudate against Culex quinquefasciatus.

Mortality
Doses in ppm Total no of larvae (%)
50 100 18%
100 100 38%
150 100 63%
200 100 75%
250 100 89%
300 100 100%
Control 100 0%

Table 9: Larvicidal activity of Ipomoea carnea against Culex quinquefasciatus.

Mortality
Doses in ppm Total no of larvae (%)
50 100 11%
100 100 27%
150 100 43%
200 100 63%
250 100 74%
300 100 80%
Control 100 0%

Table 10: Larvicidal activity of Euphorbia antiquorum against Culex quinquefasciatus.

Mortality
Doses in ppm Total no of larvae (%)
50 100 7%
100 100 20%
150 100 32%
200 100 56%
250 100 69%
300 100 78%
Control 100 0%

Table 11: Larvicidal activity of Acalpulcosenna-alata against Culex quinquefasciatus.

S.NO Name of the Plant Concentration % LC50 LC90 X2
extracts on in ppm Mortality (LCL±UCL) (LCL±UCL) df=4
1 Ipomoea carnea 50ppm 100ppm 150ppm 200ppm 250ppm 21%
300ppm 51% 26.97 34.382 13.794
67% (7.839±45.615) (11.860±54.472)
76%
82%
100%
2 Commiphora caudate 50ppm 100ppm 150ppm 200ppm 250ppm 13%
300ppm 20%
45% 47.854 58.761
63% (10.498±77.537 (16.385±89.548)
78% 22.244
97%
3 Euphorbia antiquorum 50ppm 100ppm 150ppm 200ppm 250ppm 14%
300ppm 28%
48% 55.581 69.085 5.124
67% (43.371±66.448 (50.308±80.345)
77%
86%
4 Acalpulco senna-alata 50ppm 100ppm 150ppm 200ppm 250ppm 5%
300ppm 20%
35% 37.576 48.012 1.769
57% (27.990±46.655 (37.400±57.768)
68%
78%

Table 12: LC50 Values of plant extract after 24 hrs against Anopheles stephensi.

S. Name of the Plant extracts concentration in ppm % LC50 (LCL±UCL) LC90 (LCL±UCL) X2
No Mortality df=4
1 Ipomoea carnea 50ppm 100ppm 150ppm 200ppm 250ppm 18%
300ppm 38% 35.973 44.275 15.308
63% (13.687±55.532) (19.267±64.850)
75%
89%
100%
2 Commiphora caudate 50ppm 100ppm 15% 21.077
27% 42.775 51.936
150ppm 200ppm 250ppm 59% (14.264±66.242) (20.303±76.333)
300ppm 76%
85%
100%
3 Euphorbia antiquorum 50ppm 100ppm 150ppm 200ppm 250ppm 11%
300ppm 27% 40.082 51.501 2.672
43% (29.675±49.858) (39.998± 61.982)
63%
74%
80%
4 Acalpulcosenna alata 50ppm 100ppm 150ppm 200ppm 250ppm 7%
300ppm 20% 52.794 66.284 5.37
32% (40.909±63.619) (53.590 ±77.567)
56%
69%
78%

Table 13: LC50 Values of plant extract after 24 hrs against Culex quinquefasciatus.

Peak   Value Mol. Weight
S. No Compounds Mol.Formula
1 Hexadecanoic acid, methyl ester 74 C17H34O2 270
2 Pyridine 79 C5H5N 79
3 Benzaldehyde 4 methyl 91 C8H8O 120
4 Levoglucosenone 98 C6H6O3 126
5 Cyclooctasilaxane, hexadecamthyl 355 C10H30O5Si5 370
6 Pentasiloxane, dodecamethyl 281 C12H36O4Si5 384
7 Benzaldehyde 4 methyl, O-methyloxime 91 C9H12NO 149
8 2,4,4,7 Tetramethyl- Octa-5,7- dien-3-one 109 C12H20O 180
9 Cyclohexasiloxane, dodecamethyl 73 C12H36O6Si6 444
10 Acetic acid, trifluro- dodecyl ester 69 C14H25F3O2 282
11 Cycloheptasiloxane, tetradecamethyl 73 C14H42O7Si7 518
12 Phenol 2,4-bis(1,1-dimethylethyl) 191 C14H22O 206
13 3,7,11,15,Tetramethyl 2-hexadecen-1-ol 81 C20H40O 296
14 n-Hexadecanoic acid 43 C16H32O2 256
15 Methyl 9-cis, 11-trans- octadecadienoate 67 C19H34O2 294
16 10-Octadecenoic acid, methyl ester 55 C19H36O2 296
17 Methyl stearate 74 C19H38O2 298
18 cis- Vaccenic acid 55 C18H34O2 282

Table14: List of chemical components present in Ipomoea carnea plant analysed by Gas Chromatography –Mass Spectrometry.

Peak Mol.
S.No. Compounds Value Mol.Formula Weight
1 1,2,4 Butanetriol 106 C4H10O3 75
2 Bensa 4 methyl, O-methyloxime 149 C9H11NO 91
3 Succinimide 99 C4H5NO2 99
4 Benzoic acid 122 C7H6O2 105
5 Benzene 1 methyl 2 (methylthio) 138 C8H10S 138
6 Phenol 2 ethoxy 138 C8H10O2 110
7 Cyclohexasiloxane, dodecamethyl 444 C12H36O6Si6 73
Trimethylsiloxy-6 hexadecenoic acid,
8 emthyester 356 C20H40O3Si 73
9 1H Isoindole-1,3 (2H) dione,2methyl 161 C9H7NO2 161
10 Dichoroacetic acid, tetradecylester 324 C16H30Cl2O2 43
11 2 propenoic acid 3phenyl 148 C9H8O2 147
12 Cycloheptasilaxane,tetra decamethyl 518 C14H42O7Si7 73
13 D-ribose, 2 deoxy-bis (thio hepty)-dithioacetal 380 C19H40O3S2 117
14 Cyclooctasilaxane, hexadecamthyl 592 C16H48O8Si8 355
15 1H Indene,3 butyl 1 methyl 186 C14H18 143
16 n- Hexadecanoic acid 256 C16H32O2 43
17 cis- Vaccenic acid 282 C18H34O2 55
18 Octadecanoic acid 284 C18H36O2 43
19 Pyridine 79 C5H5N 79
20 Cyclopentasiloxane, decamethyl 370 C10H30O5Si5 355
21 1,2, Propanediol 3 chloro 110 C3H7ClO2 43
22 N- methoxy – N methylacetamide 103 C4H9NO2 43

Table 15: List of chemical components present in Commiphora caudate plant analyzed by Gas Chromatography Mass Spectrometry.

Peak Mol.
S.No. Compounds Value Mol.Formula Weight
C5H5N
1 Pyridine 79 79
C8H8O
2 Benzaldehyde 2 methyl 91 120
C10H30O5Si5
3 Cyclopentasiloxane, decamethyl 355 370
C9H11NO
4 Benzaldehyde 4 methyl, O-methyloxime 91 149
C12H36O6Si6
5 Acetic acid, trifluro- dodecyl ester 73 282
C14H42O7Si7
6 Phenol 2,4-bis(1,1-dimethylethyl) 69 206
C14H22O
7 3,7,11,15,Tetramethyl 2-hexadecen-1-ol 191 296
C20H40O
8 n-Hexadecanoic acid 81 256
C18H36O2
9 Hexadecanoic acid, methyl ester 88 284
C18H34O2
10 trans -13-Octadecenoic acid 55 282
C18H34O2
11 E,E,Z-1,3,12-Nonadecatriene-5,14-diol 55 294
C20H3402
12 Ethyl 9,12,15-octadecatrienoate 79 306

Table 16: List of chemical components present in Euphorbia antiquorum plant analysed by Gas Chromatography –Mass Spectrometry.

Peak Mol.
S.No. Compounds Value Mol.Formula Weight
1 Pyridine 52 C5H5N 79
2 Cyclotetrasiloxane, octamethyl 281 C8H24O4Si4 296
3 Silicic acid, diethyl bis(trimethylsilyl) ester 207 C10H28O4Si3 296
4 Benzaldehyde, 4-methyl 91 C8H8O 120
5 Benzene, 1-(chloromethyl)-4-methyl 150 C8H9Cl 140
6 Cyclopentasiloxane, decamethyl 355 C10H13O5Si5 370
7 Pentasiloxane, dodecamethyl 281 C12H36O4Si5 384
8 Benzaldehyde, 2,4-dimethyl- 133 C9H10O 134
9 Tricyclo[4.3.1.1 (3,8)]undecane, 1-methoxy 109 C12H2OO 180
10 Cyclohexasiloxane, dodecamethyl 73 C24H50O4Si2 444
Dodecanedioic acid, bis(tert-butyldimethylsilyl)
11 ester 401 C24H50O4Si2 458
12 Acetic acid, trifluoro-, dodecyl ester 69 C14H2F3O2 282
13 Nonane, 1-chloro 43 C9H19CL 162
14 Cycloheptasiloxane, tetradecamethyl 73 C14H42O7SI7 444
15 Phenol, 2,4-bis(1,1-dimethylethyl) 191 C14H22O 206
16 Oleic acid, eicosyl ester 57 C38H74O2 562
17 n-Hexadecanoic acid 43 C16H32O2 256
18 Cyclodecasiloxane, eicosamethyl 73 C20H60O16SI10 740

Table 17: List of chemical components present in Acalpulco senna-alata plant analyzed by Gas Chromatography –Mass Spectrometry.

Discussion

Plant extracts are recognized as cheap and eco-friendly bio pesticides (68, 69). For a given plant species to be classified as an effective insecticide, the secondary metabolites and plant metabolism are indispensable. Such metabolites can vary with the area, season, method of collection and time of collection etc [27, 28]. In the present study, four different plant extract of Ipomoea carnea, Commiphora caudate, Euphorbia antiquorum, Acalpulco senna-alata were tested against Anopheles stephensi and Culex quinquefasciatus. For example, in line with our observations, a study reported, Ipomoea cairica as an effective larvicidal agent against dengue vector such as Aedes Aegyptus and Aedes albopictus [13]. Further, the authors observed 100% mortality Ipomoea cairica. Further studies using this plant by a different group showed that the highest Mortality was observed in acetone extract of Ipomoea cairica leaf with LC50 of 101.94 and 105.59 and LC90 of 447.78 and 321.56 against dengue vector respectively [11]. Several research groups have characterized, the mosquitocidal activity of different plant extracts, in one such study, repellent activity of leaf Commiphora caudate extract against malarial, dengue, filariasis vector mosquitoes was reported (4). Very recently, highest mortality was found in the ethanol extract of Cadaba indicawith LC50115.70, 96.04, 144.50, 145.75ppm; LC90value of 215.46, 204.98, 233.82, 26.86ppm against dengue vector. Also, Pinus sylvestris and Syzgium aromaticum oils were tested against mosquito and the authors found the LC50 92.56 the LC90 value of 137.80 respectively. Further, the high mortality rate was found in the Syzgium.aromaticum plant. Hexane extract of Citrus sinensis against dengue vector was tested and it showed LC50 value of 446.84mg/L; LC90value of 370.96 mg/L respectively [29]. Not only crude extracts and oils but flowers were also used as insecticidal agents. In one such study the repellent activity of flowering extract of Nerium oleander against filarial vector was tested [10]. In this experiment, hexane flower extract was used and the LC50and LC90 value of 102.54 and 7731.80 were reported. All parts of plants can be used as insecticidal agents, interestingly, in one study it was seen that fruit extract was tested as a larvicidal agent against dengue vector [21]. In the present study, our result showed that similar trend of plants as effective larvicidal agents. We tested the late third instar of Anopheles stephensi and Culex quienquefasciatus by using different concentrations of four different plants. The mortality rate was observed after 24hrs which is followed by LC50 and LC 90 values. Based on the values, it was observed that the plant Ipomoea carnea possess high mortality rate. Based on the plants larvicidal activity, GC-MS was carried out to find the active compounds. It was found that cyclo octasilaxane, hexadecamethyl, cyclopentasiloxane, decamethyl, dodecanedioic acid, bis (tert-butyldimethylsilyl ester are the major chemicals present in the plants. It was also seen that the larvae of Anopheles stephensi were more susceptible to the plant extracts than the larvae of Culex quinquefasciatus. In a nutshell, the results in the present suggest that Ipomoea carnea shows that highest mortality rate and can be used as a larvicidal agent for mosquito vector control programmes.

Conclusion

In the present study larvicidal activity of different plants against Culex quinquefasciatus and Anopheles stephensi were tested. The chosen plants were subjected to crude extraction. Different parts of the plants were used for this purpose and the extraction was done by using methanol (100%), ethanol (100%), and, acetone (70%). Ipomoea carnea leafs were subjected to ethanol extraction. Latex of Euphorbia antiquorum and leaves of Commiphora caudate were subjected to methanol extraction, and Acalpulco senna-alata were subjected to acetone extraction. After 24hrs of exposure plants crude extracts displayed high mortality rates. Amongst the plants tested, highest mortality rate was found in the ethanol extract of Ipomoea carnea and lowest mortality rate was found in acetone extract of Acalpulco sennaalata. In order to find the active components in the plant crude extract, GC-MS was carried out and different compounds were reported. The major active component in Ipomea cornea based on the retention time is cyclooctasilaxane, hexadecamethyl. This plant could be utilized for developing a cost effective and environment friendly new type of larvicidal for mosquito control. This plant can be used as a good alternative for synthetic pesticides. The results reported here open the possibility of further investigations of field trails.

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Citation: Rajalakshmi, Krishnan J (2023) Investigation of Plants Extracts as Potential Larvicidal Agents against Culex quinquefasciatus and Anopheles stephensi. J Bioremediat Biodegrad, 14: 548.

Copyright: © 2023 Rajalakshmi, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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