Research Article

Enhanced Acaricidal Activity of Senna auriculata and Ocimum tenuiflorum Extracts with the Addition of Adjuvant against Red Spider Mite, Oligonychus coffeae Nietner (Acarina: Tetranychidae) Infesting Tea  

Pandian Kumar1 , Chandran Sundaravadivelan1 , Duraikkannu Vasantha Kumar1 , Azariah Babu2
1 Entomology Division, UPASI Tea Research Foundation, Tea Research Institute, Nirar Dam BPO, Valparai 642 127, Tamil Nadu, India
2 Tea Research Association, North Bengal Regional R&D Centre, Nagrakata 735 225, West Bengal, India
Author    Correspondence author
Journal of Tea Science Research, 2016, Vol. 6, No. 10   doi: 10.5376/jtsr.2016.06.0010
Received: 29 Nov., 2016    Accepted: 29 Nov., 2016    Published: 27 Dec., 2016
© 2016 BioPublisher Publishing Platform
Preferred citation for this article:

Kumar P., Sundaravadivelan C., Vasantha Kumar D., and Babu A., 2016, Enhanced acaricidal activity of Senna auriculata and Ocimum tenuiflorum extracts with the addition of adjuvant against red spider mite, Oligonychus coffeae nietner (Acarina : Tetranychidae) infesting tea, Journal of Tea Science Research, 6(10): 1-10 (doi: 10.5376/jtsr.2016.06.0010)


Tea (Camellia sinensis) is one of the most consumed non-alcoholic beverages in the world and has been affected by many pests which damage its quality and production globally. In this paper an attempt has been made to evaluate the aqueous leaf extracts of Senna auriculata and Ocimum tenuiflorum with and without adjuvant for their effect on red spider mite (RSM), Oligonychus coffeae (Nietner) infesting tea. Resulting, Senna auriculata, leaf extracts with adjuvant caused 90 % (LC50:1.21 and LC90:2.33 %) mortality which was greater compared to Ocimum tenuiflorum whereas 73 % (LC50:1.40 and LC90:2.81 %) was observed at 5 % concentration after 24 h exposure and very lowest mortality percent were observed both extracts without adjuvant. Ocimum tenuiflorum leaf extracts with adjuvant (0.05 %) showed highest ovicidal activity and effective ovipositional deterrence compared with S. auriculata treatment. The evaluated plant extracts with adjuvant are effective for controlling RSM and it could be an alternative, easily accessible, and eco-friendly approach for control of mite infesting tea.

Senna auriculata; Ocimum tenuiflorum; Plant extracts; Adjuvant; Red spider mite


Tea, Camellia sinensis (L) O.Kuntze belongs to family Theaceae, is a major cash crop worldwide including India. Tea leaves are known to contain active ingredients such as polyphenols (catechins and flavonoides), alkaloids (caffeine, theobromine, and theophylline), volatile oils, polysaccharides, amino acids, lipids, vitamins, inorganic elements (aluminium, fluorine and manganese) (Sharangi, 2009), and caffeine, flavonols, oleic glyceride, and linoleic glyceride (Haseqawa et al., 2011). These active metabolites having many biological activities like anti-cancer activity, lipid lowering activity, anti-carcinogenic activity, neuromuscular-blocking action, immunomodulatory effect, DNA effect, anti-viral activity, anti-bacterial activity, anti-spasmodic activity, anti-cataract activity, anti-oxidant activity, anti-diabetic activity, anti-genotoxic effect, antibacterial effect in intestine, anti-inflammatory activity, effect on oxidative stress, and chemoprotective activity (Bhatt et al., 2010). Being a monoculture crop, it is attacked by an array of insect and mite pests. Among them, red spider mite (RSM), Oligonychus coffeae (Acarina: Tetranychidae) is one of the most important arthropod pests of tea, widely distributed in India, Bangladesh, Sri Lanka, Taiwan, Burundi, Kenya, Malawi, Uganda, and Zimbabwe and is an important pest causing considerable crop loss in southern India (Muraleedharan et al., 2005; Babu et al., 2008). Infestation by this mite starts along the midrib and veins, and gradually spreads to the entire upper surface of leaves. As a result of feeding, the maintenance foliage turns ruddy bronze, making RSM infested fields conspicuous even from a distance (Figure 1 and 2) and severe infestation leads to defoliation. Some insects and mite are considered as serious pest by causing severe damages in crops which causing approximately US $500 million to $1 billion yield losses(Hazarika et al., 2009).


Chemical-based integrated control measures have been suggested for the control of RSM (Babu and Muraleedharan, 2010; Roy et al., 2012). The synthetic pesticides such as dicofol, propargite, fenpyroximate, and hexythiazox are being used for control of RSM. Recently, field evaluation of bifenazate (acramite 50 wp) for control of tea mites has been reported (Kumari et al., 2012). The using synthetic chemicals have given many profits and convenience to mankind, but a lot of them have revealed serious environmental problems and threatening to human life and cattle (Kim et al., 2005; Fetoh and Al-Shammery, 2011). Similarly, indiscriminate use of these chemicals and improper execution of pest control measures have led to various problems, such as a resurgence of primary pests (Hazarika et al., 2009), destruction of natural enemies (Mori & Gotoh, 2001; Kumral et al., 2011), development of resistance (Moghadam et al., 2012), undesirable residues on prepared tea (Chaudhuri, 1999), and contamination of environment (Gulati et al., 2010). In addition, currently using acaricides are containing organophosphates or synthetic pyrethroids and some tick species have been become resistant to these acaricides as a result of repeated exposure (Foil et al., 2004; Jayaseelan and Abdul Rahuman, 2012).



Figure 1 Healthy tea, Camellia sinensis plantation



Figure 2 Red Spider mite, Oligonychus coffeae infested tea field


To overcome the current crisis being faced by the tea industry, there is need to switch over to alternatives such as the use of plants and microbial mediated extracts (Sarmah et al., 2009), and numerous plant species possessing potential pest-controlling properties under laboratory conditions and in field also. The use of plant-based biopesticides is considered by many researchers (Roy et al., 2012; Vasanthakumar et al., 2012) for the control of many crop pests. The most important bioactive constituents of plants are steroids, terpenoids, carotenoids, flavonoids, alkaloids, tannins and glycosides. Plant extracts contains these secondary metabolites which acts the crucial role for the control of insect pest, it has been elaborately reviewed as the usage of botanicals in pest management (Isman, 2006). The crude aqueous extracts of plants contain rich source of bioactive compounds like secondary metabolites at various constitutional level, which naturally avail nontoxic product and have no degradable problem for safest environment and act as alternative source for control of insect pest (Amer and Mehlhorn, 2006; Rahuman and Venkatesan, 2008; Rahuman et al., 2009).


Previously, effect of crude phyto-extracts for the control of crop pest has been reported by many researchers. In this respect, larvicidal activity of extracts and triterpenoids from Lantana camara against Spodoptera littoralis (Lepidoptera: Noctuidae), Clavigralla tomentosicollis (Hemiptera: Coreidae) and Aphis craccivora (Homoptera: Aphididae) has been reported (Fatope et al., 2002). Campli et al. (2012) found and reported the activity of tea tree (Australian native plant, Melaleuca alternifolia) oil and nerolidol (3,7,11-trimethyl-1,6,10-dodecatrien-3-ol) against head lice and its eggs. Adulticidal and repellent properties of Cassia tora against Culex quinquefasciatus, Aedes aegypti, and Anopheles stephensi have been observed by Amerasan et al. (2012). In addition, in vitro anti-plasmodial activity of selected phytochemicals like Aglafolin, Rocaglamid, Artemisinin, Eurycoma, Quinine, and Mefloquine against Plasmodium falciparum has been reported (Astelbauer et al., 2012).


Senna auriculata (L.) Roxb. Syn. Cassia auriculata L. belongs to the family Fabaceae, and commonly found in tropical and sub-tropical regions. It is commonly called as Tanners cassia in English and, Avaram in Tamil (Figure 3). The shrub is famous for its attractive yellow flowers and the presence of active phytochemicals such as terpenoids, tannins, flavonoids, saponin, cardiac glycosides and steroids having medicinal properties (Maneemegalai and Naveen, 2010). Traditionally, plant parts leaf, flower, seed and root are used for different diseases like diabetes and anthelminthic, astringent, problems related to eye and antihyperlipidaemic effect (Pari and Latha, 2002), and skin disorders, body odour, anti-mutagenic and anti-bacterial activity (Shiradkar et al., 2011). It is also used for the treatment of ulcers, leprosy and liver disease.


Scientific classification



Figure 3 Senna auriculata (L.) Roxb. Syn. Cassia auriculata L.


Ocimum tenuiflorum L. Syn. Ocimum sanctum Linn. belonging to family Lamiaceae is also known as Tulsi in Tamil and is an erect, much branched sub-shrub 30-60 cm tall, with simple opposite green or purple leaves (Figure 4). It is cultivated for religious and medicinal purposes, specifically for its essential oil (Pattanayak et al., 2010). Ocimum sanctum contains highly complex nature of chemical composition and also having many nutrients and other biologically active compounds. The leaf volatile oil contains eugenol (1-hydroxy-2-methoxy-4-allylbenzene), eugenic acid, urosolic acid carvacrol (5-isopropyl-2-methylphenol), linalool (3,7-dimethylocta-1,6-dien-3-ol), limatrol, caryophyllene, and methyl carvicol and seed volatile oil contains fatty acids and sitosterol (Kelm et al., 2000; Shishodia et al., 2003). The leaves and stems are known to be presence of variety of active constituents like saponins, flavonoids, triterpenoids, and tannins (Jaggi et al., 2003), which may have anti-oxidant and anti-inflammatory activity. Pattanayak et al. (2010) reviewed and reported that the biologically activities such as anti-diabetic, cardiac activity, wound healing activity, radio-protective effect, genotoxicity, anti-oxidant, hypolipidemic, anti-microbial, gastroprotective, immunomodulatory effect, anti-nociceptive, anthelmintic activity, anti-inflammatory, anti-cancer, and thyroid activity of O. sanctum extracts.


Scientific classification



Figure 4 Ocimum tenuiflorum L. Syn. Ocimum sanctum Linn.


Adjuvants are used with pesticides in order to enhance the efficacy of pesticides such as herbicides, insecticides, and fungicides for control or eliminate unwanted pests. This additive can modify some property of the spray solution and improves the ability of the pesticide. Due to non-availability of the reports related to plant extracts with adjuvant against the target pest, the present study as first report was focused to explore the use of aqueous leaf extracts of the two commonly available botanicals such as O. tenuiflorum and S. auriculata alone and with adjuvant for the management of red spider mite, O. coffeae infesting tea.


2 Materials and Methods

2.1 Insect maintenance

Red spider mite, Oligonychus coffeae was collected from tea fields of UPASI Tea Experimental Farm, Valparai, Tamil Nadu, India. After field collection, spider mites were immediately transferred onto 1-year-old potted tea plants kept in a greenhouse at 25 ± 1°C and 75 ± 5% relative humidity and used as stock cultures. From the stock, adult RSM were transferred onto fresh tea leaflets (6 cm2) placed on moistened cotton pads (c.1.5 cm thick) in plastic trays (42 × 30 × 6.5 cm). Rearing trays were kept under controlled conditions at 25 ± 1°C, 75 ± 5% relative humidity, and 16 : 8 h light : dark photoperiod. Withered and drying leaves were regularly replaced.


2.2 Preparation of aqueous leaf extracts

Fresh and healthy, leaves of Senna auriculata and Ocimum tenuiflorum were collected from Viluppanoor, Tamil Nadu, India. The freshly harvested plant leaves were washed thoroughly in tap water, rinsed well in distilled water, and shade-dried at room temperature (35±1 °C). These dried leaves were powdered mechanically using blending machine and the powder was used for preparation of aqueous leaf extracts. Aqueous leaves extracts were prepared by mixing 3, 4 and 5 g of dried leaves powder in 100 ml of double distilled water. This mixture suspension was mixed well and left for 5 h without disturbance, then filtered through Whatman filter paper (No.1). The filtrate was used to find the adulticidal, ovicidal and Ovipositional deterrence activity against the target pest red spider mite, O. coffeae a major pest in tea.


2.3 Adjuvants

For enhance the bio-efficacy, Ig Surf 2115, as adjuvant (0.05%) was mixed while soaking the dried leaves powder of S. auriculata and O. tenuiflorum in double distilled water before the extraction and simultaneously, extracts without adjuvant were also prepared for the comparison study.


2.4 Adulticidal activity

Leaf discs of 20-mm diameter cut from mature tea leaves were placed in Petri dishes lined with water-saturated cotton wool. Ten adult female mites were introduced onto the surface of the discs using a fine camel-hair brush under a binocular microscope (Olympus No. 1220). Direct spraying technique (topical bioassay) was used to evaluate the efficacy of leaf extracts on adult mites. Desired concentrations of leaf extracts (3, 4, and 5% with and without adjuvant 0.05%) were prepared using distilled water. Each leaf discs inside the petri dishes was sprayed with the same amount of leaf extracts using a manual glass atomizer (VENSIL 50 ml). Leaf discs sprayed with distilled water were kept as control. The surviving mites were counted after 24, 48, 72, and 96 h exposure and the mean mortality rate was calculated. Each experiment was replicated five times.


2.5 Ovicidal activity

To provide eggs for this bioassay, leaf discs of 20-mm diameter were placed in Petri dishes lined with water-saturated cotton wool. Five gravid female mites were introduced onto the surface of the discs 24 h before the start of the experiment and allowed to lay eggs. Then the adults were removed and the number of eggs was adjusted to 10 eggs/ leaf disc. The leaf discs containing the eggs were sprayed with different concentrations of leaf extracts (3, 4, and 5% with and without adjuvant 0.05%). Leaf discs sprayed with distilled water were kept as control. The discs were dried for at least 30 min. After drying, the discs were placed at 27 ± 2°C and 65 ± 5% relative humidity. All discs were examined on a daily basis for eight successive days. Egg hatching rate and development of young mites were determined by counting the numbers of eggs hatched and of surviving larvae or nymphs on the discs using a binocular stereomicroscope (Olympus No. 1220). Five replicates were maintained for each treatment.


2.6 Ovipositional deterrence activity

Ovipositional deterrence was evaluated by allowing the adult females to lay eggs on leaf discs of 20-mm diameter treated with the different concentrations of leaf extracts (3, 4, and 5% with and without adjuvant 0.05%). Females were released onto the treated leaf discs as well as onto control (water-treated) leaf discs for egg laying. The degree of deterrence was assessed in terms of variations in the number of eggs laid by the females on the untreated and treated leaf discs every 24 h for 4 days.


2.7 Statistical analysis

Data on the mortality rate of eggs, adults and data on ovipositional deterrence were subjected to analysis of variance and means were separated by Tukey’s test. The discrimination quotient (DQ) was calculated using the following formula:

Note: C is the number of eggs on control leaves and T is the number of eggs on treated leaves.


The lethal concentrations, 50 % and 90 % (LC50 and LC90) were determined and slopes, lower and upper confidence level were calculated using statistical tool SPSS 13.0 ver.


3 Results and Discussion

3.1 Enhanced acaricidal activity

The introduction of more effective biocontrol methods will reduce the rates of chemical pesticides and prevent, or at least delay the development of resistance in target pests. Based on this concept, in the present study the results revealed that the enhancing acaricidal activity of aqueous leaf extracts of S. auriculata and O. tenuiflorum with and without adjuvant against egg, ovipositional and adult of red spider mite, O. coffeae a major pest in tea. The data on the adulticidal activity of aqueous leaf extracts of S. auriculata and O. tenuiflorum with and without adjuvant are presented in the table (Table 1). Aqueous leaf extracts of S. auriculata with adjuvant caused 90% mortality at 5% (0.05% adjuvant) concentration after 24 h exposure which was compared to aqueous leaf extracts without adjuvant caused 70% mortality. Their lethal concentration, LC50 and LC90 for leaf extracts with adjuvant 0.01 and 2.33% and without adjuvant 1.93 and 3.28% were observed respectively (Table 2). Both, leaf extracts with and without adjuvant caused 100% mortality after 72 h exposure. Aqueous extracts of Acorus calamus, Xanthium strumarium, Polygonum hydropiper and Clerodendron infortunatum showed effective mortality at 2.5, 5.0 and 10.0% (w/v) tested concentrations on tea red spider mite, Oligonychus coffeae (Sarmah et al., 2009) and similarly, Ambrosia maritimal ethanolic extracts caused 93.33% mortality while compared with Duranta plumeria (69%), and Cuminum cyminum (64.67%) in adult females of the date palm dust mite, Oligonychus afrasiaticus (Fetoh and Al-Shammery, 2011).



Table 1 Acaricidal activity of S. auriculata and O. tenuiflorum leaf extracts with adjuvant against red spider mite, O. coffeae

Note: aValues represent mean of five replicates: Mean±SE followed by the same letter do not differ significantly at (P=0.05) according to Tukey’s test.



Table 2 The LC50 and LC90 of effect of O. santum and C. auriculata extracts on adult red spider mite, O. coffeae


Ocimum tenuiflorum leaf extracts with adjuvant caused 100% (LC50: 1.40; LC90: 2.81%) mortality, and without adjuvant showed 48% (LC50: 4.82; LC90: 9.52%) after 72 h exposure. Likewise, acaricidal activity of Xanthium strumarium, Acorus calamus, Polygonum hydropiper and Clerodendron infortunatum extracts were observed against Oligonychus coffeae and they caused 91.8, 88.7, 84.2, and 100% mortality at 10% concentration after 72 h exposure (Sarmah et al., 2009). Previously, many researchers have been reported the potential activity of plant crude extracts against various crop pest and insect vectors. In this respect, in vitro adulticidal activity of Chenopodium album, Chrysanthemum cinerariifolium, Bauhinia variegata, Cuscuda reflexa, Alianthus excelsa, Calotropis gigantea, and Annona squamosa extracts has been observed against Haemonchus contortus (Yadav et al., 2010). Recently, 100% larvicidal activity was observed on fourth instar larvae of American boll warm, Helicoverpa armigera treated with Calotropis procera and Argimone mexicana leaf extracts at 100 ppm concentration (Lall et al., 2013). The observed results indicated that aqueous leaf extracts of O. tenuiflorum and S. auriculata with adjuvant showed highly potent against O. coffeae compared with leaf extracts without adjuvant and no mortality was observed in control.


3.2 Ovicidal activity

The ovicidal activity of S. auriculata and O. tenuiflorum aqueous leaf extracts with and without adjuvant was observed (Table 3) at 3, 4, and 5% tested concentration. Leaf extracts of S. auriculata with adjuvant showed 68% egg mortality than that of without adjuvant treatment, whereas, 58 % was observed. Similarly, 74 and 60% egg mortality were observed treated with O. tenuiflorum leaf extracts with and without adjuvant at 5% concentration, respectively. The toxic nature of leaf extracts to the eggs of red spider mites have been reported earlier (Sarmah et al., 2009; Vasanthakumar et al., 2012). Prabhu et al. (2012) observed that aqueous leaf extracts of Calotropis gigantea caused 100% egg mortality on Helicoverpa armigera at 10% concentration and 77.66% inhibition of egg hatchability was recorded while tested on 24 h old eggs of Spodoptera litura (Manikantan, 2003). In treatment, the increased egg mortality was observed in O. tenuiflorum leaf extracts with adjuvant compared to S. auriculata leaf extracts with adjuvant while treated against O. coffeae eggs and no egg mortality was observed in control. The chemical substance present in the botanical products blocks the micropyle region of the egg, thereby preventing gaseous exchange and ultimately killing the embryo during the egg stage (Sarmah et al., 2009).



Table 3 Ovicidal activity of aqueous leaf extracts against red spider mite

Note: *Values represent mean of five replicates: Mean ± SE followed by the same letter do not differ significantly at (P=0.05) according to Tukey’s test.


3.3 Ovipositional deterrence activity

In addition to adulticidal and ovicidal activity, the tested plant extracts with and without adjuvant also exerted ovipositional deterrence in RSM by preventing them from laying eggs on the treated leaf discs. Ovipositional deterrence and DQ value of S. auriculata and O. tenuiflorum leaf extracts with and without adjuvant are presented in the table (Table 4). The DQ, which has a range from 0 to 1, is an index for determination of the effect of any chemical on the ovipositional behavior of insects. When the leaves sprayed with different concentration of leaf extracts with and without adjuvant were provided for egg laying, adult mites showed some discrimination between the treated and control leaves with a variation in the number of eggs laid. In the present study, the higher DQ value of 0.77 was observed in the treatment of O. tenuiflorum leaf extracts with adjuvant and very low 0.28 was observed in S. auriculata leaf extracts without adjuvant treatment at 5% concentration. Increased numbers of laid eggs were observed in control. Our findings are in line with Vasanthakumar et al. (2012) who have already documented the ovipositional deterrent nature in RSM when treated with leaf extracts of Vitex negundo, Gliricidia maculata, Wedelia chinensis, Morinda tinctoria and Pongamia glabra. Recently, Mishra et al. (2014) reported that leaves of Aegle marmelos, Mentha arvensis, peels of Citrus reticulate, and clove of Syzygium aromaticum mediated essential oils significantly reduced the oviposition capacity on rice weevil, Sitophilus oryzae. In addition, aqueous extracts of leaf of Artemisia nilagirica, Cassia auriculata, Cassia siamia, Percularia daemia, root of Acorus calamus, and peel of Citrus aurantium significantly reduced the oviposition to 19.25, 30.92, 84.66, 47.61, 32.61, and 82.11% respectively, at higher concentration of 10% (Jayakumar, 2010).



Table 4 Ovipositional deterrence and Discrimination Quotient (DQ) of plant extracts against red spider mite

Note: aValues represent mean of five replicates: Mean ± SE followed by the same letter do not differ significantly at P = 0.05 according to Tukey’s test.


3.4 Possible mechanism

The plant extracts contain active metabolites could be converted to toxic metabolites by enzymes in the body. Primarily, it may affect insects nerves and inhibit their energy metabolism. In this respect, the presence of monoterpenoids in Aegle marmelos essential oil has been responsible to inhibit the reproduction of stored insects at several steps of their life cycles (Mishra et al., 2014). Specifically, the reduction in oviposition and adult emergence may be due to the suffocation and inhibition of various biosynthesis processes of the insects at developmental stages. The physiological changes due to metabolic disruption by applying the pesticides during insect development may finally leads to the formation of abnormal adult or death.

4 Conclusion

Our findings have shown that the tested S. auriculata and O. tenuiflorum leaf extracts possesses adulticidal, ovicidal and ovipositional deterrent activity against red spider mite. Moreover, addition of adjuvant with leaf extracts enhanced the efficacy in controlling red spider mites. Beyond this first report, further studies are needed under field conditions for the effective use of these leaf extracts alone with adjuvant in developing environmentally safe method to manage red spider mite, O. coffeae infesting tea.



The authors are grateful to Dr. N. Muraleedharan, Advisor, and Dr. P. Mohan Kumar, Director, UPASI Tea Research Institute, Tea Research Foundation, for providing the facilities and constant encouragement.



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Journal of Tea Science Research
• Volume 6
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