Larvicidal Activity of Some Plants Extracts and Their Partitioned Fractions Against Culex Quinquefasciatus

International Journal of TROPICAL DISEASE & Health

41(11): 23-34, 2020; Article no.IJTDH.59818 ISSN: 2278–1005, NLM ID: 101632866

Larvicidal Activity of Some Plants Extracts and Their Partitioned Fractions against Culex quinquefasciatus

Funmilayo G. Famuyiwa1*, Francis B. Adewoyin2, Oluyemi J. Oladiran1 and Oluwatosin R. Obagbemi1

1Department of Pharmacognosy, Faculty of Pharmacy, Obafemi Awolowo University, Ile-Ife, Nigeria. 2Drug Research and Production Unit, Faculty of Pharmacy, Obafemi Awolowo University, Ile-Ife, Nigeria.

Authors’ contributions

This work was carried out in collaboration among all authors. Author FGF designed the study and performed the statistical analysis. Authors FGF and FBA wrote the first draft of the manuscript. Authors OJO and ORO carried out the larvicidal assay under the supervision of author FBA. Author FGF managed the literature searches. All authors read and approved the final manuscript.

Article Information

DOI: 10.9734/IJTDH/2020/v41i1130332 Editor(s): (1) Dr. Arthur V. M. Kwena, Moi University, Kenya. Reviewers: (1) Stan Florin Gheorghe, University of Agricultural Sciences and Veterinary Medicine of Cluj-Napoca, Romania. (2) Kishore Yadav Jothula, All India Institute of Medical Sciences (AIIMS), India. Complete Peer review History: http://www.sdiarticle4.com/review-history/59818

Received 04 June 2020 Accepted 10 August 2020 Original Research Article Published 24 August 2020

ABSTRACT

Aim: The methanol extracts of fifteen plants and their partitioned fractions were screened for larvicidal activity against the fourth instar of larvae Culex quinquefasciatus, the vector of lymphatic filariasis with a view to identifying the active ones. Methodology: The plant parts were collected, separately dried and milled. Each powdered material was extracted in methanol at room temperature for 3 days, with agitation. The extract was filtered and concentrated in vacuo. Each extract was tested against the fourth instar larvae of Cx. quinquefasciatus. The methanol extracts were suspended in water and successively partitioned into n-hexane and ethylacetate. Each partitioned fraction was also tested against the fourth instar larvae of Cx. quinquefasciatus. Results: About fifty six percent (56.3%) of the tested extracts had moderate larvicidal activity after 48 hours. The fruit extract of Thevetia neriifolia and the leaf extracts of Calotropis procera and ______

*Corresponding author: Email: [email protected];

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Solanum macrocarpon were the most active. After partitioning the methanol extracts, each of the plant extracts had one or two highly active partitioned fractions after 48 hours. The n-hexane fractions of S. macrocarpon (0.78 ± 0.03 mg/mL) and Spondias mombin (0.81 ± 0.03 mg/mL) were the most active. Conclusion: The non-polar fractions of S. macrocarpon and S. mombin were the most active. Purification of these highly active fractions could lead to the isolation of potent larvicidal compounds that could be used in the control of Cx. quinquefasciatus mosquito.

Keywords: Larvicidal activity; filariasis; partitioned fraction; extracts; Culex quinquefasciatus.

1. INTRODUCTION Pharmacy and voucher specimens were deposited at the Faculty of Pharmacy Herbarium. Lymphatic filariasis is a mosquito-borne parasitic disease that is endemic in many tropical and 2.2 Preparation of the Extracts and subtropical countries. This disease, caused by Fractions thread-like parasitic Wuchereria bancrofti, Brugia malayi or B. timori is transmitted by Culex The leaves were air-dried while the stem bark, quinquefasciatus and certain Anopheles species, fruit and roots were oven-dried at 35ºC and then and it is one of the neglected tropical diseases milled into fine powder using the Warring [1,2]. Controlling the vectors will go a long way in blender. The powdered plant parts were the control of the disease. The control of separately weighed and soaked in methanol for mosquito at the larva stage is efficient as they 72 hours with regular shaking. The extracts were are relatively immobile and a large population filtered and concentrated in vacuo at 35ºC and can be killed in their breeding sites with little this process was repeated twice. Each of the effort [3]. There is an increase in the extracts was suspended in water, successively development of resistance to currently available partitioned with n-hexane and ethylacetate and synthetic larvicides especially in the tropics [4]. concentrated in vacuo to get their corresponding The advantages of larvicides of plant origin over fractions. the synthetic ones cannot be overemphasised, 2.3 Evaluation of the Larvicidal Activity and this has stimulated intensified efforts towards developing plant based larvicides. Search for Stock solution of each of the extracts (25 mg/mL) eco-friendly, safe, low cost and potent plant- and fractions (5 mg/mL) was prepared by based larvicides for the control of mosquitoes solubilising in Tween 80 and diluting with distilled need the preliminary screening of plants to water. They were serially diluted to obtain 25 mL evaluate their larvicidal potentials [5]. Plants that of different concentrations of the test agents, (0, are used folklorically as fish poisons and in the 1, 2, 3, 4, 5 mg/mL for extracts and 0, 0.5, 1.0, treatment of malaria and fever, as well as those 1.5, 2.0 and 2.5 mg/mL for fractions). Twenty-five with reported insecticidal and insect repellent larvae were introduced into each cup and each activities have been suggested as lead in the concentration was replicated five times. The choice of plants to be screened for larvicidal methanol extract of Nicotiana tabacum leaf, a activity [6]. Therefore, in this study, the methanol known insecticidal plant was used as the positive extract of fifteen plants possessing one or more control, and its toxicity was evaluated at 0.2 – 1.0 of these characteristics and their partitioned mg/mL while Tween 80 in distilled water was fractions were screened for larvicidal activity used as the negative control. The number of against Cx. quinquefasciatus. surviving larvae was counted after 24 and 48hours of exposure during which no nutritional 2. MATERIALS AND METHODS supplement was added [7]. The percentage mortalities were calculated and the LC50 and 2.1 Plants Collection LC90 values were predicted using Microsoft Excel program 2010. The larvicidal activity of the Each of the plants listed in Table 1 was collected extracts and their fractions were compared with at various locations in Obafemi Awolowo that of the positive control and with each other by University, Ile-Ife, Osun State, Nigeria. They subjecting the values to statistical analysis using were identified by Mr. Ogunlowo of the ANOVA and Student Newmann Keul’s post-hoc Department of Pharmacognosy, Faculty of test. P < 0.05 was considered as significant.

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Table 1. List of plants used and their related activities

Name of plants Family Part used Related activity References 1 Anthocleista vogelli Gentianaceae Bark Fever; malaria Anyawu et al., 2018 [8] 2 Calotropis procera Asclepiadaceae Leaf Fever, malaria Bairagi et al., 2018 [9]. 3 Cassia sieberiana Fabaceae Root Fever, malaria, termite resistant Archer et al., 2019 [10]. 4 Delonix regia Fabaceae Bark Fever, malaria Ahmed et al., 2009 [11] 5 Ficus exasperata Moraceae Leaf Fever, malaria, insecticidal Ahmed et al., 2012 [12] 6 Ficus sur Moraceae Leaf Arrow poison, malaria Maroyi, 2013 [13] 7 Ficus vogelli Moraceae Leaf Malaria Igile et al., 2015 [14] 8 Hilleria latifolia Petiveriaceae Leaf Fever, malaria Johnson et al., 2018 [15] 9 Momordica charantia Cucurbitaceae Leaf Fever, malaria Pani et al., 2015 [16] 10 Psidium guajava Asclepiadaceae Leaf Fever Naseer et al., 2018 [17] 11 Senna alata Fabaceae Leaf Insecticidal Adelowo and Oladeji, 2017 [18] 12 Solanum macrocarpon Solanaceae Leaf Insect resistant Kumar and Sadashiva, 1996 [19] 13 Spondias mombin Anacardiaceae Bark Fever, malaria Mitchell and Daly, 2015 [20] 14 Thevetia nerifolia Apocynaceae Bark and Fruit Insecticidal Rajbhar and Kumar, 2014 [21] 15 Vitex doniana Verbanaceae Bark Fever, insect resistant Ibrahim et al., 2013 [22]

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Table 2. Larvicidal activities of the extracts and partitioned fractions against C. quinquefasciatus at 24 hours

S/N Name of plant Methanol extract N-hexane fraction Ethylacetate fraction Aqueous fraction LC50 LC90 LC50 LC90 LC50 LC90 LC50 LC90 1 Anthocleista vogelli 3.92 ± 0.05b 5.68± 0.03 1.82 ± 0.04a 3.13 ± 0.08 1.85 ±0.08a 3.46 ± 0.17 6.91 ± 0.78c 12.02 ± 1.37 2 Calotropis procera 3.83 ± 0.15b 5.91 ± 0.07 2.05 ± 0.16 a 3.74 ± 0.36 2.13±0.09 a 3.92 ± 0.16 7.60 ± 0.71c 13.00 ± 1.31 3 Cassia sieberiana 6.28 ± 0.73 c 9.95 ± 1.36 2.96 ± 0.24b 5.25 ± 0.50 2.23±0.05 a 4.06 ± 0.13 6.34 ± 0.71c 11.15 ± 1.36 4. Delonix regia 6.35 ± 0.49 b 10.78 ± 0.91 3.06 ± 0.22 a 5.20 ± 0.45 3.57±0.20 a 6.15 ± 0.42 7.10 ± 0.57 b 12.50 ± 0.97 5. Ficus exasperata 5.04 ± 0.41c 8.92 ± 0.76 2.66 ± 0.03 a 4.91 ± 0.06 3.63 ±0.03b 6.33 ± 0.04 4.59 ± 0.67 c 7.74 ± 1.24 6. Ficus sur 12.23 ±0.91c 21.05 ± 0.50 6.52 ± 0.53 b 11.4 ± 1.02 3.32 ±0.43a 5.70 ± 0.87 3.26 ± 0.17 a 5.81 ± 0.33 7. Ficus vogelli 3.79 ± 0.25b 6.23 ± 0.56 1.45 ± 0.10 a 2.83 ± 0.17 6.06 ±0.76c 10.91±1.32 8.38 ± 0.13 d 14.38 ± 0.13 8. Hilleria latifolia 8.29 ± 1.08 c 13.84 ± 2.02 3.62 ± 0.59 a 6.28 ± 0.14 3.81 ±0.25a 7.01 ± 0.44 5.42 ± 0.84 b 9.11 ± 1.67 9. Momordica charantia 5.01 ± 0.13c 7.81 ± 0.12 4.05 ± 0.19 a 7.18 ± 0.33 4.81 ±0.34b 8.44 ± 0.73 4.75 ± 0.03b 8.50 ± 0.02 10. Psidium guajava 4.52 ± 0.13b 7.86 ± 0.04 3.96 ± 0.74 a 6.63 ± 1.29 6.81 ±1.44 c 11.6 ± 2.65 8.50 ± 0.00 d 14.5 ± 0.00 11. Senna alata 5.23 ± 0.11c 9.28 ± 0.03 1.61 ± 0.15 a 2.98 ± 0.27 1.90 ±0.15b 3.51 ± 0.25 8.45 ± 0.05 d 14.45 ± 0.05 12. Solanum macrocarpon 3.34 ± 0.09b 5.46 ± 0.24 0.93 ± 0.11 a 1.83 ± 0.19 1.20 ±0.05 a 2.31 ± 0.11 7.58 ± 0.60 c 13.13 ± 1.05 13. Spondias mombin 7.90 ± 1.16 c 13.48 ± 2.20 0.83 ± 0.02 a 1.65 ± 0.05 6.97 ±0.58 c 12.12±0.94 4.15 ± 0.67 b 7.20 ± 1.27 14. Thevetia nerifolia bark 4.75 ± 0.11b 7.83 ± 0.09 1.60 ± 0.05 a 3.04 ± 0.10 1.51 ±0.03a 2.89 ± 0.06 4.96 ± 0.72 b 8.60 ± 1.37 15. Thevetia nerifolia fruit 3.64 ± 0.12c 5.84 ± 0.09 1.64 ± 0.10 b 2.69 ± 0.19 1.09 ±0.04a 2.19 ± 0.07 4.00 ± 0.27 c 7.04 ± 0.47 16. Vitex doniana 10.81 ±0.36c 18.23 ± 0.25 1.27 ± 0.07 a 2.51 ± 0.11 4.34 ±0.33b 7.62 ± 0.58 4.75 ± 1.00 b 8.04 ± 1.84 17. Nicotiana tabacum LC50 1.94 ± 0.25 mg/mL LC90 3.26 ±0 .46 mg/mL Key: LC50 and LC90: Concentration in mg/mL that killed 50 and 90% respectively; a-d: LC50 values with different superscripts across the rows are significantly different (P=0.05, one-way analysis of variance followed by the Student–Newman–Keuls’ test)

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Table 3. Larvicidal activities of the extracts and partitioned fractions against C. quinquefasciatus at 48 hours

S/N Name of plant Methanol extract N-hexane fraction Ethylacetate fraction Aqueous fraction LC50 LC90 LC50 LC90 LC50 LC90 LC50 LC90 1. Anthocleista vogelli 2.84 ±0.13 c 5.31 ± 0.01 1.20 ± 0.02 a 2.33 ± 0.04 1.46 ± 0.05 b 2.79 ± 0.09 2.81 ± 0.15 c 5.21 ± 0.27 2. Calotropis procera 2.06 ± 0.09 b 4.26 ± 0.06 1.30 ± 0.04 a 2.45 ± 0.06 1.20 ± 0.05 a 2.30 ± 0.11 6.31 ± 1.10 c 7.19 ± 1.46 3. Cassia sieberiana 6.36 ± 0.49 c 11.62 ± 0.87 1.90 ± 0.10 a 3.31 ± 0.21 1.77 ± 0.10 a 3.36 ± 0.17 2.13 ± 0.17 b 3.94 ± 0.31 4. Delonix regia 5.09 ± 0.70 d 8.88 ± 1.35 1.72 ± 0.11 a 3.02 ± 0.18 2.18 ± 0.13 b 3.89 ± 0.20 3.56 ± 0.26 c 5.46 ± 0.51 5. Ficus exasperata 3.65 ± 0.16 c 6.42 ± 0.28 1.70 ± 0.12 a 3.26 ± 0.22 2.58 ± 0.02 b 4.67 ± 0.03 3.45 ± 0.31 c 6.07 ± 0.63 6. Ficus sur 10.17 ± 0.25 c 16.83± 0.30 4.03 ± 0.14 b 7.13 ± 0.29 1.73 ± 0.10 a 3.02 ± 0.18 1.52 ± 0.11 a 2.89 ± 0.21 7. Ficus vogelli 3.37 ± 0.13 c 5.45 ± 0.35 1.32 ± 0.11 a 2.59 ± 0.21 2.56 ± 0.14 b 4.62 ± 0.24 5.42 ± 1.56 d 9.49 ± 2.27 8. Hilleria latifolia 6.70 ± 0.97 b 12.08 ± 1.80 2.33 ± 0.16 a 4.17 ± 0.32 2.03 ± 0.19 a 3.64 ± 0.36 5.39 ± 0.91 b 9.31 ± 1.69 9. Momordica charantia 4.61 ± 0.15 c 7.71 ± 0.03 1.53 ± 0.04 a 2.96 ± 0.08 2.45 ± 0.03 b 4.56 ± 0.04 4.75 ± 0.03 c 8.50 ± 0.02 10. Psidium guajava 3.85± 0.35 b 7.27 ± 0.14 2.39 ± 0.25 a 4.33 ± 0.45 4.27 ± 0.44 b 7.54 ± 0.74 4.83 ± 0.21 c 8.67 ± 0.41 11. Senna alata 4.16 ± 0.28 b 7.43 ± 0.05 1.34 ± 0.15 a 2.60 ± 0.25 1.49 ± 0.08 a 2.81 ± 0.14 6.59 ± 0 61 c 11.53 ± 1.09 12. Solanum macrocarpon 2.35 ± 0.05 c 4.22 ± 0.13 0.78 ± 0.03 a 1.55 ± 0.07 0.91 ± 0.02 b 1.84 ± 0.02 7.58 ± 0.60 d 13.13 ± 1.05 13. Spondias mombin 5.85 ± 0.70 c 10.04 ± 1.25 0.81 ± 0.03 a 1.51 ± 0.07 4.48 ± 0.42 c 8.06 ± 0.75 2.68 ± 0.18 b 4.76 ± 0.38 14. Thevetia nerifolia bark 3.66 ± 0.12 b 6.02 ± 0.02 1.22 ± 0.06 a 2.30 ± 0.09 1.13 ± 0.05 a 2.23 ± 0.08 3.49 ± 0.49 b 6.14 ± 0.97 15. Thevetia nerifolia fruit 2.85 ± 0.04 b 5.01 ± 0.02 1.01 ± 0.05 a 2.04 ± 0.08 0.85 ± 0.03 a 1.76 ± 0.05 3.20 ± 0.32 b 5.70 ± 0.58 16. Vitex doniana 7.54 ± 0.61 c 12.93 ± 1.24 0.78 ± 0.01 a 1.55 ± 0.01 2.22 ± 0.10 b 4.00 ± 0.18 2.29 ± 0.16 b 3.75 ± 0.34 17 Nicotiana tabacum LC 50 1.51 ± 0.18 mg/mL LC90 2.52 ± 0.40 mg/mL Key: LC50 and LC90: Concentration in mg/mL that killed 50 and 90% respectively; a-d: LC50 values with different superscripts across the rows are significantly different (P=0.05, one-way analysis of variance followed by the Student–Newman–Keuls’ test)

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3. RESULTS AND DISCUSSION extract of any of its morphological parts in literature. 3.1 Results Calotropis procera leaf: The moderate activity of The results of the activities of the methanol the methanol extract was in line with previous extracts and the partitioned fractions are as reports on the aqueous leaf extract having presented in Tables 2 and 3. remarkable larvicidal, adult emergence inhibitor, repellent and oviposition deterrent effects against 3.2 Discussion mosquito species and Musca domestica [29-31]. The n-hexane fraction and ethylacetate fraction Diseases such as filariasis, dengue fever, yellow had better activity than the aqueous fraction and fever, malaria, Japanese encephalitis and the extract throughout the test period. chikungunya transmitted by mosquitoes are still major health problems in most countries [23]. Cassia sieberiana root DC: The root extract was Even though chemical control program has been inactive till 48hours despite the significant (p< carried out for a long time, these mosquitoes 0.05) inhibition of the hatching of the eggs of remain because of the development of resistance Haemonchus contortus, a gastrointestinal to the available synthetic products [24]. This has parasite by its leaf and root extracts [32]. necessitated the continued search and Larvicidal activity had been reported for the leaf development of environmentally safe, extract of Cassia fistula and the seed extract of biodegradable, and low cost chemical Cassia tora and Cassia occidentalis against Cx. compounds from plants which can be used by Quinquefasciatus [33-35] but there was no individuals and communities in specific previous report for C. sieberiana. After situations. There is therefore a provocative partitioning, all the resulting fractions had better interest in research for larvicidal compounds from activity than the extract. The n-hexane and natural sources [25]. In this study the methanol ethylacetate fractions were comparably more extract of fifteen plants were screened. At 24 active than the aqueous fraction. hours, five of the plant extracts namely A. vogelli, C. procera, F. vogelli, S. macrocarpon and T. Delonix regia (Boj.exHook.) Raf. Bark: The low neriifolia fruit (31.25%) had moderate larvicidal activity of the methanol extract of the bark in this activity (2.0 < LC50< 4.2 mg/mL) while the study is similar to the low mortality reported for remaining eleven extracts (68.75%) had low the ethanol and aqueous extracts of the seed activity with LC50>4.2 mg/mL [6,26]. After 48 against the larvae of An. gambiae [36]. The high hours of exposure to these test agents, the activity of the various solvent extracts and the activities of F. exasperata, P. guajaya, S. alata essential oil of the flower against mosquitoes, and T. neriifolia bark improved making the leaf-eating caterpillars and beetles like Hyblaea moderately active ones to be 56.3% (Table 3). It puera, Sitophilus zeamais, Calosobruchus had been earlier suggested that plant extracts maculatus and Pericallia ricini had been reported which have some degrees (moderate or low) of [37-39]. This may imply that the larvicidal activity should be fractionated and tested before compound(s) of this plant is concentrated in the discarding them [27]. Therefore, these methanol flower. The n-hexane and ethylacetate extracts were suspended in water, successively partitioned fractions of the bark extract used in solvent partitioned and tested against Cx. this study had high larvicidal activities while the quinquefasciatus larvae. aqueous fraction was inactive. The activity of the aqueous fraction greatly improved at 48 hours. Anthocleista vogelli Planch bark: The methanol extract of A. vogelli bark, commonly called Ficus exasperata Vahl, Ficus sur Forssk and cabbage tree, was moderately active against the Ficus vogelli Miq. Leaves: The methanol extract test organism. The essential oils from the leaf of F. vogelli was the most active, followed by and stem bark obtained by hydrodistillation was F.exasperata while F. sur was the least active highly toxic to the third instars larvae of Culex throughout the test period. High activity had been mosquito [28]. The n-hexane and the reported for the aqueous and ethanol extracts ethylacetate fractions had comparable activities and the powder of F. exasperata leaf against at 24 hours while the aqueous fraction was snails, C. maculatus and S. zeamais [12]. The inactive. There was a significant improvement in inactivity of F. sur methanol extractin this study the activity of the n-hexane fraction at 48 hours was contrary to an earlier report of high activity making it the most active fraction. There was no [40]. There is no previous report of the larvicidal previous report of the larvicidal activity of the activity of F. vogelli against any vector. The

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methanol extract of Ficus benghalensis, a related slightly at 48 hours. The observed activity was species, showed good larvicidal activity against similar to earlier reports against Ae. aegypti Cx. quinquefasciatus [23] while gluanol acetate, mosquito [48]. However, the essential oil and its potent against fourth instar larvae of Ae. aegypti, major compounds 1, 8-cineole and carvacrol An. stephensi and Cx. quinquefasciatus was exhibited significant larvicidal activity against Ae. isolated from the moderately active acetone aegypti, Trogoderma granarium and Chaoborus extract of F. racemosa bark [41]. The n-hexane plumicornis [49]. There was no significant fraction of F. vogelli and F.exasperata were the improvement in the activity of the resulting most active while for F. sur, the aqueous fraction partitioned fractions; the most active n-hexane was the most active. fraction had comparable activity with the extract while the others were worse off. At 48 hours, n- Hilleria latifolia (Lam.) H. Walter leaf: The extract hexane fraction was the most active while the had low activity against the test organism ethylacetate and aqueous fractions maintained throughout the test period (Tables 2 and 3). This their low activity. is the first report of the larvicidal activity of this plant extract against Cx. quinquefasciatus larvae. Senna alata L. (Roxb) leaf: The S. alata Partitioning the extract gave a moderately active (synonym Cassia alata) leaf extract had low n-hexane and ethylacetate fractions at 24 hours activity throughout the test period. This differed with improvement after 48 hours of exposure. from the high activity reported against An. Their activities were significantly less than that of gambiae, Cx. quinquefasciatus and Ae. aegypti N. tabacum, the positive control used. However, [50-52]. Moderate activities had been reported their purification can lead to the isolation of for some other species of Cassia like Cassia potent larvicidal compounds. For example, the n- arereh stem bark methanol extract against C. hexane fraction of Blighia sapida extract was not quinquefasciatus, the seed petroleum ether as active as Endosulphan (a commercial extract of C. siamea leaf aqueous extract, S. insecticide) but its purification led to the isolation auriculata, S. tora and C. fistula against An. of friedelin and α-amyrin with high activities stephensi and Ae. aegypti, ethanol extract of C. against Ae. aegyti [42]. occidentalis leaf [53-55]. Partitioning the extract gave a highly active n-hexane and ethylacetate Momordica charantia L. leaf: The low activity of fractions. Similarly, the hexane extract of the fruit M. charantia leaf extract in this study is contrary caused high lethality to Callosobruchus chinensis to the high activity reported against Ae. aegypti [56]. larvae [43]. This could be due to the difference in the mosquito species and the method of Solanum macrocarpon L. leaf: The extract was extraction; in that report; Soxhlet method was moderately active against the test organism. used while in this study, maceration was used. There had been reports of the larvicidal activity of Also, the hexane extract and the nanopowder of Solanum nigrum leaf and berry extract and the the fruit had higher activity than the methanol phytosteroid isolated from the leaf against Ae. extract used in this study against Cx. aegypti, Cx. vishnui and An. subpictus [57,58]. quinquefasciatus and Cx. pipiens [44]. The The potential of the methanol extract and activity of all the partitioned fractions was not fractions of the green fruits of Solanum significantly better than that of the extract at 24 lycocarpum against Cx. quinquefasciatus, hours. At 48 hours, the high activity of the n- aqueous extract of S. tuberosum tuber against hexane fraction is in line with the 100% mortality Cx. quinquefasciatus and An. stephensi had also reported for the hexane extract [45]. Non-polar been previously reported [59,60] but there is no characteristics of larvicidal components of the previous report on this species. The n-hexane fruit extract against fourth instars larvae of An. fraction was highly active at both 24 and 48 stephensi, Cx. quinquefasciatus and Ae. aegypti hours followed by the ethylacetate fraction while had been demonstrated [46]. Momordicine II and the aqueous fraction was inactive. The high IV isolated from the butanol fraction of this plant activity of the n-hexane fraction in the study was deterred oviposition of Liriomyza trifolii (a major similar to the high activity reported for the leaf miner pest of a wide variety of vegetables, petroleum ether extract of Solanum trilobatum floricultures, and ornamental plants) significantly [61]. [47]. Spondias mombin L. bark: Despite the low Psidium guajava L. leaf: The low activity of the activity demonstrated by S. mombin bark extract, leaf methanol extract at 24 hours improved its partitioning resulted into a highly active n-

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hexane fraction. This was the trend of activity for purification of each of these highly active the leaf methanol extract and its n-hexane fractions could lead to the isolation of potent fraction against Cx. quinquefasciatus and Ae. larvicidal compounds that could be used in the aegypti [62]. However dichloromethane fraction control of Cx. quinquefasciatus mosquito. was the most active in the adulticidal and repellency assays against adult Ae. aegypti CONSENT mosquito [63,64]. The aqueous fraction was moderately active at 48 hours while the It is not applicable. ethylacetate fraction had comparable activity to the extract throughout the test period. ETHICAL APPROVAL

Thevetia nerifolia L. fruit and bark: The extract of T. nerifolia (syn. Thevetia peruviana) fruit and It is not applicable. bark had moderate and low activities respectively at 24 hours. Contrarily, its various leaf extracts, ACKNOWLEDGEMENT gave promising effects on the larvae of Culex, An. stephensi and Ae. aegypti mosquito and M. The authors appreciate the assistance of Mr. O.I. domestica in previous reports [65-67]. Also, the Ogunlowo of the Department of Pharmacognosy, methanol extract of the stem reportedly had Obafemi Awolowo University, Ile-Ife, Nigeria antifeedant and stomach poison activities [68] during plants collection. while the latex caused significant reduction in larval weight and very high larval mortality COMPETING INTERESTS against Spodoptera litura [69]. Partitioning the extracts of both morphological parts led to Authors have declared that no competing hexane and ethylacetate fractions with high interests exist. activities. The hexane and ethylacetate fractions of the bark had comparable activity at both hours REFERENCES while the ethylacetate fraction of the fruit was the most active. 1. Fenwick A. The global burden of neglected tropical diseases. Public Health. Vitex doniana L. bark: The very low activity 2012;126(3):233-236. observed for the bark methanol extract at both 24 DOI: 10.1016/j.puhe.2011.11.015 and 48 hours is similar to the report of the Epub 2012 Feb 9 ethanol extract of the leaf against Anopheles 2. Mitra AK, Mawson AR. Neglected tropical mosquito [70]. High larvicidal activity was diseases: Epidemiology and global reported for the essential oil and leaf methanol burden. Trop. Med. Infect. Dis. 2017;2:36- extract of Vitex negundo and Vitex trifolia against 51. Cx. quinquefasciatus and Ae. aegyti [71-73]. DOI: 10.3390/tropicalmed2030036 Throughout this study period, the n-hexane 3. Ali NOM, El-Rabaa FMA. Larvicidal activity partitioned fraction was highly active while the of some plant extracts to larvae of the ethylacetate fraction was moderately active. mosquito Culex quinquefasciatus (Say). Eur. Rev. Med. Pharmaco. 2010;14:925- 4. CONCLUSION 933. 4. Lima MAA, Oliveira FFM, Gomes GA, About fifty six (56.3%) percent of the tested Lavor PL, Santiago GMP, Nagao-Dias AT, plants had moderate activity against Cx. Arriaga ÂMC, Lemos TLG, Geraldo de quinquefasciatus larvae while the remaining ones Carvalho M. Evaluation of larvicidal activity had low activity. This is the first report of the of the essential oils of plants species from larvicidal activity of the methanol extract of A. Brazil against Aedes aegypti (Diptera: vogelli, C. sieberiana, F. vogelli, H. latifolia and Culicidae). Afr. J. Biotech. 2011;10(55): S. macrocarpon. Each of the moderate and low 11716-11720. acting extracts had one or two highly active DOI: 10.5897/ajb11.1102 fractions after 48 hours supporting the 5. Shivakumar MS, Srinivasan R, Natarajan suggestion of fractionating plant extracts with D. Larvicidal potential of some Indian some degrees (moderate or low) of activity. The medicinal plant extracts against Aedes compounds responsible for larvicidal activity of aegypti (L.). Asian J. Pharm. Clin. Res. these plants have not been identified; therefore 2013;6(3):77-80. ISSN - 0974-2441.

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6. Adebajo AC, Famuyiwa FG, John JD, Idem 16. Pani P, Nahak G, Sahu RK. Review on ES, Adeoye AO. Activities of some larvicidal activity of medicinal plants for Nigerian medicinal plants against Aedes malaria vector control. Int. J. Curr. aegypti. Chin. Med. 2012;3:151-156. Pharmaceut. Rev. Res. 2015;6(2):94-114. DOI: 10.4236/cm.2012.33024 ISSN: 0976-822X. 7. Guidelines for Laboratory and Field 17. Naseer S, Hussain S, Naeem N, Pervaiz Testing of Mosquito Larvicides. M, Rahman M. The phytochemistry and Available:http://whqlibdoc.who.int/hq/2005/ medicinal value of Psidium guajava WHO_CDS-WHOES-GCD- (guava). Clin. Phytoscience. 2018;4:32. 2005.13.df?ua=1 DOI: 10.1186/s40816-018-0093-8 (Accessed on 19 June 2014) 18. Adelowo F, Oladeji O. An overview of the 8. Anyanwu GO, Onyeneke CE, Ofoha PC, phytochemical analysis of bioactive Rauf K, ur-Rehman NR, Usunobun U. compounds in Senna alata. Advances in Diuretic, antidiuretic and laxative activities Biochemistry. 2017;5(5):102-109. of Anthocleista vogelii extracts in rats. Afr DOI: 10.11648/j.ab.20170505.14 J. Tradit Complement Altern Med. 19. Kumar NKK, Sadashiva AT. Solanum 2018;15(1):66-73. macrocarpon: A wild species of brinjal Available:https://doi.org/10.21010/ajtcam.v resistant to brinjal shoot and fruit borer, 15i1.7 Leucinodes orbonalis (Guen.). Insect 9. Bairagi SM, Ghule P, Gilhotra R. Environ. 1996;2:241-242. Pharmacology of natural products: A 20. Mitchell JD, Daly DC. A revision of recent approach on Calotropis gigantea Spondias L. (Anacardiaceae) in the and Calotropis procera. ARS Neotropics. Phyto Keys. 2015;55:38. Pharmaceutical. 2018;59(1):37-44. DOI: 10.3897/phytokeys.55.8489 Available:http://dx.doi.org/10.4321/S2340- 21. Rajbhar N, Kumar A. Pharmacological 98942018000100004 importance of Thevetia peruviana. IJPCS. 10. Archer MA, Agyei TA, Mintah SO, Atta- 2014;3(1):260-263. ISSN: 22775005. Adjei P, Kumadoh D, Asiedu-Larbi J. 22. Ibrahim JA, Egharevba HO, Obokayi OD, Medicinal uses of Cassia sieberiana; A Kunle OF. Some medicinal plants of the review. Int. J. Sci.: Basic Appl. 2019;48(2): family Verbenaceae commonly used in 161-180. Jos, Nigeria - A review. Ewemen J. 11. Ahmed J, Nirmal SA, Rub RA, Budhavale Folklore Med. 2015;1(1):23-30. SK, Pattan SR. An overview of Delonix Available:http://ewemen.com/category/ejfm regia: Chemistry and pharmacological / profile. Pharmacologyonline. 2009;2:544- 23. Govindarajan M. Larvicidal efficacy of 549. Ficus benghalensis L. plant leaf extracts 12. Ahmed F, Mueen AKK, Abedin MZ, Karim against Culex quinquefasciatus Say, AA. Traditional uses and pharmacological Aedes aegypti L. and Anopheles stephensi potential of Ficus exasperata Vahl. Sys L. (Diptera: Culicidae). Eur Rev Med Rev Pharm. 2012;3(1):15-23. Pharmacol. Sci. 2010;14:107-111. 13. Maroyi A. Traditional use of medicinal 24. Das NG, Goswami D, Rabha B. plants in South-Central Zimbabwe: Review Preliminary evaluation of mosquito and perspectives. J. Ethnobiol. Ethnomed. larvicidal efficacy of plant extracts. J. Vect. 2013;9:31-49. Borne Dis. 2007;44:145–148. Available:http://www.ethnobiomed.com/con 25. Prabakar K, Jebanesan A. Larvicidal tent/9/1/31 efficacy of Cucurbitacious plant leaf 14. Igile GO, Utin IC, Iwara IA, Mgbeje BIA, extracts against Culex quinquefasciatus Ebong PE. Ethanolic extract of Ficus (Say). Bioresour. 2004;95(1):113-114. vogelii ameliorates dyslipidemia in diabetic 26. Adebajo AC, Famuyiwa FG, Aliyu albino wistar rats. Int. J. Curr. Res. Biosci. FA. Properties for sourcing Nigerian Plant Biol. 2015;2(8):87-96. larvicidal plants. Molecules. 2014;19:8363- 15. Johnson IS, Ettebong EO, Okokon JE, 8372. Johnson ES. Antipyretic potentials of DOI: 10.3390/molecules19068363 ethanolic extract of Hilleria latifolia leaves 27. Wilcox M, Bodeker G, Rasonaivo P. in albino wistar rats. Int J Herbal Med. Traditional medicinal plants and malaria. 2018;6(6):50-53. E-ISSN: 2321-2187. CRC Press, USA. 2004;4(16):262.

Famuyiwa et al.; IJTDH, 41(11): 23-34, 2020; Article no.IJTDH.59818

28. Aboaba SA, Flamini G, Ibrahim K. Volatile 37. Baskaran V, Narayanasamy P. Traditional constituents and larvicidal activity of the pest control. Caterpillar Publications, essential oils from three Nigerian medicinal Mariyappa Nagar, Tamil Nadu, India. plants, Conference Paper: 46th 1995;91. International Symposium on Essential Oils 38. Ajayi OE. Insecticidal activity of powder (ISEO) at Lublin, Poland. 2015;46. and extracts of Delonix regia seed against 29. Tahir HM, IshaqT, Mukhtar MK, Khan SY, maize weevil, Sitophilus zeamais Ahmed K. Potential use of Calotropis (Coleoptera: Curculionidae). Futa J. of procera (milk weed) to control Culex Res. Sci. 2013;1:54-62. quinquefasciatus (Diptera: Culicidae). 39. Sudhapriya A, Dhivya R. Phytochemical Pakistan J. Zool. 2013;45(3):615-621. screening and larvicidal efficacy of solvent 30. Butt A, Butt BZ, Vehra SE. Larvicidal extracts of Delonix regia leaf and flower potential of Calotropis procera against against vector mosquito Culex Aedes aegypti. Int. J. Mosq. Res. quinquefasciatus. Int. J. of Res. Rev. 2016;3(5):47-51. ISSN: 2348-5906. 2019;6(3):206-214. 31. Begum N, Sharma B, Pandey RS. DOI: inrein.com/10.4444/ijrr.1002/1029 Evaluation of insecticidal efficacy of 40. Olayemi IK, Samuel OM, Ukubuiwe AC, Calotropis procera and Annona squamosa Ande AT, Adeniyi KA, Shittu KO. Larvicidal ethanol extracts against Musca domestica. activities of leaf extracts of Adansonia J Biofertil. Biopestici. 2011;1:101. digitata L. (Malvales: Malvaceae) and DOI: 10.4172/2155-6202.1000101 Ficus sur Forssk (Rosales: Moraceae) 32. Traore A, Ouedraogo S, Belemlilga MB, against Culex quinquefasciatus mosquito Kabore A, Guissou IP. Phytochemical (Diptera: Culicidae). J. Mosq. Res. analysis and ovicidal activity of Cassia 2017;7(15):115-124. sieberiana, Guiera senegalensis and DOI: 10.5376/jmr.2017.07.0015 Excoecaria grahamii extracts. Afr. J. 41. Rahuman AA, Venkatesan P, Geetha K, Pharm. Pharmacol. 2017;11(44):554-560. Gopalakrishnan G, Bagavan A, Kamaraj C. Available:https://doi.org/10.5897/AJPP201 Mosquito larvicidal activity of gluanol 7.4837 acetate, a tetracyclic triterpenes derived 33. Kumar D, Chawla R, Dhamodaram P, from Ficus racemosa Linn. Parasitol. Res. Balakrishnan N. Larvicidal activity of 2008;103:333–339. Cassia occidentalis (Linn) against the DOI: 10.1007/s00436-008-0976-6 larvae of Bancroftian filariasis vector 42. Famuyiwa FG, Famuyiwa SO, Aladesanmi mosquito Culex quinquefasciatus. J. AJ. Activity of the compounds isolated Parasitol. Res. 2014;236838-236843. from Blighia sapida (Sapindaceae) Available:http://dx.doi.org/10.1155/2014/23 stem bark against Aedes aegypti 6838 larvae. Ife J. Sci. 2018;20(3):601- 34. Mbatchou VC, Tchouassi DP, Dickson RA, 605. Annan A, Mensah AY, Amponsah IK, Available:https://dx.doi.org/10.4314/ijs.v20i Jacob JW, Cheseto X, Habtemariam S, 3.13 Torto B. Mosquito larvicidal activity of 43. Mathumani M, Sunilson AJ. Preliminary Cassia tora seed extract and its key phytochemical and larvicidal activity of anthraquinones aurantio-obtusin and methanolic extract of selected plants obtusin. Parasit Vectors. 2017;10:562-570. against Aedes aegypti larvae. DOI: 10.1186/s13071-017-2512-y. Multidisciplinary Int. J. 2019;3:22-26. 35. Ullah Z, Ijaz A, Mughal TK, Zia K. 44. Nagappan P, Gomathinayagam S. Study Larvicidal activity of medicinal plant of mosquito larvicidal effects of Momordica extracts against Culex quinquefasciatus charantia (bitter gourd) extracts as Say. (Culicidae, Diptera). Int. J. Mosq. nanopowder. Int. J. ChemTech. Res. Res. 2018;5(2):47-51. 2014;6(9):4052-4054. ISSN: 0974-4290. 36. Aina SA, Banjo AD, Lawal OA, Okoh HI, 45. Kumar S, Wahab N, Mishra M, Warikoo R. Aina OO, Dedeke GA. The toxicity of Evaluation of 15 local plant species as extracts of Tetrapleura tetraptera (Aridan), larvicidal agents against an Indian strain of Delonix regia (Flame of the forest) and dengue fever mosquito, Aedes aegypti L. Raphia vinifera (Raffia palm) on the larvae (Diptera: Culicidae). Front. Physiol. of Anopheles gambiae. AJE. 2009;2(2):67- 2012;3(104):1-6. 70. ISSN: 1995-8994. DOI: 10.3389/fphys.2012.00104

Famuyiwa et al.; IJTDH, 41(11): 23-34, 2020; Article no.IJTDH.59818

46. Singh RK, Dhiman RC, Mittal PK. Mosquito Pharmaceutical Sci. 2014;2(12):1760- larvicidal properties of Momordica 1769. ISSN: 2321-3310. charantia Linn (Family: Cucurbitaceae). J. 57. Rawani A, Ghosh A, Chandra G. Mosquito Vect. Borne Dis. 2006;43:88–91. larvicidal activities of Solanum nigrum L. 47. Mekuria DB, Kashiwagi T, Tebayashi S, leaf extract against Culex quinquefasciatus Kim C. Cucurbitane glucosides from Say. Parasitol. Res. 2010;(5):1235-1240. Momordica charantia leaves as oviposition DOI: 10.1007/s00436-010-1993-9 deterrents to the leaf miner, Liriomyza 58. Rawani A, Ray AS, Ghosh A, Sakar M, trifolii. Biosci. Biotechnol. Biochem. Chandra G. Larvicidal activity of 2005;69(9):1706-1710. phytosteroid compounds from leaf extract DOI: 10.1515/znc-2006-1-21 of Solanum nigrum against Culex vishnui 48. Rwang PG, Effoim OE, Mercy KP, group and Anopheles subpictus. BMC Res Etokakan AM. Effects of Psidium guajava Notes. 2017;10:135-143. (guava) extracts on immature stage of DOI: 10.1186/s13104-017-2460-9 mosquito. IJCAM. 2016;4(5):132–137. 59. Patel MK, Tiwari A, Saxena VL. Larvicidal DOI: 10.15406/ijcam.2016.04.00132 activity of crude Solanum nigrum leaf and 49. Satyal P, Paudel P, Lamichhane B, Setzer berries extract against dengue vector- WN. Leaf essential oil composition and Aedes aegypti. Int. J. Curr. Res. Rev. bioactivity of Psidium guajava from 2018;10(14):16-21. Kathmandu, Nepal. Am. J. Essent. Oil. DOI:http://dx.doi.org/10.31782/IJCRR.2018 2016;3(2):11-14. ISSN: 2321-9114. .10144 50. Edwin UP, Nyiutaha IG, Essien AE, 60. Pereira TM, Silva VCB, Neto JAR, Alves Nnamdi OK, Sunday EM. Larvicidal effect SN, Lima LARS. Larvicidal activity of the of aqueous and ethanolic extracts of methanol extract and fractions of the green Senna alata on Anopheles gambiae, Culex fruits of Solanum lycocarpum (Solanaceae) quinquefasciatus and Aedes aegypti. Pak. against the vector Culex quinquefasciatus J. Pharm. Sci. 2013;26(3):561-566. (Diptera: Culicidae). Rev Soc Bras Med 51. Angaye TCN, Konmeze O, Seibokuro B, Trop. 2014;47(5):646-648. Adulphus P. Biolarvicidal control of malaria Available:http://dx.doi.org/10.1590/0037- using solvent extracts of Senna alata. MOJ 8682-0010-2014 Toxicol. 2019;5(1):40‒42. 61. Singha S, Chandra G. Mosquito larvicidal 52. Solomon PB, Oyedeji AA, Briyai FO, Bawo activity of some common spices and DDS, Abowei JFN. Biolarvicidal potentials vegetable waste on Culex of the methanolic-leaf-extracts of selected quinquefasciatus and Anopheles tropical plant species. J. Experimental Clin. stephensi. Asian Pac. J. Trop. Med. Toxicol. 2019;1(2):7-11. 2011;288-293. DOI:10.14302/issn.2641-7669.ject-19- DOI: 10.1016/S1995-7645(11)60088-6 2730 62. Sakthivadivel M, Gunasekaran P, 53. Abirami D, Murugan K. HPTLC Sivakumar M, Samraj A, Arivoli S, quantification of flavonoids, larvicidal and Tennyson S. Evaluation of Solanum smoke repellent activities of Cassia trilobatum L. (Solanaceae) aerial extracts occidentalis L. (Caesalpiniaceae) against for mosquito larvicidal activity against the malarial vector Anopheles stephensi Lis filarial vector Culex quinquefasciatus Say (Diptera: Culicidae). J. Phytol. 2011;3(2): (Diptera: Culicidae). J. Entomol. Zool. 60-72. ISSN: 2075-6240. Studies. 2014;2(6):102-106. ISSN: 2320- 54. Imam H. Evaluation of the larvicidal, 7078. antiplasmodial and cytotoxicity properties 63. Eze EA, Pierre S, Danga Y, Okoye FBC. of Cassia arereh Del. stem bark. Eur J. Larvicidal activity of the leaf extracts of Med Plants. 2013;3:78-87. Spondias mombin Linn. (Anacardiaceae) 55. Pavananundt P, Jiraungkoorskul K, Kosai from various solvents against malarial, P, Jiraungkoorskul W. Larvicidal properties dengue and filarial vector mosquitoes of Cassia siamea leaf against Aedes (Diptera: Culicidae). J. Vector Borne Dis. aegypti larvae. Int. J. Modern Agric. 2014;51:300-306. 2013;2(1):1-8. ISSN: 2305‐7246. 64. Ajaegbu EE, Danga SPY, Ikemefuna U, 56. Meenupriya J, Vinisha AS, Priya P. Cassia Chijoke IU, Okoye FBC. Mosquito alata and Cassia auriculata – Review of adulticidal activity of the leaf extracts of their bioactive potential. World J. of Spondias mombin L. against Aedes

Famuyiwa et al.; IJTDH, 41(11): 23-34, 2020; Article no.IJTDH.59818

aegypti L. and isolation of active principles. of early fourth instars of Helicoverpa J. Vector Borne Dis. 2016;53:17–22. armigera Hübner. Int. J. Insect Sci. 65. Ajaegbu EE, Younoussa L, Danga SPY, 2015;7:53–60. Chijoke IU, Okoye FBC. Mosquito repellent DOI: 10.4137/IJIS.S29127 activity of Spondias mombin L. 70. Upadhyay RK. Bio-efficacy of latex (Anarcadiaceae) crude methanol extract extracts from plant species Thevetia and fractions against Aedes aegypti L. J. nerifolia, and Artocarpus heterophyllus, Nat. Prod. Resour. 2016;7(3):240-244. Ficus glomerata and Calotropis procera on 66. Ray DP, Dutta D, Srivastava S, Kumar B, survival, feeding, development and Saha S. Insect growth regulatory activity of reproductive behaviour of Spodoptera Thevetia nerifolia Juss. against litura (F.) (Noctuidae: Lepidoptera). Int. J. Spodoptera litura (Fab.). J. Appl. Bot. Chem. Biochem. Sci. 2013;4:86-98. Food. Qual. 2012;85:212–215. 71. Nnamani CV, Oselebe HO, Ogbonna A. 67. Yadav S, Singh SP, Mittal PK. Toxicity of Effect of leaf extracts of Draceana aborea Thevetia peruviana (yellow oleander) L. and Vitex doniana sweet on the larvae against larvae of Anopheles stephensi and of Anopheles mosquito. Anim. Res. Int. Aedes aegypti vectors of malaria and 2008;5(2):835–837. dengue. J Entomol. Zool. Stud. DOI: 10.4314/ari.v5i2.48725 2013;1(6):85-87. ISSN: 2320-7078. 72. Nayak JB, Trajani B. Larvicidal activity of 68. Ramamurthy S, Umavathi S, Thangam Y, Vitex negundo leaf extract against Culex Revathi S, Sowmiya S, Thamaraiselvi A. quinquefasciatus mosquito larvae. Int. J. Lethal effect of Thevetia peruviana leaf Curr. Res. 2014;6(8):7983-7985. ISSN: extract on larval stages of Musca 0975-833X. domestica (L). Int. J. Adv. Res. Biol. 73. Chandrasekaran T, Thyagarajan A, Sci. 2015;2(11):165–170. ISSN: 2348- Santhakumari PG, Pillai AKB, Krishnan 8069. UM. Larvicidal activity of essential oil from 69. Mishra M, Kumar GKK, Kumar S. Impact of Vitex negundo and Vitex trifolia on dengue the stem extract of Thevetia neriifolia on vector mosquito Aedes aegypti. J. Braz. the feeding potential and histological Soc. Trop. Med. 2019;52:1-5. architecture of the midgut epithelial tissue DOI: 10.1590/0037-8682-0459-2018 ______© 2020 Famuyiwa et al.; This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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