Vol. 7(36), pp. 2645-2656, 25 September, 2013

DOI: 10.5897/JMPR11.1662

ISSN 1996-0875 ©2013 Academic Journals

http://www.academicjournals.org/JMPR

Journal of Medicinal Plants Research

Full Length Research Paper

Comparative toxicity of Azadirachta indica A. juss. and Callistemon citrinus D. C. against sugarcane stalk borer

Chilo auricilius Dudgeon (Lepidoptera: Crambidae)

Rashid Mumtaz1*, Anwar L. Bilgrami2 and Saleh A. Aldosari1

1Plant Protection Department, College of Food and Agricultural Sciences, King Saud University, P.O.Box.2460 Riyadh-

11451, Saudi Arabia. 2Cape May County Department of Mosquito Control, P. O. Box. 66, Cape May Court House, New Jersey 08210, USA.

Accepted 14 February, 2012

Efficacy of Azadirachta indica A. Juss. leaves and Callistemon citrinus D.C. leaves extract were tested against sugarcane stalk borer, Chilo auricilius Dudgeon. The plant extract prepared by sequential extraction method using different solvent like, petroleum, chloroform, methanol, distilled water and acetone at different per cent concentrations were used. Ovipositional behavior of adult moth and hatching percentage of the eggs were recorded using choice method for each fraction of the plant extract. The methanol extracts of A. indica 12.33% eggs/female and chloroform extracts of C. citrinus 13.33% eggs/female were found to be effective, and reduced oviposition as compared to other extracts. At 20% concentration, methanol extracts of A. indica and chloroform extract of C. citrinus gave lowest larval survival (55, 52.33 and 48.66% larval survival) after 24, 48 and 72 h of exposures, respectively. However, petroleum ether extract of A. indica at 8, 10 and 20% concentration survival of three days old eggs were registered at 60, 55 and 52%; whereas chloroform extract of C. citrinus at similar concentration recorded 70, 64 and 61% survival. Key words: Plant extracts, neem, bottle brush, sugarcane stalk borer.

INTRODUCTION Economic interest in sugarcane Saccharum officinarum L. has increased significantly in recent years due to the increased worldwide demand for sustainable energy production. Sugarcane is a monocotyledonous crop, and cultivated in tropical and subtropical regions of the world. Sugarcane crop incur losses by insect pests under modern agriculture (Dhaliwal and Arora, 1996; Dhaliwal et al., 2007). Though, insects are a major biotic constraint in sugarcane crop production particularly in subtropics that provides a conducive environment for insect pest built up, the major pest species of subtropics are Chilo infuscatellus Snellen, Scirpophaga excerptatis Walker and Chilo auricilius Dudgeon.

C. auricilius causes damage early during growth, killing leaves and possibly producing 'dead hearts'. Sugarcane trials of Trichogramma spp. with the exotic tachinid Lixophaga diatraeae had failed to give effective control of C. auricilius (Chaudhary and Sharma, 1981). Therefore, the evaluation of the bio-efficacy of plant extracts is difficult in respect of the target pest. Pest control strategies from unilateral chemical approach to non-chemical methods such as bio pesticides, biological control, ecological management and host plant resistance has, therefore, become imperative. While, the indiscri-minate use of chemical pesticides has given too many serious problems, including genetic resistance of pest

*Corresponding author. E-mail: rashidmumtazkhan@gmail.com.

2646 J. Med. Plants Res. species, chemical pesticides have brought in imbalances in biotic system resulting into development of loss of bio-diversity and resurgence of pests and secondary pest outbreaks apart from environmental pollution, contami-nation of food chain and human and animal health hazards (Ahmed et al., 1981; Arthur, 1994; Swarnasiri and Palipane, 1995). In the light of aforementioned, there is a world-wide interest in the development of alternative strategies and search for new types of insecticides (Bernays, 1983; Berenbaum, 1985; Tripathi et al., 2000). Botanical pesticides are highly effective, safe and ecologically acceptable (Ryan, 2002; Senthil Nathan and Kalaivani, 2005; Isman and Akhtar, 2007). Use of A. indica based on botanical pesticides has been reported for protecting agricultural crops (Ascher and Meisner, 1989; Schmutterer, 1990; Mordue and Blackwell, 1993; Pearsall and Hogue, 2000; Vinuela et al., 2000; Mordue, 2004). More than 140 active principals have been identified in the A. indica tree (Koul, 2004). Callistemon species are also used for forestry, essential oil production, farm tree/windbreak plantings, degraded-land reclamation, and ornamental horticulture among other applications (Spencer and Lumley, 1991). In China, Callistemon species, especially C. viminalis, are used in traditional Chinese Medicine pills for the treatment of hemorrhoids (Ji, 2009). Callistemon species, used as wild plant control and as bio-indicators for environmental management (Burchett and Tarran, 2002; Wheeler, 2005). Earlier phytochemical explorations of members of this genus resulted in the identification of C-methyl flavonoids, triterpenoids and phloroglucinol derivatives (Huq and Misra, 1971; Varma et al., 1975; Younes, 1975; Wrigley and Fagg, 1993; Wollenweber et al., 2000). Besides, some medicinal properties like antimicrobial, anti-staphylococcal, antithrombin, repellent and nematici-dal activities as well as larvicidal and pupicidal values have been reported for the genus (Sangwan et al., 1990; Chistokhodova et al., 2002; Aisien et al., 2004; Kobayashi et al.,2006; Saxena and Gomber, 2006; Gomber, 2007). In South Africa, Callistemon species are grown as garden, street trees or ornamental plants because of their decorative flowers. C. citrinus is the most widely cultivated member of the genus Callistemon. The effects of some botanical pesticides on insect pests, such as repellent, antifeedent and toxic effects are shown, but few are not favorable (Karelina et al., 1992; Naumann and Isman, 1995; Prakash and Rao, 1997; Päts and Isman, 1998; Govindachari et al., 2000; Meadow and Seljasen, 2000; Turcani, 2001; Koul, 2004; Regnault-

Roger et al., 2005; Isman, 2006). These controversial results suggest that the efficacy of any A. indica -based insecticides on pests should be investigated before widespread use in the field.

Application of A. indica and C. citrinus plant extracts against C. auricilius was investigated in the laboratory. The plants included in the study are abundantly available and easily accessible in India. Using plant extracts for management of C. auricilius will on one hand reduce

dependence on synthetic insecticides, and on the other hand help in maintaining a desirable ecological balance. C. auricilius selected as the test insect mainly because of its wide host range. MATERIALS AND METHODS

Bio efficacy of natural plant extracts against the sugarcane stalk borer was investigated under the laboratory conditions. Adult C.

auricilius were collected from field sugarcane of North State of India. These adults were reared in the laboratory with help of standard rearing technique. Ten pairs of freshly emerged moths were released for egg-laying on potted sugarcane plants (about 60 days old) covered with iron wire mesh ovipositioning cage and after 4 days the eggs were collected and sterilized. Obtained larvae were transferred to fresh diet and reared until pupation; healthy pupae were transferred to Petri dishes with filter paper. Pupae with disease symptoms were discarded and healthy pupae were placed in an emergence cage provided with moistened synthetic sponge at the bottom. The plants for the assessment insecticidal properties

were chosen on the basis of their resistance to insect attack, known poisonous properties and presence of odoriferous components. Selected plants were air-dried under shade at room temperature, dried materials were ground to powder in simple electric grindings mill, and 100 g sample of the ground plant material was taken for extraction. The sequential extraction of plant material was carried out in a soxhlet extractor using different solvents in a sequence on the basis of their polarity. The sequence of solvents followed like petroleum ether, chloroform, methanol and distilled water. The

extraction was done at 40 to 100°C for 20 to 24 h with each solvent accordingly and solvents were removed with the help of flash point evaporator. Used 10 mg of each fraction was diluted to 50 ml with analytic reagent (AR) grade acetone. The test solutions of required concentrations was prepared by diluting the required quantity of aforementioned stock solution with acetone and makeup volume of 100 ml in a volumetric flask. Efficacy of plant extracts on the ovipo-sitional behaviour, viability of eggs and incubation period were tested. Oviposition jars of 6 × 8” were used for the study purpose and each jar was provided with 2 cm thick layer of moist sand placed at the bottom. Sand layer was covered with a circular piece of paper and was divided into five sectors of equal areas and the inner sidewall as well. Each strip/sector was treated with different fractions of a particular plant product dissolved in acetone. However, one strip/sector was treated with acetone only and untreated check was also kept. Two pairs of newly emerged adults were released inside each jar and covered with muslin cloth. The

eggs from each strip and sector were kept in Petri dishes and the numbers of larvae hatched were recorded every day. Observations were also made for incubation period, viability and oviposition index. For ovicidal action against the eggs where the homogenous egg mass of one-two and three days age were selected and eggs masses were dipped in plant extract for few five seconds. Six concentrations were tested against eggs of different age groups and check (acetone treated) and untreated (control) was also run

simultaneously and average percentage of eggs hatched was recorded. Each treatment group had three replicates and observed percent ovicidal activity was corrected by using Abbott’s standard formula (Abbott, 1925). On emergence, moths were collected and released for egg-laying in the ovipositioning cage. The experiments were carried out in the laboratory at 27 ± 2°C and 65 ± 5% RH. The data pertaining to different experiments were subjected to statistical analysis by standard practices using completely randomized design (CRD). The sever root transformation was used in ovipositional be-

haviour experiments by adding a factor of 0.5; while in ovicidal and survival experiments, the data were transformed using angular transformation. The analysis was carried out by “micro-32” computer.

RESULTS Effects of Azadirachta indica A. juss leave extract Effect on oviposition Studies on the oviposition deterrency of extracts revealed that the number of eggs laid on untreated areas (control) were more than in all other treatments. The average number of eggs (101.66 and 77%) eggs were laid on control and check strips/sectors respectively, when compared to methanol 15, chloroform 21, petroleum ether 33 and distilled water 42% extracts at 5% concentration. The data showed order of effectiveness of various extracts as follows: methanol, extract, chloroform extract, petroleum ether extract and distilled water extracts at 10% concentration. Similar orders of effectiveness of extracts were found at 20% concentration. This clearly revealed that methanol extract was most effective and reduced the oviposition on treated strips/sector when compared to other extracts. The viability of eggs which were laid on the treated strips/sectors were effected slightly, lowest oviposition index (0.08) was recorded at 20% level of methanol extract, 10% (0.11) and 5% (0.14) concentrations of respective extracts. The oviposition index of chloroform extract treated strips/sector at 20, 10 and 5% concentration gave 0.12, 0.16 and 0.20 respec-tively. Average number of egg masses, three in number, on all the treated strips/sectors, while 4 to 5 days in control and check. It was also found that all the tested extract of A. indica did not affect the average hatching periods of eggs (Table 1). Effect on larval survival The data revealed that at 20% concentration, 24 h after treatment, methanol extract gave the lowest survival (55%) of larvae, which was better than chloroform extract (62%). The other tested extracts were not found to be significantly superior after 24 h exposure. The lower concentrations of extracts were not found to be effective. Chloroform methanol extracts at 20%concentration (55and 52.33 %) survival of larvae followed by petroleum ether (73%) and distilled water extract (75%). Similarly, survival of larvae (75%) was recorded at 10% concentration of water extract treated leaves, while it was 80% on chloroform extract treated leaves at 10% concentration. After 72 h exposure, chloroform and methanol extracts which proved to be significantly the best extracts amongst the tested extracts showed 52.33% at 20% concentration, where as petroleum ether and distilled water extracts stood moderate in their activity at 20% concentration (73 and 75%), at par with control and check. However, after 24, 48 and 72 h, factors “a” and “b” showed high significance (Table 2).

Mumtaz et al. 2647 Ovicidal action Observed one day old eggs that the treatments like, petroleum ether and chloroform extract at 10 and 20% concentrations were found to be significantly superior and gave (68, 68and 65, 65%) survival of eggs respectively. While other extracts were found to be least effective. For two days old eggs were comparatively more susceptible to petroleum ether and chloroform extracts then methanol and water extracts and for three day old eggs, the petroleum ether extract at 8, 10 and 20% concentrations were found to be the (60, 55 and 52%) survival of eggs, while the chloroform extract at the similar concentrations gave (75, 60 and 54%) survival of eggs respectively (Table 3). Effects of Callistemon citrinus D. C. leave extract Effect on oviposition In the experimental studies on the oviposition deterrency of various extracts, petroleum ether and chloroform extracts, it was observed that 5, 10 and 20% chloroform and petroleum ether leaves extracts were found to be significantly superior to control and gave 16, 14 and 10, and 17, 16 and 11% average number of eggs on the treated strips/sector respectively. The lowest oviposition index, 0.14, was recorded for chloroform ether extract treated strip/sector at 20% concentration, while the highest index, 0.73%, was recorded for the check treatment at a similar concentration, clearly indicating that the check had the least effect on the oviposition behavior. The average number of egg masses little bid varied, and average hatching period of eggs were not affected at all (Table 4). Effect on larval survival Results showed that 24 h exposure of different treated leaves revealed that petroleum ether extract of leaves gave maximum reduction in the survival of 3 to 4 days old larvae of C. auricilius (55%) and was better than metha-nol, chloroform and distilled water extracts. However, the survival of larvae was 100% in control and check.

The survival of larvae decreased after 48 and 72 h exposure (48.66 and 48.66%) to petroleum ether leaves extract, also observed that only higher concentration of extract had slight effect on larval survival, while lower concentration of extracts were not effective of larvae at 24, 48 and 72 h after exposure (Table 5). Ovicidal action Tested leaves extract had slight variation in their efficacy

2648 J. Med. Plants Res. Table 1. Number of eggs laid by Chilo auricilius and their viability on treated strips/sectors of Azadirachta indica.

Treatments plant extracts

Average number of eggs laid by female

Average number of eggs hatched Oviposition index

Average number of

egg mass

Average hatching period (in days)

Concentration (%) Concentration (%) Concentration (%) Concentration

(%) Concentration (%)

5 10 20 Mean 5 10 20 Mean 5 10 20 5 10 20 5 10 20

Petroleum

ether extract

33.00

(5.78)

28.00

(5.33)

21.00

(4.63)

27.33

(5.25)

33.00

(5.78)

28.00

(5.33)

21.00

(4.63)

27.33

(5.25)

0.35

(0.92)

0.23

(0.85)

0.18

(0.82)

3 3 3 3 3 3

Chloroform

extract

21.00

(4.63)

17.00

(4.17)

16.00

(4.05)

18.00

(4.28)

21.00

(4.63)

17.00

(4.17)

16.00

(4.05)

18.00

(4.28)

0.20

(0.83)

0.16

(0.81)

0.12

(0.78)

3 3 3 3 3 3

Methanol

extract

15.00

(3.92)

10.00

(3.23)

12.00

(3.52)

12.33

(3.56)

15.00

(3.92)

10.00

(3.23)

12.00

(3.52)

12.33

(3.56)

0.14

(0.79)

0.11

(0.78)

0.08

(0.73)

3 3 3 3 3 3

Distilled

water extract

42.00

(6.50)

36.00

(6.04)

33.00

(5.78)

37.00

(6.10)

42.00

(6.50)

36.00

(6.04)

33.00

(5.78)

37.00

(6.10)

0.40

(0.94)

0.31

(0.89)

0.25

(0.86)

3 3 3 3 3 3

Acetone as check

77.00

(8.80)

68.00

(8.27)

67.00

(8.21)

70.66

(8.43)

77.00

(8.80)

68.00

(8.27)

67.00

(8.21)

70.66

(8.43)

0.75

(1.11)

0.55

(1.02)

0.51

(1.00)

4 4 4 3 3 3

Control 101.66

(10.10)

117.66

(10.87)

128.00

(11.33)

115.77

(10.77)

101.66

(10.10)

117.66

(10.87)

128.00

(11.33)

115.77

(10.77)

- - - 5 5 5 3 3 3

Factor Sig. SEm ± CD at 5% Sig. SEm ± CD at 5%

A ** (**) 1.03 (0.84) 2.96 (0.24) ** (**) 1.03 (0.84) 2.96 (0.24)

B ns (**) 0.73 (0.59) 2.09 (0.17) ns (**) 0.73 (0.59) 2.09 (0.17)

A B ** (**) 1.78 (0.14) 5.12 (0.41) ** (**) 1.78 (0.14) 5.12 (0.41)

Data in parentheses represents square root transformation; A = Concentration, B = Solvent.

against the eggs of C. auricilius, that is, higher susceptibility of older eggs (two and three days old), when compared to newly deposited eggs (one day old). The chloroform leaves extract gave the lowest survival (61%) of three days old eggs

at 20% concentration, and ranked better than methanol leaves extract (69%) at 20% concentration; and observation was recorded after 24 h. The 10% concentration of chloroform and methanol leaves extracts were also found to be

significantly superior over control on three days old eggs. For three days old eggs, 20 and 10% concentrations of methanol leaves extract proved more or less equally effective.

The 10% concentration of chloroform leaves

Mumtaz et al. 2649 Table 2. Survival of Chilo auricilius larvae on different leaves extracts of Azadirachta indica (feeding method).

Treatments plants extract concentration (%)

(%) Survival of larvae

After 24 h exposure After 48 h exposure After 72 h exposure

Petroleum ether

extract

Chloroform extract

Methanol extract

Distilled

water

extract

Mean Petroleum

Ether extract

Chloroform Extract

Methanol Extract

Distilled

water

extract

Mean

Petroleum

Ether

extract

Chloroform Extract

Methanol Extract

Distilled

water

extract

Mean

2 92.00

(73.65)

100.00 (90.00)

98.00

(83.35)

100.00

(90.00)

97.50

(84.25)

85.00

(67.39)

95.66

(80.21)

93.00

(74.89)

94.00

(75.90)

91.91

(74.60)

85.00

(67.39)

95.00

(79.40)

88.00

(69.85)

94.00

(81.63)

90.50

(74.57)

4 92.00

(73.65)

92.00

(73.65)

92.00

(73.65)

97.00

(81.84)

93.25

(75.704)

85.00

(67.392)

85.00

(67.39)

93.00

(74.89)

91.00

(72.59)

88.50

(70.57)

85.00

(67.39)

85.00

(67.39)

84.00

(66.49)

90.00

(71.62)

86.00

(68.22)

8 85.00

(67.39)

85.00

(67.39)

83.00

(68.26)

92.00

(73.65)

86.25

(69.17)

82.00

(64.96)

83.00

(68.26)

75.00

(60.02)

83.00

(65.71)

80.75

(64.74)

82.00

(64.91)

80.00

(63.44)

73.00

(58.72)

81.00

(64.16)

79.00

(62.81)

10 82.00

(65.03)

81.00

(64.17)

72.00

(58.18)

85.00

(67.39)

80.00

(63.69)

73.00

(58.72)

80.00

(63.63)

65.00

(53.76)

75.00

(60.03)

73.25

(59.03)

75.00

(60.03)

73.00

(58.72)

65.00

(53.76)

75.00

(60.03)

72.00

(58.13)

20 77.00

(61.42)

62.00

(51.96)

55.00

(47.88)

80.00

(63.45)

68.50

(56.18)

73.00

(58.72)

55.00

(47.87)

52.33

(46.34)

75.00

(60.03)

63.83

(53.24)

73.00

(58.72)

52.33

(46.34)

52.33

(46.34)

75.00

(60.03)

63.16

(52.85)

Acetone as check

100.00

(90.00)

97.00

(81.84)

97.00

(81.84)

100.00

(90.00)

98.50

(85.92)

100.00

(90.00)

97.00

(84.18)

97.00

(84.18)

100.00

(90.00)

98.50

(87.09)

100.00

(90.00)

97.00

(84.18)

97.00

(84.18)

97.00

(84.18)

97.75

(85.63)

Control 100.00

(90.00)

97.00

(81.84)

100.00

(90.00)

100.00

(90.00)

99.25

(87.96)

100.00

(90.00)

98.00

(83.44)

100.00

(90.00)

100.00

(90.00)

99.50

(88.36)

100.00

(90.00)

100.00

(90.00)

100.00

(90.00)

100.00

(90.00)

100.00

(90.00)

Factor Sig. SEm ± CD at 5% Sig. SEm ± CD at 5% Sig. SEm ± CD at 5%

A ** (**) 1.09 (1.03) 3.09 (2.92) ** (ns) 1.19 (1.12) 3.37 (3.18) ** (**) 1.02 (1.14) 2.90 (3.25)

B ** (**) 1.44 (1.36) 4.08 (3.86) ** (**) 1.57 (1.48) 4.46 (4.21) ** (**) 1.35 (1.51) 3.83 (4.30)

A B ** (**) 2.88 (2.72) 8.17 (7.72) ** (*) 3.15 (2.97) 8.93 (8.43) ** (ns) 2.70 (3.03) 7.67 (8.60)

Data in parentheses represents angular transformation; A = solvent, B = concentration.

extract also minimized the survival of eggs (64%) significantly than rest of the other treatments. The survival of three days old eggs was 89% in check and slightly lower than that recorded in untreated eggs (94%). Factor b of leaves extracts was highly significant at one, two and three days old

eggs (Table 6). DISCUSSION A. indica extracts on the eggs of Dysdercus

cingulatus Fabricius, highest survival (12.25%) at 0.005% concentration of green neem seed coat and least survival of eggs were observed at 0.005% neem seed kernel extracts after one week (Tanu et al., 2010). Plant products were used as insecticides for the control of Scirpophaga

2650 J. Med. Plants Res. Table 3. Ovicidal action of leaves extracts of Azadirachta indica against Chilo auricilius.

Treatments plants extract concentration

(%)

(%) Survival of eggs

One day old eggs Two days old eggs Three days old eggs

Petroleum ether

extract

Chloroform extract

Methanol extract

Distilled

water

extract

Mean Petroleum

ether extract

Chloroform extract

Methanol extract

Distilled

water

extract

Mean

Petroleum ether

extract

Chloroform extract

Methanol extract

Distilled

water

extract

Mean

2 75.00

(60.03)

84.00

(66.63)

80.00

(63.63)

85.00

(67.70)

81.00

(64.50)

74.00

(59.35)

83.00

(65.69)

78.00

(62.05)

82.00

(64.91)

79.25

(63.00)

70.00

(56.81)

80.00

(63.44)

70.00

(56.81)

80.00

(63.45)

75.00

(60.13)

4 75.00

(60.03)

80.00

(63.63)

80.00

(63.63)

83.00

(65.69)

79.50

(63.24)

73.00

(58.71)

79.66

(63.20)

78.00

(62.054)

80.00

(63.639)

77.66

(61.90)

65.00

(53.76)

75.00

(60.03)

70.00

(56.81)

75.00

(60.03)

71.25

(57.66)

8 75.00

(60.03)

77.00

(61.35)

80.00

(63.63)

80.00

(63.63)

78.00

(62.16)

65.00

(53.76)

77.33

(61.61)

74.00

(59.35)

80.00

(63.63)

74.08

(59.58)

60.00

(50.77)

75.00

(60.03)

65.00

(53.76)

75.00

(60.03)

68.75

(56.15)

10 68.00

(55.76)

68.00

(55.76)

78.00

(62.05)

80.00

(63.63)

73.50

(59.30)

65.00

(53.76)

65.00

(53.76)

74.00

(59.35)

75.00

(60.03)

69.75

(56.72)

55.00

(47.87)

60.00

(50.77)

60.00

(50.77)

70.00

(56.81)

61.25

(51.55)

20 65.00

(53.76)

65.00

(53.76)

75.00

(60.03)

75.00

(60.03)

70.00

(56.89)

58.00

(49.60)

59.00

(50.18)

70.00

(56.81)

73.00

(58.71)

65.00

(53.82)

52.00

(46.14)

54.00

(47.29)

58.00

(49.60)

65.00

(53.76)

57.25

(49.20)

Acetone as check

88.00

(69.85)

88.00

(69.85)

88.00

(69.85)

88.00

(69.85)

88.00

(69.85)

88.00

(69.85)

88.00

(69.85)

88.00

(69.85)

88.00

(69.85)

88.00

(69.85)

93.00

(74.92)

93.00

(74.92)

93.00

(74.92)

93.00

(74.92)

93.00

(74.92)

Control 88.00

(69.85)

88.00

(69.85)

88.00 (69.85)

88.00

(69.85)

88.00

(69.85)

90.00

(71.91)

90.00

(71.91)

90.00

(71.91)

90.00

(71.91)

90.00

(71.91)

94.00

(75.90)

94.00

(75.90)

94.00

(75.90)

94.00

(75.90)

94.00

(75.90)

Factor Sig. SEm ± CD at 5% Sig. SEm ± CD at 5% Sig. SEm ± CD at 5%

A ** (**) 1.24 (0.86) 3.52 (2.43) ** (**) 0.85 (0.64) 2.42 (1.82) ** (**) 0.70 (0.52) 2.00 (1.47)

B ** (**) 1.64 (1.13) 4.66 (3.22) ** (**) 1.13 (0.85) 3.20 (2.41) ** (**) 0.93 (0.69) 2.65 (1.95)

A B ns (ns) 3.29 (2.27) 9.33 (6.44) ** (**) 2.26 (1.70) 6.40 (4.82) ** (**) 1.87 (1.38) 5.30 (3.91)

Data in parentheses represents angular transformation; A = Solution, B = Concentration.

excerptalis (Pandey et al., 1998). The lowest number of pests (6.35 and 6.17%) was observed following treatments with Eucalyptus rostrata (3%) compared to the untreated control (16.88 and

15.77%) respectively followed by A. indica and E. rostrata (2% solution). Effects of water extracts of four plant leaves in insect pest control with various concentrations of the extracts (8.5, 1, 2, 4 and

6%) were used against last instar larvae of the groundnut pest Spodoptera litura. Calotropis gigantean was found to be the most toxic plant product followed by Vitex negundo, A. indica and

Mumtaz et al. 2651 Table 4. Number of eggs laid by Chilo auricilius and their viability on treated strips/sectors of Callistemon citrinus leaves extracts (choice method).

Treatments

plant extracts

Average number of eggs laid by female

Average number of eggs hatched

Oviposition index

Average number of

egg mass

Average hatching period

(in days)

Extract concentration (%) Extract concentration (%) Extract concentration (%) Extract concentration (%) Extract concentration (%)

5 10 20 Mean 5 10 20 Mean 5 10 20 5 10 20 5 10 20

Petroleum

ether extract

17.00

(4.17)

16.00

(4.05)

11.00

(3.38)

14.66

(3.86)

13.00

(3.66)

11.00

(3.38)

9.00

(3.07)

11.00

(3.37)

0.27

(0.87)

0.24

(0.86)

0.18

(0.82)

2.0 2.0 2.0 3.0 3.0 3.0

Chloroform

extract

16.00

(4.05)

14.00

(3.80)

10.00

(3.23)

13.33

(3.69)

8.00

(2.90)

7.00

(2.73)

6.00

(2.53)

7.00

(2.72)

0.24

(0.86)

0.19

(0.83)

0.14

(0.79)

3.0 3.0 3.0 3.0 3.0 3.0

Methanol

extract

32.00

(5.68)

28.00

(5.33)

23.00

(4.84)

27.66

(5.28)

28.00

(5.33)

20.00

(4.52)

16.00

(4.05)

21.33

(4.63)

0.41

(0.95)

0.38

(0.93)

0.26

(0.87)

3.0 3.0 4.0 3.0 4.0 4.0

Distilled

water extract

47.00

(6.89)

45.00

(6.73)

38.00

(6.20)

43.33

(6.61)

40.00

(6.36)

38.00

(6.19)

31.00

(5.61)

36.33

(6.05)

0.61

(1.05)

0.58

(1.03)

0.45

(0.97)

3.0 4.0 4.0 3.0 4.0 4.0

Acetone as check

66.00

(8.15)

58.00

(7.64)

55.00

(7.44)

59.66

(7.74)

58.00

(7.64)

50.00

(7.08)

47.00

(6.88)

51.66

(7.20)

0.87

(1.17)

0.43

(0.96)

0.73

(1.10)

3.0 4.0 4.0 3.0 4.0 4.0

Control 75.00

(8.68)

77.00

(8.80)

80.00

(8.96)

77.33

(8.81)

68.00

(8.27)

72.00

(8.51)

72.00

(8.51)

70.66

(8.43)

- - - 3.0 4.0 6.0 4.0 4.0 4.0

Factor Sig. SEm ± CD at 5% Sig. SEm ± CD at 5%

A ** (**) 1.15 (0.96) 3.30 (0.27) ** (**) 1.03 (0.87) 2.95 (0.25)

B ** (**) 0.81 (0.68) 2.33 (0.19) ** (**) 0.72 (0.61) 2.08 (0.17)

A B ** (ns) 1.99 (0.16) 5.72 (0.47) ** (**) 1.78 (0.15) 5.11 (0.43)

Data in parentheses represents square root transformation; A = Concentration, B = Solvent

Pinus glabra (Pometia pinnata) (Sahayraj, 1998). Extracts of A. indica, Annona squamosa, Momor-dica charantina, Lantana camara, P. glabra, Diospyros cordifolia and Garlic against Amsacta morei, five percent suspension sprays of A. indica seed and leaf, custard apple seed and leaf, bitter gourd seed, Lantana leaf, Kanzi seed, dhaki seed and garlic bulb have been found considerably effective against 2nd instar larvae of A. moorei. However, A. indica seed was found significantly

superior to the rest of the treatments by giving 92.22% mortality, followed by A. indica leaf extract, which gave 72.22% larval mortality. The custard seed extract gave 36.67% mortality where the rest of the extracts gave larval mortality in the range of 46.67 to 54.44% (Patel et al., 1993). Toxic effects of plant extracts on eggs of Chilo partellus, a pest of maize, several semi-solid crude plant metabolites against 3 day old eggs of C. partellus. Petroleum ether fractions of A.

squamosa and A. indica were the most toxic and these completely inhibited hatching (Bhatnagar, 1995). The other tested extracts also proved to be effective but their effects were too slow (Singh et al., 1997). However, the factors a, b and their interaction showed high significance at one, two and three days old eggs. Extract of A. indica leaf mainly contains meliantriol, Azadirachtin and Salanin. The leaves of A. indica also contain nim-bin, nimbinin, 6- desacetylnimbinene, nimbandiol,

2652 J. Med. Plants Res. Table 5. Survival of Chilo auricilius larvae on different leaves extracts Callistemon citrinus (feeding method)

Treatments

plants extract

concentration (%)

(%) Survival of Larvae

After 24 h exposure After 48 h exposure After 72 h exposure

Petroleum

Ether

extract

Chloroform

Extract

Methanol

Extract

Distilled

water

extract

Mean

Petroleum

Ether

extract

Chloroform

Extract

Methanol

Extract

Distilled

water

extract

Mean

Petroleu

m Ether

extract

Chloro

form

Extract

Methanol

Extract

Distilled

water

extract

Mean

2 85.00

(67.39)

92.00

(73.65)

100.00

(90.00)

100.00

(90.00)

94.25

(80.26)

85.00

(67.39)

88.00

(69.85)

95.00

(79.40)

94.00

(81.63)

90.50

(74.57)

85.00

(67.39)

88.00

(69.85)

95.00

(79.40)

94.00

(81.63)

90.50

(74.57)

4 80.66

(64.06)

92.00

(73.86)

100.00

(90.00)

100.00

(90.00)

93.16

(79.48)

80.00

(63.63)

80.00

(63.44)

92.00

(73.82)

94.00

(81.63)

86.50

(70.63)

80.00

(63.63)

80.00

(63.44)

92.00

(73.82)

94.00

(81.63)

86.50

(70.63)

8 62.00

(51.95)

87.00

(69.09)

97.00

(80.27)

97.00

(80.27)

85.75

(70.39)

62.00

(51.95)

80.00

(63.44)

90.00

(71.62)

85.00

(67.39)

79.25

(63.60)

62.00

(51.95)

80.00

(63.44)

90.00

(71.62)

85.00

(67.39)

79.25

(63.60)

10 61.00

(51.35)

75.00

(60.03)

83.00

(65.69)

81.33

(66.92)

75.08

(61.00)

60.00

(50.77)

75.00

(60.03)

78.00

(62.05)

74.00

(59.35)

71.75

(58.05)

60.00

(50.77)

75.00

(60.03)

78.00

(62.05)

74.00

(59.35)

71.75

(58.05)

20 55.00

(47.89)

72.00

(58.08)

62.00

(51.96)

74.00

(59.46)

63.25

(52.85)

48.66

(44.23)

70.00

(56.95)

60.00

(50.70)

74.00

(59.35)

63.16

(52.82)

48.66

(44.23)

70.00

(56.95)

60.00

(50.77)

74.00

(59.35)

63.16

(52.82)

Acetone as check 100.00

(90.00)

100.00

(90.00)

100.00

(90.00)

100.00

(90.00)

100.00

(90.00)

97.00

(84.18)

94.00

(81.63)

97.00

(84.18)

97.00

(84.18)

96.25

(83.54)

97.00

(84.18)

94.00

(81.63)

97.00

(84.18)

97.00

(84.18)

96.25

(83.54)

Control 100.00

(90.00)

100.00

(90.00)

100.00

(90.00)

100.00

(90.00)

100.00

(90.00)

97.00

(84.18)

97.00

(84.18)

97.00

(84.18)

97.00

(84.18)

97.00

(84.18)

97.00

(84.18)

97.00

(84.18)

97.00

(84.18)

97.00

(84.18)

97.00

(84.18)

Factor Sig. SEm ± CD at 5% Sig. SEm ± CD at 5% Sig. SEm ± CD at 5%

A ** (**) 1.41 (0.96) 3.99 (2.74) ** (**) 1.19 (1.63) 3.37 (4.63) ** (**) 1.19 (1.63) 3.37 (4.63)

B ** (**) 1.86 (1.28) 5.28 (3.62) ** (**) 1.57 (2.16) 4.46 (6.13) ** (**) 1.57 (2.16) 4.46 (6.13)

A B ** (**) 3.73 (2.56) 10.57 (7.25) ns (ns) 3.15 (4.33) 8.93 (12.26) ns (ns) 3.15 (4.33) 8.93 (12.26)

Data in parentheses represents angular transformation. A = Solvent, B = Concentration.

Mumtaz et al. 2653 Table 6. Ovicidal action of Leaves extracts of Callistemon citrinus against Chilo auricilius.

Treatments plants extract concentration (%)

(%) Survival of eggs

One day old eggs

Two days old eggs

Three days old eggs

Petroleum ether

extract

Chloroform extract

Methanol extract

Distilled water

extract

Mean

Petroleum ether

extract

Chloroform extract

Methanol extract

Distilled water

extract

Mean Petroleum

ether extract

Chloroform extract

Methanol extract

Distilled water

extract

Mean

2 81.00

(64.16)

81.00

(64.16)

83.00

(68.26)

84.00

(66.63)

82.25

(65.80)

75.00

(60.43)

80.00

(63.63)

80.00

(63.63)

81.0

(64.16)

79.00

(62.96)

76.00

(60.92)

75.00

(60.03)

75.00

(60.43)

79.00

(62.96)

76.25

(61.08)

4 80.00

(63.63)

78.00

(62.05)

76.00

(60.67)

80.00

(63.63)

78.50

(62.49)

75.00

(60.03)

75.00

(60.03)

74.00

(59.35)

80.00

(63.63)

76.25

(60.89)

75.00

(60.03)

72.00

(58.18)

74.00

(59.69)

79.00

(62.96)

75.00

(60.22)

8 80.00

(63.63)

74.00

(59.35)

74.00

(59.35)

76.00

(60.67)

76.00

(60.75)

74.00

(59.35)

74.00

(59.35)

74.00

(59.35)

75.00

(60.03)

75.25

(60.25)

74.00

(59.69)

70.00

(56.95)

73.00

(58.71)

74.00

(59.35)

72.75

(58.67)

10 75.00

(60.03)

74.00

(59.35)

73.00

(58.71)

76.00

(60.67)

74.50

(59.69)

75.66

(60.45)

71.00

(57.42)

73.00

(58.71)

76.00

(60.92)

73.91

(59.37)

73.00

(58.71)

64.00

(53.13)

70.00

(56.95)

74.00

(59.35)

70.25

(57.03)

20 75.00

(60.03)

73.00

(58.71)

72.00

(58.18)

75.00

(60.03)

73.50

(59.05)

70.00

(56.95)

68.00

(55.76)

72.00

(58.18)

74.00

(59.69)

71.00

(57.65)

70.00

(56.95)

61.00

(51.36)

69.00

(56.18)

73.00

(58.71)

68.25

(55.80)

Acetone as

check

86.00

(68.05)

86.00

(68.05)

86.00

(68.05)

86.00

(68.05)

86.00

(68.05)

87.00

(69.09)

87.00

(69.09)

87.00

(69.09)

87.00

(69.09)

87.00

(69.09)

89.00

(70.66)

89.00

(70.66)

89.00

(70.66)

89.00

(70.66)

89.00

(70.66)

Control 87.00

(69.09)

87.00

(69.09)

87.00

(69.09)

87.00

(69.09)

87.00

(69.09)

87.00

(69.09)

90.00

(71.91)

90.00

(71.91)

90.00

(71.91)

89.25

(71.20)

94.00

(75.90)

94.00

(75.90)

94.00

(75.90)

94.00

(75.90)

94.00

(75.90)

Factor Sig. SEm ± CD at 5% Sig. SEm ± CD at 5% Sig. SEm ± CD at 5%

A ns (ns) 1.13 (0.87) 3.20 (2.46) ns (ns) 1.36 (0.95) 3.87 (2.69) ns (ns) 1.34 (0.88) 3.80 (2.51)

B ** (**) 1.49 (1.15) 4.23 (3.26) ** (**) 1.80 (1.25) 5.11 (3.56) ** (**) 1.77 (1.17) 5.03 (3.32)

A B ns (ns) 2.99 (2.30) 8.47 (6.52) ns (ns) 3.61 (2.51) 10.23 (7.12) ns (ns) 3.55 (2.34) 10.07 (6.64)

Data in parentheses represents angular transformation; A = Solution, B = Concentration.

nimbolide and qnerectin. Azadirachtin isolated neem leaves and seeds have repellent effect. It is possible that repellent activity of methanol and chloroform extracts to the female moth may be due to the high level of Azadirachtin and other

toxic compounds. It was observed that deterrent effect on egg laying gradually increased with increased in the concentration of extracts from 5 to 20%, and this can be attributed to the presence of increased level of principal components of

Azadirachtin at each level (Warthren, 1979). Adverse effects on oviposition by the application of A. indica products had been reported in case of Spodoptera litura, Nilaparvata lugens, E. variastis (Steets and Schmutterer, 1975; Joshi and

2654 J. Med. Plants Res. Sitaramaiah, 1979; Saxena, et al., 1979). Ether and ethanol extracts of various part of A. indica against cotton bug, Dysdercus cingulattus Fab. Azadirachtin, a strong feeding deterrent for Spodoptera gregaria (forsk) is isolated from the A. indica leaves and seeds (Singh et al., 1997). A. indica leaves compound had been considered to be potential value against various crop pests (Gill and Lewis, 1971). A. indica seed kernel extracts was effective against sorghum insect pests, such as maize stem borer, C. partellus (Swinhoe), army worm Mythimna separata (Walker) and the ear head bug. Calocoris angustatus Let. by the application of extract, there was an increased in yield of sorghum by 25-30% (Sankaran et al., 1986). The juvenile hormone effects on C. partellus (larvae) (Gujar and Mehrotra, 1983). Evaluated the efficacy of plant products as insecticide for the control of Scirpophaga excerptalis and observed the lowest number of pests 6.35 and 6.17% of following treatment with Eucalyptus rostrata (3%) compared to the untreated control 16.88 and 15.77%, respectively, followed by A. indica and E. rostrata (2% solution) (Pandey et al.,1998). However, A. indica leaf and seed extracts were affected the oviposition activity of many other insects (Hellpap and Mercado, 1986).

Data showed that three days old eggs of C. auricilius was the most susceptible, when compared to two days and one day old eggs. Petroleum ether extract was found to be the most effective against three days old eggs. Toxic effect of plant extracts on eggs of C. partellus, petroleum ether fractions of Annona squamosa and A. indica were the most toxic and these completely inhibited hatching, and survival data showed lowest survival (55%) at 20% concentration of methanol extract followed by 62% of chloroform extract treated leaves at 24 h after treatment. The higher toxicity of methanol and chloroform extracts was reduced and it may be due to the presence of high amount of Azadirachtin in the extracts (Bhatnagar, 1995). Nishinda leaf powder effectively prevented oviposition by the corn weevil (Bhuiyan and Quiniones, 1990). A mixture of food with pithraj leaf, bark and seed powder reduced the oviposition rates of the pulse beetle (Talukder and Howse, 1994). Extracts from nine plant materials which act as oviposition deterrents against C. maculatus showed that pulse treated with Rhazya stricta leaves A. indica seeds, Heliotropium bacciferum aerial parts and citrus peels acted as the highest ovipositional deterrents (Elhag, 2000). Dry ground leaf of Chenopodium ambrosioides inhibited F1 progeny production and adult emergence of the C. chinensis and C. maculates (Tapondjou et al., 2002). Adverse morpho-genetic and gonadotrophic effects of Australian bottle brush, C. lanceolatus had been reported in case of Dysedurcus koengii Fabr., when extract were applied topically on final instar nymph (Katiyar and Srivastava, 1982). Chemically, the bottle brush plant have high amount of alkaloids, which might be responsible for deterrent, ovicidal and antifeedant activity against C. auricilius. Among the various extracts of this plant, it was

recorded that chloroform leaves extract had affected the oviposition, while survival activity of larvae was affected by petroleum ether and methanol leaves extract. The chloroform leaves extract showed ovicidal activity, while the fruits extracts remained ineffective against C. auricilius. Carbon tetrachloride extracts of leaves’ extract of C. citrinus showed significant antimicrobial activity (Mohammad et al., 2010).

The findings of the present investigations indicate that botanical derivatives might be useful as insect control agents. A. indica and C. citrinus extracts tested were effective to some degree in reducing the ovipositional preferences, and increasing the inhibition rates of the larval survival and ovicidal action. A. indica leaves extract was comparatively effective than C. cirtinus among leaves extract against C. auricius. As per low concen-trations of these two plants extract were less effective against the target pest. Using of plants materials are more economic in extract preparations and more effective in toxicity and slow down the development of insect resistance (Dadang et al., 2008). Moreover, to minimize the severe damage caused by insect pests, the traditional use of plant products proved to be highly effective against insects. Application of plant extracts an inexpensive and effective technique and its easy adaptability will give additional advantages leading to acceptances of this technology by farmers. A study to improve the effectiveness of botanical derivatives as insecticides will benefit agricultural sectors of developing countries, as these substance are not only of low cost, but also have less environmental impact in term of insecticidal hazard.

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