SHORT COMMUNICATION

Preliminary Assessment of Selected Botanicals in the Controlof Tetranychus neocaledonicus Andre (Acari: Tetranychidae)

Indranil Roy • Gautam Aditya • Goutam K. Saha

Received: 5 November 2010 / Revised: 7 July 2011 / Accepted: 24 October 2011 / Published online: 10 November 2011

� Zoological Society, Kolkata, India 2011

Abstract The mites Tetranychus neocaledonicus Andre,

1933 (Acari: Tetranychidae) are pest of different medicinal

plants. In view of biological regulation of this mite, an

evaluation of certain botanicals in regulation of the mite

population was made under laboratory conditions. In the

present study recorded 72 h after the exposure of T. neo-

caledonicus on the leaf of Withania somnifera to the 3%

aqueous dose of plant extracts, 61.1% mortality was seen

against Eupatorium triplinerve, 75% mortality was recor-

ded against Cassia alata and 82.2% was found against

Ocimum tenuiflorum. On the other hand, 3% methanolic

dose of E. triplinerve and O. tenuiflorum extracts were

responsible for 74.4 and 93.3% mortality of mites,

respectively, whereas highest mortality (97%) was recor-

ded by 3 per cent methanolic dose of C. alata.

Keywords Biopesticide � Mites � Pest � Tetranychus

neocaledonicus

Introduction

The extracts of different angiosperm plants have been

screened for arthropod vector and pest control worldwide

(Arnason et al. 1989; Sarmah et al. 1999; Macchioni et al.

2004; Rahman et al. 2005; Jbilou et al. 2006; Sarmah et al.

2009). Apart from the origin of the resistant strains of

insect and mite pests, use of plant extracts are beneficial

considering the environmental ill effects and cost of the

pesticides, many of them are known to affect crop, orna-

mental and medicinal plants (Sadana 1985; Chhillar et al.

2007). Many of these species are vulnerable to the extracts

of neem (Sundaram et al. 1995; Ismail et al. 2007; Venzon

et al. 2008), Acorus calamus (L.), Xanthium strumarium

(L.), Polygonum hydropiper (L.) and Clerodendron infor-

tunatum (Gaertn) (Sarmah et al. 2009) signifying that plant

extracts can be a viable alternative in regulation of popu-

lation of mite pest in varied conditions.

The spider mite Tetranychus neocaledonicus Andre

(Acari: Tetranychidae) is a common phytophagous mite

associated with varieties of crop (Ehara and Yamaguchi

2001), medicinal and ornamental plants (Chhillar et al.

2007; Gupta 2005). It is widely distributed and in certain

instances causes heavy damage to crops (Chhillar et al.

2007). The colonies of T. neocaledonicus are formed on

the underside of the leaf, protected by a screen of silk

threads secreted by the females (Sadana 1985; Chhillar

et al. 2007). All stages (larvae, nymphs and adults) feed

on plant sap causing foliar damage. Feeding leads to the

disappearance of chloroplasts making tiny white spots on

the upper surface of the leaf. To combat the damage by

T. neocaledonicus on economically important plants,

chemical control may provide faster results with a pos-

sibility of backlash in terms of environmental degrada-

tion. Therefore, using fungi and plant products can be an

I. Roy � G. Aditya � G. K. Saha (&)

Department of Zoology, University of Calcutta, 35 Ballygunge

Circular Road, Calcutta 700019, India

e-mail: gkszoo@gmail.com; gkszoo@rediffmail.com

I. Roy

e-mail: indranilzoology@gmail.com

G. Aditya

e-mail: gautamaditya2001@gmail.com

G. Aditya

Department of Zoology, University of Burdwan, Golapbag,

Burdwan 713104, India

123

Proc Zool Soc (July-Dec 2011) 64(2):124–127

DOI 10.1007/s12595-011-0012-1

TH

EZ

O

OLOGICAL SOC

IET

YKO LK ATA

alternative in management of mite population. In the

present study a preliminary assessment of extracts of three

indigenous plant species has been evaluated in regulation

of mite population. The results are expected to highlight

the importance of these plants as a source of biopesticides

in regulating pest mites.

Materials and Methods

Leaf extracts were prepared from three widely used

medicinal plants viz. Eupatorium triplinerve Vahl.

(Family: Asteraceae) (common name: Ayapan), Cassia

alata Linn. (Family: Caesalpiniaceae) (common name:

Dadmari) and Ocimum tenuiflorum Linn. (Family: Lamia-

ceae) (common name: Krishnatulsi). An entomopathogenic

fungus viz. Paecilomyces fumosoroseus (1.15% WP)

[T. Stanes & Company Ltd., Coimbatore, India] and Neem

oil (Azadirachtin 0.03% EC) [T. Stanes & Company Ltd.,

Coimbatore, India] were also included to evaluate their

efficacy towards mortality of T. neocaledonicus Andre

under laboratory condition.

Preparation of Plant Extracts

Both methanolic and aqueous plant extract were prepared

following Macchioni et al. (2004). Leaves of three plants

were collected from Medicinal Plant Garden of Institute of

Agriculture and The Experimental Garden, Department of

Botany, University of Calcutta, Ballygunge, Kolkata.

Leaves of the plants were washed in tap and distilled water

followed by shade-dry for 2 weeks. The crispy leaves were

crushed in electric grinder. The crushed leaves were suc-

cessively extracted with methanol and water separately by

using Soxhlet apparatus for 72 h. The dried extract was

weighed and measured amount was dissolved in requisite

quantity of methanol and water for desired concentration.

In the present experiment 1, 2, 3 g crude extract was dis-

solved separately in 100 ml methanol and water for getting

1, 2 and 3% concentration.

Experimental Methods

The fresh leaves of Withania somnifera were dipped in the

plant extracts and the solution of biopesticides for 15 min

(Helle and Overmeer 1985) after which it was allowed to

dry for 20 min. Then the leaves were kept on a wet cotton

pad in a Petri dish (10 cm diameter and 1.5 cm in height).

The Petri dishes were placed under a cover of glass-wood

frame of appropriate size to prevent disturbance due to air

circulation in the laboratory at room temperature

(25–28�C) and[80% relative humidity. In case of controls,

the leaves were dipped in methanol and distilled water

respectively and allowed to dry and placed in Petri dish

in similar manner. Ten laboratory-reared adult female

T. neocaledonicus were released to each Petri dish. Nine

replicates were used per dose per plant extract. Three doses

for both aqueous and methanolic crude plant extracts

ranging between 1 and 3% (v/v) were put in use with

reference to the standard fungal and neem oil biopesticides

and the controls with equal number of replicates as treat-

ment. Individual Petri dish was examined under a stereo

binocular after 24, 48 and 72 h for counting the mortality.

The experiment was conducted twice and the data on

mortality, recorded at different intervals, were subjected to

ANOVA (Zar 1999) to infer about the differences among

the treatments.

Results

Both the aqueous and methanolic extracts of three plants

E. triplinerve, C. alata and O. tenuiflorum were found to

affect the survival rate of the mite, T. neocaledonicus on

leaf of W. somnifera, depending on the dose and the

period of exposure. The relative proportion of mortality

was found higher in methanolic extracts as compared to

aqueous extracts. The commercially formulated fungal

and neem oil resulted in higher mortality than the plant

extract but difference appeared insignificant. Aqueous and

methanolic extracts of the plants were found to be

effective only at a higher dose than those of commercial

ones after 72 h (Fig. 1). Exposure of T. neocaledonicus

on the leaf of W. somnifera to 3% aqueous dose of plant

extracts, 61.1% mortality was seen against E. triplinerve,

75% percent mortality was recorded against C. alata,

whereas 82.2% mortality was found against O. tenuiflo-

rum after the same period. On the contrary, 74.4 and

93.3% mortality of mites were observed with the use of

3% methanolic dose of E. triplinerve and O. tenuiflorum

extracts, highest mortality being 97% was recorded by 3%

methanolic dose of C. alata. No mortality was recorded in

the control set-ups. The results of ANOVA (Table 1)

indicate that the effectiveness of the methanolic extracts

differed significantly with the aqueous extracts. The

effectiveness of the extracts varied with the plant species

also, with C. alata exhibiting best results followed by

O. tenuiflorum and E. triplinerve. The result of the present

study shows that the plants C. alata, O. tenuiflorum and

E. triplinerve have the potential in regulation of phy-

tophagous mite species.

Proc Zool Soc (July-Dec 2011) 64(2):124–127 125

123

Discussion

Plant extract have been in use as miticides with varied

amount of success, since long. Studies on synthetic pesti-

cides revealed better results with higher mite mortality as

evident from application of dicofol, malathion and carbaryl

(0.1%) causing 100% mortality of mites within 24 h

(Krishnamoorthy 1983; Jagadish and ChannaBasavanna

1989). However, to avoid the toxic effect in environment,

botanical insecticides are being emphasized and presently

botanicals constitute 1% of the world market of insecti-

cides (Gentry 1993; Wink 1993). Extracts of Azadirachta

indica, Clerdendron inerme, Datura repens, Eucalyptus

globulus, Leucas aspera and Vitex negundo were used to

control of mite pest Tetranychus urticae and extracts of

C. inerme recorded 3.75% mortality while extracts of

E. globules recorded 13.75 and 6.25% mortality (Yathiraj

and Jagadish 1999). In the present study, mortality of

T. neocaledonicus recorded in 3% aqueous dose of plant

extracts were, 61.1% against E. triplinerve, 75% against

C. alata and 82.2% was found against O. tenuiflorum after

exposure for 72 h. On the other hand, 3% methanolic dose

of E. triplinerve and O. tenuiflorum extracts were respon-

sible for 74.4 and 93.3% mortality of mites whereas highest

mortality of 97% was recorded in 3% methanolic dose of

C. alata.

Aqueous plant extracts of Acorus calamus, Xanthium

strumarium, Polygonum hydropiper and Clerodendron in-

fortunatum lead to more than 50% mortality of red spider

mite at higher concentration (5 and 10%) under laboratory

conditions (Sarmah et al. 2009). Macchioni et al. (2004)

showed highly significant acaricidal activity of aqueous

extracts of Matricaria chamomilla against the mite

Fig. 1 Results of number of mite species killed after application of

aqueous (a) or methanolic (b) extracts of Eupatorium triplinerve(Ayapan), Cassia alata (Dadmari) and Ocimum tenuiflorum (Krish-

natulsi) in reference to the commercial available neem oil and Fungi

at the end of 72 h period. D1, D2, D3 corresponds to 1, 2 and 3%

concentration of plant extracts

Table 1 Results of ANOVA and post hoc Tukey test for comparison

of the effectiveness of the plant extracts on mortality of mite species

T. neocaledonicus

A. Between plants and solvent type used

Source Sum of squares df Mean squares F-value

Plants 13.339 2 6.670 19.96

Solvent 8.985 1 8.985 26.89

Plants 9 solvent 10.213 2 5.107 15.29

Error 106.569 318 0.334

Total 139.106 323

Post hoc Tukey test for plant species

Contrast Standardized

difference

Critical

value

Pr [ Diff

Dadmari vs. Ayapan 8.330 2.344 0.0001

Dadmari vs. Krishnatulsi 2.844 2.344 0.012

Krishnatulsi vs. Ayapan 3.043 2.344 0.007

Methanolic vs. aqueous 5.193 1.967 0.0001

B. Between plants and dose

Source Sum of squares df Mean squares F-value

Plants 13.271 2 6.636 33.395

Dose 51.966 2 25.983 130.762

Dose 9 plant 10.952 4 2.738 13.779

Error 63.188 315 0.199

Total 139.377 323

C. Between dose and solvent

Source Sum of squares df Mean squares F-value

Dose 52.139 2 26.070 117.698

Solvent 8.989 1 8.989 40.583

Dose 9 solvent 7.138 2 3.569 16.112

Error 70.436 318 0.221

Total 138.702 323

All F-values are significant at P \ 0.05 level

126 Proc Zool Soc (July-Dec 2011) 64(2):124–127

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Psoroptes cuniculi. In accordance with these findings, the

present study reveals that the methanolic and aqueous

extracts of the plants O. tenuiflorum and C. alata are

effective in regulating the population of the mites. The

results of this study appear to be promising in consideration

of the extent of regulation due to extracts of these plants as

equivalent to the commercially available biopesticides.

However, long-term studies under field conditions are

required to substantiate the potentiality of the extracts of

O. tenuiflorum and C. alata. Besides the formulations of

the plant extracts need to be evaluated since the extracts

of these plants possess antimicrobial activities (Khan et al.

2001; Rahman and Junaid 2008).

Acknowledgments The authors thank the anonymous reviewers for

the critical comments on the earlier draft of this article. The authors

are grateful to the respective Heads, Department of Zoology,

University of Calcutta, Kolkata, and The University of Burdwan,

Burdwan, India, for the facilities provided, including DST-FIST. The

authors gratefully acknowledge the help rendered by Assistant

Secretary, Ramakrishna Mission Ashram, Narendrapur, West Bengal,

India, for allowing the fieldwork in their medicinal plant garden. The

financial assistance from Ministry of Environment and Forests,

Government of India, through the Research Project (Sanction

No. 14/26/2004–ERS/RE) is thankfully acknowledged. We thank

Dr. S. K. Gupta, Senior Acarologist, for identification and valuable

suggestion during preparation of the manuscript.

References

Arnason, J.T., B.J.R. Philogene, and P. Morand. 1989. Insecticides ofplant origin. 224. ACS Symposium Series 387. Washington, DC:

American Chemical Society.

Chhillar, B.S., R. Gulati, and P. Bhatnagar. 2007. Agriculturalacarology, 355. New Delhi: Daya Publishing House.

Ehara, S., and T. Yamaguchi. 2001. Discovery of Tetranychusneocaledonicus Andre (Acari, Tetranychidae) from Amami-

Oshima Island, Japan. Plant Protection (Tokyo) 55: 268–272.

(in Japanese).

Gentry, A. H. 1993. Tropical forests diversity and the potential for

new medicinal plants. In Human medicinal agents from plants,

eds. A.D. Kinghorn, and M. F. Balandrin, 13–24. ACS Sympo-

sium Series 534, Washington DC: American Chemical Society.

Gupta, S.K. 2005. Insects and mites infesting medicinal plants inIndia, 214. Narendrapur: Kolkata: Ramakrishna Mission

Ashrama.

Helle, W., and W.P.J. Overmeer. 1985. Rearing techniques. In Spidermites. Their biology, natural enemies and control. world croppest, vol. 1A, eds. W. Helle, and M.W. Sabelis, 331–335.

Amsterdam: Elsevier.

Ismail, M.S.M., M.F.M. Soliman, M.H. El Naggar, and

M.M. Ghallab. 2007. Acaricidal activity of spinosad and

abamectin against two-spotted spider mites. Experimental andApplied Acarology 43: 129–135.

Jagadish, P.S., and G.P. ChannaBasavanna. 1989. Toxicity of pesti-

cides on Amblyseius tetranychivorous (Acari: Phytoseiidae) an

effective predator of Tetranychus ludeni (Acari:Tetranychidae).

In Progress in Acarology, vol. 2, eds. G.P. ChannaBasavanna,

and C.A. Viraktamath, 449–452. New Delhi: Oxford and IBH

Publishing.

Jbilou, R., A. Ennabili, and F. Sayah. 2006. Insecticidal activity of four

medicinal plant extracts against Tribolium castaneum (Herbst)

(Coleoptera: Tenebrionidae). African Journal of Biotechnology 5:

936–940.

Khan, M.R., M. Kihara, and A.D. Omoloso. 2001. Antimicrobial

activity of Cassia alata. Fitoterapia 72: 561–564.

Krishnamoorthy, A. 1983. Effect of some pesticides on the predatory

mite Amblyseius tetranychivorus (Gupta) (Acarina: Phytoseii-

dae). Entomon 8: 229–234.

Macchioni, F., S. Perrucci, F. Cecchi, P.L. Cioni, I. Morelli, and S.

Pampiglione. 2004. Acaricidal activity of aqueous extracts of

chamomile flowers, Matricaria chamomillai, against the mite

Psoroptes cuniculi. Medical and Veterinary Entomology 18:

205–207.

Rahman, M.S., and M. Junaid. 2008. Antimicrobial activity of leaf

extracts of Eupatorium triplinerve Vehl. against some human

pathogenic bacteria and phytopathogenic fungi. BangladeshJournal of Botany 37(1): 89–92.

Rahman, A., M. Sarmah, A.K. Phukan, M. Borthakur, and G.

Gurusubramanian. 2005. A plant having insecticidal property forthe management of tea pests. In Proceedings of 2005 interna-tional symposium on innovation in tea science and sustainabledevelopment in tea industry, 11–15 November, 2005, 731–748.

Hangzhou: Chinese Academy of Agricultural Science.

Sadana, G.L. 1985. Plant feeding mites of India, 99. New Delhi:

Kalyani Publishers.

Sarmah, M., A. Basit, and L.K. Hazarika. 1999. Effect of Polygonumhydropiper L. and Lantana camara L. on tea red spider mite.

Oligonychus coffeae. Two and a Bud 46: 20–22.

Sarmah, M., A. Rahman, A.K. Phukan, and G. Gurusubramanian.

2009. Effect of aqueous plant extracts on tea red spider mite,

Oligonychus coffeae, Neitner (Tetranychidae: Acarina) and

Stethorus gilvifrons Mulsant. African Journal of Biotechnology8: 417–423.

Sundaram, K.M.S., R. Campbell, L. Sloane, and J. Studens. 1995.

Uptake, translocation persistence and fate of azadirachtin in

aspen plants (Populustremuloides Michx.) and its effect on

pestiferous two-spotted spider mite (Tetranychus urticae Koch).

Crop Protection 14: 415–421.

Venzon, M., M.C. Rosado, A.J. Molina-Rugama, V.S. Duarte, R.

Dias, and A. Pallini. 2008. Acaricidal efficacy of neem against

Polyphagotarsonemus latus (Banks) (Acari: Tarsonemidae).

Crop Protection 27: 869–872.

Wink, M. 1993 Production and application of phytochemicals from an

agricultural perspective. In Phytochemistry and agriculture,

proceedings of phytochemical society of Europe Vol. 34 eds.

T.A. Van Beek, and H. Breteler, 171–213. Oxford: Oxford

University Press.

Yathiraj, B.R., and P.S. Jagadish. 1999. Plant extracts—future

promising tools in the integrated management of spider mite,

Tetranychus urticae (Acari: Tetranychidae). Journal of Acarol-ogy 15: 40–43.

Zar, J.H. 1999. Biostatistical Analysis, 4th ed, 663. New Delhi:

Pearson Education (Singapore) Pvt. Ltd. (Indian Branch).

Proc Zool Soc (July-Dec 2011) 64(2):124–127 127

123