Philippine Journal of Science 147 (3): 513-521, September 2018 ISSN 0031 - 7683 Date Received: 20 Feb 2018

Insecticidal Activity of Crude Ethanolic Extracts of Five Philippine Plants against Worm, pavonana Fabricius (: )

Abigaile Mia V. Javier1,*, Virginia R. Ocampo2, Flor A. Ceballo2, and Pio A. Javier2

1Agriculture Research Section, Atomic Research Division, Philippine Nuclear Research Institute- Department of Science and Technology, Commonwealth Avenue, Diliman, Quezon City 1101 Philippines 2Institute of Weed Science, Entomology and Plant Pathology, College of Agriculture and Food Science, University of the Philippines Los Baños, College, Laguna 4031 Philippines

Plant substances play a major role in pest management by exhibiting their insecticidal activity through toxicity, antifeedant activity, repellency, and growth regulatory activity. Ethanolic extracts from five plant species – “lantana”, Lantana camara (Linnaeus); oregano, Coleus amboinicus (Loureiro); “langkauas”, Alpinia pyramidata (Blume); “luyang dilaw”, Curcuma longa (Linnaeus); and “chichirica”, Catharanthus roseus (Linn.) – were evaluated for their insecticidal activities including contact and residual toxicity, antifeedant activity, and growth regulator activity against third larval instar of cabbage worm, Crocidolomia pavonana (Lepidoptera). Among the five ethanolic extracts, Cu. longa was the most toxic to cabbage worm when applied topically (LD50=51.00 µg/g) and through leaf residue film method (LC50=116.73 µg/mL) at 72 h after treatment. L. camara ranked second in providing contact toxicity and leaf residue film method. Ethanolic extract from Co. amboinicus showed high antifeedant activity against cabbage worm at 500 µg/mL. All the ethanolic extracts showed insect growth regulatory activities expressed in high larval and pupal mortalities for Co. amboinicus and Cu. longa; high number of abnormal adults for Ca. roseus; and shortened lifespan of adults in L. camara and A. pyramidata. In view of the overall pesticidal properties of the plants, ethanolic extracts from Cu. longa and L. camara can be exploited as botanical insecticides for cabbage worm management.

Key words: antifeedant, botanical insecticide, Curcuma longa, Lantana camara, topical toxicity

INTRODUCTION 2003). It is considered as one of the major constraints in production because the larvae feed on young Cabbage worm, Crocidolomia pavonana Fabricius leaves and attack the cabbage head that could lead to 100% (Lepidoptera: Crambidae), is one of the major pest of yield loss (Morallo-Rejesus & Navasero-Ward 2003). crucifers. Newly hatched larvae feed on the underside of the leaf and disperse throughout the other plant parts as The usual control employed against lepidopterous they mature. In some cases, they reach the center of the like cabbage worm is the use of synthetic pesticides. plant and produce mass webbing and frass (Smyth et al. However, continuous and improper use of chemical insecticides has several drawbacks; these include *Corresponding author: [email protected] insecticide resistance, toxicity to natural enemies, risk to

513 Philippine Journal of Science Javier et al.: Ethanolic Extracts Against Cabbage Worm Vol. 147 No. 3, September 2018 human health, and lower prices because of consumers' shift Plant Materials and Preparation of Ethanolic to buying organic products (Bommarco et al. 2011). The use Extracts of botanical pesticides is an effective alternative approach Fresh leaves of L. camara (Verbenaceae), Co. amboinicus for controlling many insect pests. Many plants have been (Lamiaceae), and Cymbopogon nardus Linn. (Poacaeae); reported to be toxic against cabbage worm. Methanolic rhizomes of A. pyramidata (Zingiberaceae) and Cu. longa extracts from seeds of Barringtonia asiatica, Melia (Zingiberaceae); and leaves, stems, and flowers of Ca. azedarach, and Annona muricata showed potential control roseus (Apocynaceae) were collected during Jun-Sep against C. pavonana. Mixture of these three plants at 0.1% 2012 in areas of Los Baños, Laguna. Plants collected caused 100% mortality at 3 days after application using were brought to the laboratory, washed with dechlorinated leaf disc method against second instar larvae (Retnasari et water, and air-dried under room temperature of 28±1 ºC. al. 2017). Mixture of ethyl acetate extract from leaves of The extraction procedure was conducted at the Insect Tephrosia vogelii, seeds of Brucea javanica and fruit from Physiology Laboratory of IWEP. The rhizomes were Piper aduncum (1:3:2.5) showed remarkable insecticidal chopped into small pieces, while the other plant parts activity against second instar larvae of C. pavonana with were detangled and cut into smaller pieces using scissors. LC50 value of 0.03% (Lina et al. 2013). In Indonesia, two The plant parts were oven-dried at 60 ºC for 48-72 h. The botanical insecticide formulations – mixtures (at 0.1%) air-dried plant parts were ground using the Nutri Tech of Piper retrofractum and An. squamosa, and mixtures of blender-juicer set at 32000 rpm. Two hundred grams (200 Aglaia ordorata and An. squamosa – were more effective g) of dried samples were kept in a clean bottle, soaked than the synthetic insecticide (deltamethrin) in reducing the in 500 mL of 95% ethanol for at least two weeks for population of C. pavonana in the field (Dadang et al. 2011). homogenization. After homogenization, the liquid mixture was filtered in Whatman filter paper No. 1 and the filtrate Although botanical extracts have been studied in many was concentrated using a rotary evaporator until all the laboratory tests, very few studies are available on study solvent had evaporated. The resulting ethanolic extracts of insecticidal activities of ethanolic extracts against were stored at 4 °C in airtight bottles until use in bioassay. cabbage worm. Therefore, in this study, the researchers reported the insecticidal activity of ethanolic extracts of five Philippine medicinal plants including Alpinia pyramidata, Preparation of Test Solutions Lantana camara, Coleus amboinicus, Curcuma longa, and Stock solutions and lower concentrations were prepared Catharanthus roseus against cabbage worm. Most of them for each ethanolic extracts using acetone as diluent. The were reported toxic against stored pests and mosquitoes. final doses and concentrations for each plant extract were These plants also showed promising insecticidal activity prepared from the stock solution using acetone as diluent. using aqueous, alcoholic extracts using coconut wine Final doses and concentrations against cabbage worm (Javier et al. 2015), and essential oil extracts against Plutella were determined from preliminary bioassays. Series of xylostella Linnaeus (Javier et al. 2016) and Spodoptera dilutions between doses and concentrations that caused litura Fabr. (Javier et al. 2017). To evaluate the insecticidal 10-90% mortality of test insect was identified from the activity of ethanolic extracts of five Philippine medicinal preliminary results. plants (A. pyramidata, L. camara, Co. amboinicus, Cu. longa, and Ca. roseus), the researchers determined the Bioassay Procedures contact toxicity, residual toxicity, antifeedant property, and Contact Toxicity Test: Topical Application Method. morphogenetic effects against third instar larvae of cabbage One microliter (1 µL) of the solution of each treatment worm. The study was conducted at LB Uichanco Wing, was topically applied equivalent to doses of 8, 15, 31, Institute of Weed Science, Entomology and Plant Pathology 61, 122, and 245 µg/g larva on the thoracic region of (IWEP), College of Agriculture and Food Sciences (CAFS), each larva using a Hamilton Repeating Dispenser plus University of the Philippines, Los Baños (UPLB), College, a 10-µL microsyringe. Acetone and chlorfluazuron Laguna from Jun 2012 to Nov 2013. (Atabron 5 EC) at a dose of 12 µg/g larval body weight (recommended rate) served as the negative and positive controls, respectively. Ten larvae were placed in a Petri dish and were given 8 cm-diameter of pechay leaf as food. MATERIALS AND METHODS Ten larvae were used per replication and each treatment was replicated four times. In all the tests, third instar larvae Rearing of C. pavonana of cabbage worm with an average weight of 4.089 mg per Larvae of cabbage worm were collected from pechay larva were used. Mortality was recorded at 24, 48, and 72 plants, (Loureiro), grown around the h after treatment (HAT) (Javier et al. 2016; Javier et al. greenhouse of the IWEP, UP Los Baños and were reared in 2017; Javier et al. 2018). the laboratory following the method of Javier et al. (2018).

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Residual Toxicity Test: Leaf Residue Film Method. An the LD25 of the ethanolic extracts based on contact toxicity 8 cm-diameter pechay leaf was cut and 0.50 mL of the test of the five extracts using topical application at 72 HAT. different test solutions (31.25, 62.5, 125, 250, 500, and Third instar cabbage worm larvae were topically treated 1000 µg/mL) was evenly spread on both abaxial and with 30.57 μg/g body weight of the ethanolic extracts. adaxial sides of the leaves using forceps. After air-drying for about 10 min, each treated leaf was placed in a Petri Each of the treated larvae was released inside a Petri dish dish measuring 9 cm, and ten third instar larvae of cabbage containing moistened filter paper and pechay leaves as food. worm were introduced (Javier et al. 2016; Javier et al. Each treatment was replicated four times. Pechay leaves were 2017; Javier et al. 2018). Each treatment was replicated provided to the larvae until pupation and left-over leaves and four times. Acetone and chlorfluazuron at a concentration excreta were removed daily. The mortality and abnormalities of 50 µg/mL (recommended rate for cabbage worm) were observed until adult emergence (Pipithsangchan & served as the negative and positive controls, respectively. Morallo-Rejesus 2005; Javier et al. 2016). Mortality was recorded at 24, 48, and 72 HAT. Statistical Analysis Antifeedant Activity Test (No-choice Test). The pechay For contact and residual toxicity tests, mortality recorded leaf was treated as described in the residual toxicity test. at 24, 48, and 72 HAT were subjected to probit analysis After drying, each leaf disc was placed in the Petri dish. Ten using PoloPlus© developed by LeOra Software in 2002 third instar larvae of cabbage worm previously starved for (Finney 1971) to determine the LD or LC (dose or 6 h were introduced on to the leaf disc. Each treatment was 50 50 concentration that gives 50% mortality on the test larvae). replicated four times. Acetone and ethanolic extract from For the relationship between dose and response data on Cy. nardus with concentrations same with plant extracts mortality and antifeedant activity, Pearson’s coefficients (31.25, 62.5, 125, 250, 500, and 1000 µg/mL) served as were calculated and tested. The response data were the control and standard repellent, respectively. Extracts subjected to one-way analysis of variance (ANOVA). from Cy. nardus was used for over fifty years both as Means were separated using Tukey’s Honest Significant antifeedant and insect repellent against many insect pests. Difference (HSD) for further analysis to know the The biocidal activity of ethanolic extract from Cy. nardus significant difference among the treatments. Differences was mainly attributed to terpene hydrocarbons such as were considered significant at P<0.05. monoterpenes and sesquiterpenes, and oxygen derivatives such as aldehydes and alcohols (Doumbia et al. 2014). The amount of feeding in each treatment was based on percent reduction in weight of the pechay leaf consumed RESULTS AND DISCUSSION by the larvae after 24 h. The amount of feeding was recorded by weighing the leaf before and after the Contact Toxicity: Topical Application Method consumption. The corrected antifeedant index (Blaney During the entire observation, Cu. longa was the most et al. 1990) was calculated as follows: toxic against cabbage worm. At 72 HAT, Cu. longa (LD50 C-T value of 51.00 µg/g) and L. camara (LD50 value of 53.99 % Relative Feeding Index or RFI = x 100 (1) C+T µg/g) were significantly more toxic than Co. amboinicus, A. pyramidata and Ca. roseus against cabbage worm and where C is leaf consumption in control, and T is leaf ranged from 51.00 to 108.82 µg/g against cabbage worm consumption in treated. The RFI measures the feeding using topical method (Table 1). deterrence of each test solution. Results of bioassay of different doses of the ethanolic Antifeedant activity was evaluated using the following extracts are graphically represented in Fig. 1. Based on scale (Huang et al. 2000): the dose-survival curves of all the plant extracts, the. L. camara was the most toxic followed by Cu. longa. The % RFI AntiFeedant activity ethanolic extracts from Co. amboinicus and Cu. longa > 75 High showed almost the same toxicity against cabbage worm. 51 – 74 Moderate Comparison of the activity of the ethanolic extracts with 25 – 50 Low chlorfluazuron, suggests that L. camara and Cu. longa < 24 Very weak at 122 and 245 µg/g body weight provided comparable mortality with chlorfluazuron at 72 HAT (Table 2). The Test on Morphogenetic Effects. The insect growth other ethanolic plant extracts were less toxic than the regulator (IGR) activity of the plant extracts was determined chlorfluazuron, because it is not pure synthetic derived using ten third instar larvae of cabbage worm, which were chemical unlike the commercial insecticides. pre-weighed prior to exposure. The larvae were treated with

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Table 1. Lethal dose (LD50) (µg/g larval body weight) of five ethanolic extracts against third larval instar of Crocidolomia pavonana at 24, 48, and 72 h after topical treatment1. Plant species 24 HAT 48 HAT 72 HAT Coefficient A. pyramidata 1184.30 (385–38815)c 300.67 (177–878)c 108.63 (76–175)b y=0.0672x+11.82, R2=0.9448 L. camara 142.91 (100–238)b 80.98 (61–113)a 53.99 (35–85)a y=0.0822x+17.24, R2=0.8190 Co. amboinicus 359.90 (143–10982)b 187.98 (116–452)bc 107.99 (59–357)b y=0.0597x+16.05, R2=0.8852 Cu. longa 264.60 (137–1127)b 140.92 (86–332)b 51.00 (36–73)a y=0.0715x+21.68, R2=0.8345 Ca. roseus 250.91 (151–634)b 242.57 (137–783)bc 108.82 (66–253)b y=0.0661x+12.49, R2=0.9388

1 LD50= dose that results in 50% mortality of the test larvae; CI= class index at 95% confidence interval; HF = heterogeneity factor. For each species, values followed by the same letter in the same column are not significantly different because of the overlapping of the confidence intervals (P<0.05).

Table 2. Percent mortality of third instar larvae of Crocidolomia pavonana topically applied with different doses of ethanolic extracts at 72 h after treatment1. Dose (µg/g) A. pyramidata L. camara Co. amboinicus Cu. longa Ca. roseus T1 – 8 15.0 ef 12.5 cd 15.0 ef 20.0 c 15.0 de T2 – 15 12.5 ef 25.0 c 27.5 de 32.5 c 15.0 de T3 – 31 27.5 de 27.5 c 32.5 d 32.5 c 30.0 d T4 – 61 32.5 d 47.5 b 30.0 de 55.0 b 30.0 d T5 – 122 50.0 c 82.5 a 50.0 c 65.0 b 50.0 c T6 – 245 75.0 b 85.0 a 72.5 b 85.0 a 77.5 b T7 – Control 2.50 f 2.50 d 2.50 f 2.50 d 2.50 e T8 – Chlorfluazuron (12) 97.50 a 1Based on 10 larvae per replicate; each dose was replicated four times. Means followed by the same letter(s) within the same column are not significantly different at 5% level of Tukey’s HSD test.

Figure 1. Survival of third instar larvae of Crocidolomia pavonana topically applied with different doses of ethanolic extracts at 72 h after treatment.

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The insecticidal constituent of Cu. longa rhizome was film method. At 72 HAT, Cu. longa and L. camara showed identified as sesquiterpene ketone ar-turmerone by LC50 values of 116.73 and 118.94 µg/mL, respectively spectroscopic analysis, which was found effective against (Table 3). P. xylostella and Nilaparvata lugens Stal (Lee et al. 2001). Results of bioassay of different concentrations are Essential oil from Cu. longa showed LD50 values of 32.98 µg/g, 792 µg/g, and 308.05 µg/g against P. xylostella, S. graphically represented in Fig. 2. Based on the litura and C. pavonana, respectively at 72 HAT (Javier et concentration-survival curves of all the plant extracts, the. al. 2016; Javier et al. 2017; Javier et al. 2018). Meanwhile, L. camara and Cu. longa were the most toxic, followed by Javier et al. (2018) reported that essential oil from L. A. pyramidata. The activity of the different concentrations camara exhibits insecticidal against cabbage worm with of the ethanolic extracts from five plant species was compared with the conventional insecticide benzoylurea LD50 value of 169.83 µg/g. Spraying of ethanolic extract from leaves of L. camara increased yield of cabbage plants recommended against cabbage worm (chlorfluazuron, by 25.80% (Baidoo & Adam 2012). Atabron 5 EC). The three lowest concentrations of the ethanolic extracts (31.25, 62.5, and 125 µg/mL) were quite ineffective providing less than 50% cabbage worm Residual Toxicity Test: Leaf Residue Film Method larval mortality (Table 4). The application of 500 µg/mL Based on the LC50 values, Cu. longa and L. camara of L. camara, Cu. longa, and A. pyramidata provided at showed the highest toxicity against cabbage worm at all least 90% mortality at 72 HAT, which did not significantly hours of observation when applied through leaf residue differ from 50 µg/mL of chlorfluazuron. Co. amboinicus

1 Table 3. Lethal concentration (LC50) (µg/mL) of five ethanolic extracts against Crocidolomia pavonana at 24, 48, and 72 h after treatment . Plant species 24 HAT 48 HAT 72 HAT Coefficient A. pyramidata 565.98 (323–1721)ab 299.15 (209–469)c 141.25 (81–230)ab y= 0.0888x+23.23, R2=0.7950 L. camara 257.28 (159–461)a 129.50 (83–195)a 118.94 (72–183)a y= 0.0938x+25.40, R2=0.7401 Co. amboinicus 389.28 (219–1063)a 216.06 (168–282)b 187.47 (145–241)b y= 0.0852x+19.25, R2=0.7988 Cu. longa 414.05 (213–1127)ab 150.34 (76–281)b 116.73 (74–173)a y= 0.0941x+25.67, R2=0.7143 Ca. roseus 745.88 (434-2306)b 448.96 (339–647)d 272.28 (222–334)c y= 0.0965x+8.56, R2=0.8930

1 LC50= concentration which results in 50% mortality of the test larvae; CI= class index at 95% confidence interval; HF = heterogeneity factor. For each species, values followed by the same letter in the same column are not significantly different because of the overlapping of the confidence intervals (P<0.05).

Figure 2. Survival of third instar larvae of Crocidolomia pavonana treated by leaf residue film method with different concentrations of ethanolic extracts at 72 h after treatment.

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and Ca. roseus showed mortality comparable with Alpinia galangal showed toxicity against adult B. dorsalis chlorfluazuron when concentration was increased to Hendel with LC50 values of 5987.05 and 5652.53 ppm at 1000 µg/mL. 24 and 48 h, respectively (Sukhirun et al. 2009). Essential oil extracts from rhizomes of A. pyramidata showed Field application of petroleum ether extracts of L. camara LC50 values of 283.86 µg/mL and 33.3 mg/mL against P. leaves in cabbage significantly reduced the number of xylostella and S. litura, respectively at 72 HAT (Javier et pests including P. xylostella, aphids Brevicoryne brassicae al. 2016; Javier et al. 2017). Linn., and webworm Hellula undalis Fabr. on cabbage, resulting to 25.80% increase yield when applied at 10 mL/L of water (Baidoo & Adam 2012). Female adults Antifeedant Activity of peach fruit fly, Bactrocera zonata Saunders showed Among the ethanolic extracts, Co. amboinicus was 85, 67, and 57% mortality when fed for 16 days on a diet comparable with the standard antifeedant Cy. nardus which containing acetone extract from rhizomes of Cu. longa at showed high RFI at 500 µg/mL, followed by Cu. longa 1000, 500, and 250 ppm, respectively (Siddiqi et al. 2011). which exhibited high RFI at the highest concentration of 1000 µg/mL. L. camara exhibited moderate RFI at 250 µg/mL, Meanwhile, there are very limited researches on the while A. pyramidata exhibited moderate RFI at 500 µg/mL. insecticidal activity of the ethanolic extracts from A. All concentrations of the ethanolic extract from Ca. roseus pyramidata. Methanolic extracts from the rhizomes of consistently showed low RFI against cabbageworm (Table 5).

Table 4. Percent mortality of different concentrations (µg/mL) of ethanolic extracts against third instar larvae of Crocidolomia pavonana at 72 h after treatment by leaf residue film method1. Concentration ( µg/mL) A. pyramidata L. camara Co. amboinicus Cu. longa Ca. roseus T1 - 31.25 12.5 d 12.50 d 12.50 d 12.50 d 2.50 e T2 - 62.5 35.0 c 30.00 c 17.50 d 35.00 c 7.50 e T3 – 125 47.5 bc 52.50 b 45.00 c 37.50 c 25.00 d T4 – 250 52.5 b 65.00 b 52.50 c 80.00 b 42.50 c T5 – 500 90.00 a 100 a 82.50 b 97.50 a 77.50 b T6 – 1000 97.50 a 100 a 90.00 ab 100 a 90.00 ab T7 - Control 2.50 d 2.50 d 2.50 d 2.50 d 2.50 e T8 - Chlorfluazuron 100 a (50 µg/mL) 1Based on 10 larvae per replicate; each dose was replicated four times. Means followed by the same letter(s) within the same column are not significantly different at 5% level of Tukey’s HSD test.

Table 5. Percent relative feeding index (RFI)1 of the second larval instar of Crocidolomia pavonana at 24-h exposure to the cabbage leaf treated with different concentrations of ethanolic extracts. Concentrations (µg/mL)2 Plant Species 125 250 500 1000 Coefficient A. pyramidata 38.92 (L) 49.43 (L) 61.17 (M) 60.38 (M) y=0.0218x+42.25, R2=0.6455 L. camara 38.10 (L) 61.51 (M) 62.74 (M) 63.33 (M) y=0.0202X+46.97 R2=0.4062 Co. amboinicus 52.93 (M) 61.71 (M) 79.53 (H) 81.25 (H) y=0.0311x+54.29, R2=0.7576 Cu. longa 35.29 (L) 62.43 (M) 72.57 (M) 80.03 (H) y=0.0428x+44.40, R2=0.6133 Ca. roseus 28.72 (L) 29.72 (L) 32.40 (L) 40.26 (L) y=0.0134x+26.49, R2=0.9869 Cy. nardus 69.97 (M) 62.43 (M) 78.77 (H) 86.56 (H) y=0.0239x+63.23, R2=0.7781 1RFI = [(C-T) / (C+T)] x 100, where: C= leaf consumption in control; T= leaf consumption in treated. The RFI measures the feeding deterrence of each test solutions. % RFI Antifeedant activity > 75 High (H) 51 – 74 Moderate (M) 25 – 50 Low (L) < 24 None (N) 2Based on 10 larvae per replicate; each concentration was replicated four times.

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Essential oil from leaves of Co. amboinicus showed high pyramidata. Meanwhile, all the other ethanolic extracts antifeedant activity with RFI of 87.68% against second did not significantly differ with the control (Table 6). instar P. xylostella at 500 µg/mL (Javier et al. 2016). Meanwhile, essential oil from leaves of Co. amboinicus Results suggest that ethanolic extracts of the plants showed moderate RFI of 53.47% against third instar larvae exhibited IGR activity against the third instar larvae of of cabbageworm at 4000 µg/mL. cabbage worm, especially the ethanolic extract from Co. amboinicus and Cu. longa which showed high larval Dichloromethane extract from rhizomes of Cu. longa mortality, larval-pupal intermediates, and abnormal pupae; showed high antifeedant activity against second and third Ca. roseus which showed high number of abnormal adults; instar larvae of Helicoverpa armigera Hubner, but did and L. camara and A. pyramidata which shortened the not exhibit ovicidal and larvicidal activity (Hernandez lifespan of adults treated in larval stage. 2003). Essential oil from rhizomes of Cu. longa showed 100% antifeedant activity against second instar larvae of In a previous study, essential oils from leaves of Co. P. xylostella at 500 µg/mL (Javier et al. 2016). Meanwhile, amboinicus also showed morphogenetic effects against oil from rhizomes of A. pyramidata showed high RFI second instar larvae of P. xylostella as expressed in 30% against S. litura when concentration is at 8 mg/mL (Javier larval mortality, 16.67% abnormal pupae among the alive et al. 2017). larvae, and 35% abnormal adults among the alive pupae (Javier et al. 2016). Topical application of essential oils from Cu. longa showed high number of abnormal pupae Insect Growth Regulatory Activity: Morphogenetic and adults against S. litura (Javier et al. 2017). The active Effects ingredient, 2-phellandrene from essential oil of Cu. longa The cabbage worm was treated topically with 30.57 μg/g acts as IGR on hairy caterpillar, Spilosoma obliqua body weight of ethanolic extracts. Among the ethanolic Walker (Agarwal et al. 1999). Meanwhile, essential oil extracts, highest larval mortality of 42.50% was obtained from leaves of L. camara provided high larval 60% larval from Co. amboinicus and Cu. longa. These extracts mortality and 12.50% pupal mortality from larvae that also provided 12.5% larval-pupal intermediates, which survived, when third instar larvae of S. litura was treated is the highest among the treatments. From the larvae (Javier et al. 2017). that survived, there were about 16.67 to 38.89% dead or abnormal pupae; the highest of which was observed from the larvae treated with Cu. longa followed by Co. amboinicus (33.33%). From the normal pupae, 10- CONCLUSION 41.18% emerged as abnormal adults; highest of which was observed from the larvae treated with Ca. roseus – Among the five ethanolic extracts, Cu. longa was the most followed by Cu. longa (36.36%). The normal pupae of toxic against cabbage worm (LD50 = 51.00 µg/g) through the cabbage worm emerged as adult and lived for topical application, which did not significantly differ from 7-14 days among all the treatments including the control, L. camara (LD50 = 53.99 µg/g) at 72 HAT. When applied while the abnormal pupae did not emerge at all. Among through LRFM, Cu. longa was still the most toxic against the ethanolic extracts, shortest adult lifespan was recorded cabbage worm (LC50 = 116.73 µg/mL), which did not on cabbage worm treated with L. camara, which did significantly differ from L. camara (LC50 = 118.94 µg/m). not significantly differ with the lifespan recorded in A. Co. amboinicus showed high antifeedant activity against cabbage worm at 500 µg/mL. Cu. longa also showed high

Table 6. Effects of ethanolic extracts on the development of cabbage worm at 31 μg/g1,2. Plant extracts L-mortality3 L-P mortality3 Survival Abnormal Pupae Abnormal Adults Longevity (31 μg/g) (%) (%) (%) (%) (%) (days) A. pyramidata 32.50 7.50 60.00 16.67 10.00 10.22 b L. camara 35.00 2.50 62.50 24.00 10.53 9.82 b Co. amboinicus 42.50 12.50 45.00 33.33 25.00 12.00 a Cu. longa 42.50 12.50 45.00 38.89 36.36 11.71 a Ca. roseus 40.00 5.00 55.00 22.73 41.18 11.70 a Control 2.50 2.50 95.00 2.63 5.41 12.23 a 1Based on 10 larvae per replicate; each concentration was replicated four times 2Means followed by the same letter(s) within the same column are not significantly different at 5% level of Tukey’s HSD. 3L- larval; L-P- larval-pupal

519 Philippine Journal of Science Javier et al.: Ethanolic Extracts Against Cabbage Worm Vol. 147 No. 3, September 2018 antifeedant activity when concentration was increased DOUMBIA M, YOBOUE Y, KOUAMÉ LK, COFFI K, to 1000 µg/mL. Co. amboinicus and Cu. longa showed KRA DK, KWADJO KE, DOUAN BG, DAGNOGO major IGR activities against cabbage worm as expressed M. 2014. Toxicity of Cymbopogon nardus (Glumales: in high larval and pupal mortalities. Ca. roseus showed Poacea) against four stored food products insect pests. high number of abnormal adults, while L. camara and Intl J Farm & Alli Sci 3(8): 903–909. A. pyramidata shortened the lifespan of adults treated in FINNEY DT. 1971. Probit analysis, 3rd Ed. Cambridge larval stage. In view of the overall pesticidal properties University Press, London. 383p. of the plants, ethanolic extracts from Cu. longa and L. camara can be exploited as botanical insecticides for HERNANDEZ HP. 2003. Insecticidal components from cabbage worm management. luyang dilaw, Curcuma longa L.: Antifeedants to Crocidolomia binotalis Zeller [2003]. Univ. Los Banos, College, Laguna (Philippines). Inst. of Chemistry. HUANG Y, LAM SL, HO SH. 2000. Bioactivities of ACKNOWLEDGMENT essential oil from Elletaria cardamomum (L.) Marton We thank the following: Dr. Aurora M. Baltazar, to Sitophilus zeamais Motschulsky and Tribolium guidance committee member for her comments and castaneum (Herbst.). J Stored Prod Res 36: 107–117. invaluable suggestions in improving this work; DOST JAVIER PA, PADILLA CL, PUNZALAN EG. 2015. – Science Education Institute – Accelerated Science and Development of insect pest management products and Technology Human Resource Development Program for systems for organic vegetable production in Southern providing the research funds to pursue this study; and Luzon. DOST-PCAARRD Terminal Report, 135p. staff of Department of Entomology of Institute of Weed Science, Entomology and Plant Pathology, UPLB for their JAVIER AMV, OCAMPO VR, CEBALLO FA, JAVIER assistance during this research. PA. 2016. Insecticidal activity of four essential oils against diamondback moth, Plutella xylostella Linnaeus (Lepidoptera: Pyralidae). Philipp Agric Sci 99(2): 156–163. REFERENCES CITED JAVIER AMV, OCAMPO VR, CEBALLO FA, JAVIER AGARWAL M, WALIA S, DHINGRA S. 1999. Pest PA. 2017. Insecticidal activity of selected essential control properties of turmeric leaf oil against Spilosoma oils extracts against common cutworm, Spodoptera obliqua, Dysdercus koenigii and Tribolium castaneum. litura Fabricius (Lepidoptera: Noctuidae). Philipp J Proceed. 2nd All India People’s Congress, Calcutta. Sci 146(3): 247–256. p. 1–7. JAVIER AMV, OCAMPO VR, CEBALLO FA, JAVIER BAIDOO PK, JI ADAM. 2012. The effects of extracts PA. 2018. Insecticidal activities of the essential of Lantana camara (L.) and Azadirachta indica oils from different plants against cabbage worm, (A. Juss) on the population dynamics of Plutella Crocidolomia pavonana Fabricius (Lepidoptera: xylostella, Brevicoryne brassicae and Hellula undalis, Crambidae). Philipp Agric Sci. In Press. on cabbage. Sust Agri Res 1(2): 229–234. LEE HS, SHIN WK, SONG C, CHO KY, AHN YJ. 2001. BLANEY WM, SIMMONDS SJ, LEY SV, ANDERSON Insecticidal activities of ar-turmerone Identified in S, TOOGOOD PL. 1990. Antifeedant effects of Curcuma longa rhizome against Nilaparvata lugens azadirachtin and structurally related compounds on (Homoptera: Delphacidae) and Plutella xylostella lepidopterous larvae. Entomologia Experimentalis et (Lepidoptera: Yponomeutidae. J Asia Pac Entomol Applicata 55: 149–160. 4: 181–185. BOMMARCO R, MIRANDA F, BYLUND H, LINA EC, DADANG, MANUWOTO S, SYAHBIRIN BJORKMAN C. 2011. Insecticides suppress natural G, PRIJONO D. 2013. Synergistic action of mixed enemies and increase pest damage in cabbage. J Econ extracts of Brucea javanica (Simaroubaceae), Entomol 104(3): 782–791. Piper aduncum (Piperaceae), and Tephrosia vogelii (Leguminosae) against cabbage head caterpillar, DADANG, FITRIASARI ED, PRIJONO D. 2011. Field Crocidolomia pavonana. J Biopest 6(1): 77–83. efficacy of two botanical insecticide formulations against cabbage insect pests, Crocidolomia pavonana MORALLO-REJESUS B, NAVASERO-WARD R. 2003: (F.) (Lepidoptera: Pyralidae) and Plutella xylostella Biology of Crocidolomia pavonana (F.) (Lepidoptera: (L.) (Lepidoptera: Yponomeutidae). J ISSAAS 17(2): Pyralidae) and population interventions against this 38–47. recurrent pest of crucifers in the Philippines. Philipp

520 Philippine Journal of Science Javier et al.: Ethanolic Extracts Against Cabbage Worm Vol. 147 No. 3, September 2018

Ent 17(2): 141–168. PIPITHSANGCHAN S, MORALLO-REJESUS B. 2005. Insecticidal activity of diosgenin isolated from three species of grape ginger (Costus spp.) on the diamondback moth, Plutella xylostella (L.) Philipp Agric Sci 88(3): 317–327. RETNASARI E, PUSPASARI LT, MELIANSYAH R, MAHARANI R, HIDAYAT Y, DONO D. 2017. Toxicity of Barringtonia asiatica L. (Kurz.), Melia azedarach L. and Annona muricata L. seed extract mixture against larvae Crocidolomia pavonana F. (Lepidoptera:Pyralidae). 2nd International Conference on Sustainable Agriculture and Food Security: A Comprehensive Approach. KnE Life Sciences Nov 2017: 246–255. SIDDIQI AR, NAZ F, MASIH R, AHMAD I, JILANI G. 2011. Effects of Curcuma longa extracts on mortality and fecundity of Bactrocera zonata (Diptera: Tephritidae). Ciência e Agrotecnologia. 35(6): 1110–14. SMYTH RR, HOFFMANN MP, SHELTON AM. 2003. Effects of host plant phenology on oviposition preference of Crocidolomia pavonana (Lepidoptera: Pyralidae). Environ Entomol 32(4): 756–764. SUKHIRUN N, BULLANGPOT V, PLUEMPANUPAT W. 2009. The insecticidal studies from Alpinia galanga and viscosa extract as alternative control tool to Bactrocera dorsalis (Hendel). KKU Sci J (Supplement) 37: 71–76.

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